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Sample records for soluble globular proteins

  1. Hydrophobicity diversity in globular and nonglobular proteins measured with the Gini index.

    Science.gov (United States)

    Carugo, Oliviero

    2017-12-01

    Amino acids and their properties are variably distributed in proteins and different compositions determine all protein features, ranging from solubility to stability and functionality. Gini index, a tool to estimate distribution uniformity, is widely used in macroeconomics and has numerous statistical applications. Here, Gini index is used to analyze the distribution of hydrophobicity in proteins and to compare hydrophobicity distribution in globular and intrinsically disordered proteins. Based on the analysis of carefully selected high-quality data sets of proteins extracted from the Protein Data Bank (http://www.rcsb.org) and from the DisProt database (http://www.disprot.org/), it is observed that hydrophobicity is distributed in a more diverse way in intrinsically disordered proteins than in folded and soluble globular proteins. This correlates with the observation that the amino acid composition deviates from the uniformity (estimate with the Shannon and the Gini-Simpson indices) more in intrinsically disordered proteins than in globular and soluble proteins. Although statistical tools tike the Gini index have received little attention in molecular biology, these results show that they allow one to estimate sequence diversity and that they are useful to delineate trends that can hardly be described, otherwise, in simple and concise ways. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  2. Modular organization of proteins containing C1q-like globular domain.

    Science.gov (United States)

    Kishore, U; Reid, K B

    1999-05-01

    The first step in the activation of the classical pathway of complement cascade by immune complexes involves the binding of the six globular heads of C1q to the Fc regions of immunoglobulin G (IgG) or immunoglobulin M (IgM). The globular heads of C1q are located C-terminal to the six triple-helical stalks present in the molecule, each head is considered to be composed of the C-terminal halves (3 x 135 residues) of one A-, one B- and one C-chain. It is not known if the C-terminal globular regions, present in each of the three types of chains, are independently folded modules (with each chain having distinct binding properties towards immunoglobulins) or whether the different binding functions of C1q are dependent upon a globular structure which relies on contributions from all three chains. Recent reports of recombinant production and characterisation of soluble globular head regions of all the three chains indicate that the globular regions of C1q may adopt a modular organization, i.e., each globular head of C1q may be composed of three, structurally and functionally, independent domains, thus retaining multivalency in the form of a heterotrimer. Modules of the same type as the C1q C-terminal module are also found in a variety of noncomplement proteins that include the C-terminal regions of the human type VIII and type X collagens, precerebellin, the chipmunk hibernation proteins, the human endothelial cell protein, multimerin, the serum protein, Acrp-30 which is secreted from mouse adipocytes, and the sunfish inner-ear specific structural protein. The C1q molecule is the only one of these proteins for which, to date, a function has been ascribed to the module. The existence of a shared structural region between C1q and certain collagens may suggest an evolutionarily common ancestral precursor. Various structural and biochemical data suggest that these modules may be responsible for multimerisation through patches of aromatic residues within them.

  3. Cold gelation of globular proteins

    NARCIS (Netherlands)

    Alting, A.C.

    2003-01-01

    Keywords : globular proteins, whey protein, ovalbumin, cold gelation, disulfide bonds, texture, gel hardnessProtein gelation in food products is important to obtain desirable sensory and textural properties. Cold gelation is a novel method to produce protein-based gels. It is a two step process in

  4. Interactions between globular proteins and F-actin in isotonic saline solution.

    Science.gov (United States)

    Lakatos, S; Minton, A P

    1991-10-05

    Solutions of each of three different globular proteins (cytochrome c, chromophorically labeled serum albumin, and chromophorically labeled aldolase), mixed with another unlabeled globular protein or with fibrous actin, were prepared in pH 8.0 Tris-HCl buffer containing 0.15 M NaCl. Each solution was centrifuged at low speed, at 5 degrees C, until unassociated globular protein in solution achieved sedimentation equilibrium. Individual absorbance gradients of both macrosolutes in the mixtures subsequent to centrifugation were obtained via optical scans of the centrifuge tubes at two wavelengths. The gradients of each macrosolute in mixtures of two globular proteins revealed no association of globular proteins under the conditions of these experiments, but perturbation of the gradients of serum albumin, aldolase, and cytochrome c in the presence of F-actin indicated association of all three globular proteins with F-actin. Perturbation of actin gradients in the presence of serum albumin and aldolase suggested partial depolymerization of the F-actin by the globular protein. Analysis of the data with a simple phenomenological model relating free globular protein, bound globular protein, and total actin concentration provided estimates of the respective equilibrium constants for association of serum albumin and aldolase with F-actin, under the conditions of these experiments, of the order of 0.1 microM-1.

  5. Nanofibers made of globular proteins.

    Science.gov (United States)

    Dror, Yael; Ziv, Tamar; Makarov, Vadim; Wolf, Hila; Admon, Arie; Zussman, Eyal

    2008-10-01

    Strong nanofibers composed entirely of a model globular protein, namely, bovine serum albumin (BSA), were produced by electrospinning directly from a BSA solution without the use of chemical cross-linkers. Control of the spinnability and the mechanical properties of the produced nanofibers was achieved by manipulating the protein conformation, protein aggregation, and intra/intermolecular disulfide bonds exchange. In this manner, a low-viscosity globular protein solution could be modified into a polymer-like spinnable solution and easily spun into fibers whose mechanical properties were as good as those of natural fibers made of fibrous protein. We demonstrate here that newly formed disulfide bonds (intra/intermolecular) have a dominant role in both the formation of the nanofibers and in providing them with superior mechanical properties. Our approach to engineer proteins into biocompatible fibrous structures may be used in a wide range of biomedical applications such as suturing, wound dressing, and wound closure.

  6. Statistical interior properties of globular proteins

    International Nuclear Information System (INIS)

    Zhou-Ting, Jiang; Tai-Quan, Wu; Lin-Xi, Zhang; Ting-Ting, Sun

    2009-01-01

    The character of forming long-range contacts affects the three-dimensional structure of globular proteins deeply. As the different ability to form long-range contacts between 20 types of amino acids and 4 categories of globular proteins, the statistical properties are thoroughly discussed in this paper. Two parameters N C and N D are defined to confine the valid residues in detail. The relationship between hydrophobicity scales and valid residue percentage of each amino acid is given in the present work and the linear functions are shown in our statistical results. It is concluded that the hydrophobicity scale defined by chemical derivatives of the amino acids and nonpolar phase of large unilamellar vesicle membranes is the most effective technique to characterise the hydrophobic behavior of amino acid residues. Meanwhile, residue percentage P i and sequential residue length L i of a certain protein i are calculated under different conditions. The statistical results show that the average value of P i as well as L i of all-α proteins has a minimum among these 4 classes of globular proteins, indicating that all-α proteins are hardly capable of forming long-range contacts one by one along their linear amino acid sequences. All-β proteins have a higher tendency to construct long-range contacts along their primary sequences related to the secondary configurations, i.e. parallel and anti-parallel configurations of β sheets. The investigation of the interior properties of globular proteins give us the connection between the three-dimensional structure and its primary sequence data or secondary configurations, and help us to understand the structure of protein and its folding process well. (cross-disciplinary physics and related areas of science and technology)

  7. Production of soluble mammalian proteins in Escherichia coli: identification of protein features that correlate with successful expression

    Directory of Open Access Journals (Sweden)

    Perera Rajika L

    2004-12-01

    Full Text Available Abstract Background In the search for generic expression strategies for mammalian protein families several bacterial expression vectors were examined for their ability to promote high yields of soluble protein. Proteins studied included cell surface receptors (Ephrins and Eph receptors, CD44, kinases (EGFR-cytoplasmic domain, CDK2 and 4, proteases (MMP1, CASP2, signal transduction proteins (GRB2, RAF1, HRAS and transcription factors (GATA2, Fli1, Trp53, Mdm2, JUN, FOS, MAD, MAX. Over 400 experiments were performed where expression of 30 full-length proteins and protein domains were evaluated with 6 different N-terminal and 8 C-terminal fusion partners. Expression of an additional set of 95 mammalian proteins was also performed to test the conclusions of this study. Results Several protein features correlated with soluble protein expression yield including molecular weight and the number of contiguous hydrophobic residues and low complexity regions. There was no relationship between successful expression and protein pI, grand average of hydropathicity (GRAVY, or sub-cellular location. Only small globular cytoplasmic proteins with an average molecular weight of 23 kDa did not require a solubility enhancing tag for high level soluble expression. Thioredoxin (Trx and maltose binding protein (MBP were the best N-terminal protein fusions to promote soluble expression, but MBP was most effective as a C-terminal fusion. 63 of 95 mammalian proteins expressed at soluble levels of greater than 1 mg/l as N-terminal H10-MBP fusions and those that failed possessed, on average, a higher molecular weight and greater number of contiguous hydrophobic amino acids and low complexity regions. Conclusions By analysis of the protein features identified here, this study will help predict which mammalian proteins and domains can be successfully expressed in E. coli as soluble product and also which are best targeted for a eukaryotic expression system. In some cases

  8. Pyroelectricity in globular protein lysozyme films

    Science.gov (United States)

    Stapleton, A.; Noor, M. R.; Haq, E. U.; Silien, C.; Soulimane, T.; Tofail, S. A. M.

    2018-03-01

    Pyroelectricity is the ability of certain non-centrosymmetric materials to generate an electric charge in response to a change in temperature and finds use in a range of applications from burglar alarms to thermal imaging. Some biological materials also exhibit pyroelectricity but the examples of the effect are limited to fibrous proteins, polypeptides, and tissues and organs of animals and plants. Here, we report pyroelectricity in polycrystalline aggregate films of lysozyme, a globular protein.

  9. Microwave-enhanced folding and denaturation of globular proteins

    DEFF Research Database (Denmark)

    Bohr, Henrik; Bohr, Jakob

    2000-01-01

    It is shown that microwave irradiation can affect the kinetics of the folding process of some globular proteins, especially beta-lactoglobulin. At low temperature the folding from the cold denatured phase of the protein is enhanced, while at a higher temperature the denaturation of the protein from...... its folded state is enhanced. In the latter case, a negative temperature gradient is needed for the denaturation process, suggesting that the effects of the microwaves are nonthermal. This supports the notion that coherent topological excitations can exist in proteins. The application of microwaves...

  10. Microstructure and rheology of globular protein gels in the presence of gelatin

    NARCIS (Netherlands)

    Ersch, C.; Meinders, M/B.J.; Bouwman, W.G.; Nieuwland, M.; Linden, E. van der; Venema, P.; Martin, A.H.

    2016-01-01

    The microstructure and rheological response of globular protein gels (whey protein isolate (WPI) and soy protein isolate (SPI)) in the presence of gelatin (type A, type B and hydrolyzed type A) was investigated. Microstructural information was obtained using a combination of confocal laser scanning

  11. [Study of beta-turns in globular proteins].

    Science.gov (United States)

    Amirova, S R; Milchevskiĭ, Iu V; Filatov, I V; Esipova, N G; Tumanian, V G

    2005-01-01

    The formation of beta-turns in globular proteins has been studied by the method of molecular mechanics. Statistical method of discriminant analysis was applied to calculate energy components and sequences of oligopeptide segments, and after this prediction of I type beta-turns has been drawn. The accuracy of true positive prediction is 65%. Components of conformational energy considerably affecting beta-turn formation were delineated. There are torsional energy, energy of hydrogen bonds, and van der Waals energy.

  12. Globular conformation of some ribosomal proteins in solution

    International Nuclear Information System (INIS)

    Serdyuk, I.N.; Spirin, A.S.

    1978-01-01

    The possibility that such RNA-binding proteins of the 30 S subparticle as S4, S7, S8 and S16 exist in the form of compact globules in solution has been explored experimentally. These proteins have been studied in D 2 O solution by neutron scattering to measure their radii of gyration. This type of radiation using D 2 O as a solvent provides the maximum 'contrast', that is the maximum difference between the scattering of the protein and the solvent. It allowed measurements to be made using protein at <= 1.5 mg/ml. The radii of gyration for the ribosomal proteins S4, S7, S8 and S16 were found to be relatively small corresponding to the radii of gyration of compact globular proteins of the same molecular weights. (Auth.)

  13. Infrared and laser-Raman spectroscopic studies of thermally-induced globular protein gels.

    Science.gov (United States)

    Clark, A H; Saunderson, D H; Suggett, A

    1981-03-01

    Infrared and laser-Raman spectroscopy have been used to follow secondary structure changes during the heat-set gelation of a number of aqueous (D2O) globular protein solutions. Measurements of the infrared Amide I' absorption band around 1650 cm-1, for BSA gels of varying clarity and texture, have shown that the very considerable variations in network structure underlying these materials are not reflected in obvious differences in secondary structure. In all cases aggregation is accompanied by development of beta-sheet of a kind common in fibrous protein systems, but for BSA at least this does not appear to vary significantly in amount from one gel type to another. Infrared studies of gels formed from other protein systems have confirmed this tendency for beta-sheet to develop during aggregation, and the tendency is further substantiated by laser-Raman evidence which provides the extra information that in most of the examples studied alpha-helix content simultaneously falls. From these, and other observations, some generalisations are made about the thermally-induced sol-to-gel transformations of globular proteins.

  14. The small angle x-ray scattering of globular proteins in solution during heat denaturation

    Science.gov (United States)

    Banuelos, Jose; Urquidi, Jacob

    2008-10-01

    The ability of proteins to change their conformation in response to changes in their environment has consequences in biological processes like metabolism, chemical regulation in cells, and is believed to play a role in the onset of several neurodegenerative diseases. Factors such as a change in temperature, pressure, and the introduction of ions into the aqueous environment of a protein can give rise to the folding/unfolding of a protein. As a protein unfolds, the ratio of nonpolar to polar groups exposed to water changes, affecting a protein's thermodynamic properties. Using small angle x-ray scattering (SAXS), we are currently studying the intermediate protein conformations that arise during the folding/unfolding process as a function of temperature for five globular proteins. Trends in the observed intermediate structures of these globular proteins, along with correlations with data on protein thermodynamics may help elucidate shared characteristics between all proteins in the folding/unfolding process. Experimental design considerations will be discussed and preliminary results for some of these systems will be presented.

  15. Transiently disordered tails accelerate folding of globular proteins.

    Science.gov (United States)

    Mallik, Saurav; Ray, Tanaya; Kundu, Sudip

    2017-07-01

    Numerous biological proteins exhibit intrinsic disorder at their termini, which are associated with multifarious functional roles. Here, we show the surprising result that an increased percentage of terminal short transiently disordered regions with enhanced flexibility (TstDREF) is associated with accelerated folding rates of globular proteins. Evolutionary conservation of predicted disorder at TstDREFs and drastic alteration of folding rates upon point-mutations suggest critical regulatory role(s) of TstDREFs in shaping the folding kinetics. TstDREFs are associated with long-range intramolecular interactions and the percentage of native secondary structural elements physically contacted by TstDREFs exhibit another surprising positive correlation with folding kinetics. These results allow us to infer probable molecular mechanisms behind the TstDREF-mediated regulation of folding kinetics that challenge protein biochemists to assess by direct experimental testing. © 2017 Federation of European Biochemical Societies.

  16. Determination of soluble protein contents from RVNRL

    International Nuclear Information System (INIS)

    Wan Manshol Wan Zin; Nurulhuda Othman

    1996-01-01

    This project was carried out to determine the soluble protein contents on RVNRL film vulcanisates, with respect to the RVNRL storage time, gamma irradiation dose absorbed by the latex and the effect of different leaching time and leaching conditions. These three factors are important in the hope to determine the best possible mean of minimizing the soluble protein contents in products made from RVNRL. Within the nine months storage period employed in the study, the results show that, the longer the storage period the less the soluble protein extracted from the film samples. Gamma irradiation dose absorbed by the samples, between 5.3 kGy to 25.2 kGy seems to influence the soluble protein contents of the RVNRL films vulcanisates. The higher the dose the more was the soluble protein extracted from the film samples. At an absorbed dose of 5.3 kGy and 25.2 kGy, the soluble contents were 0. 198 mg/ml and 0.247 mg/ml respectively. At a fixed leaching temperature, the soluble proteins increases with leaching time and at a fixed leaching time, the soluble proteins increases with leaching temperature. ne highest extractable protein contents was determined at a leaching time of 10 minutes and leaching temperature of 90'C The protein analysis were done by using Modified Lowry Method

  17. A KINETIC MODEL FOR MONO-LAYER GLOBULAR PROTEIN ADSORPTION ON SOLID/LIQUID INTERFACES

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    Kamal I. M. Al-Malah

    2012-12-01

    Full Text Available A kinetic model was derived for globular protein adsorption. The model takes into account the three possible scenarios of a protein molecule in solution, being exposed to an interface: adsorption step from the solution to the interface; the possible desorption back into the solution; and the surface-induced unfolding or spreading of the protein unto the substrate surface. A globular protein molecule is visualized as a sphere with radius D. In addition to the general case of protein adsorption, which portrays either the surface coverage (Theta or surface concentration (� as a function of the adsorption time, special cases, like equilibrium condition, lowsurface coverage, irreversible, and Langmuirian were also presented and treated in light of the derived model. The general model was simplified for each of the subset cases. The irreversibility versus reversibility of protein adsorption was discussed. The substrate surface energetics or effects are accounted for via the proposition of the percent relative change in D/V ratio for the adsorbing protein, called (D/VPRC parameter. (D/VPRC is calculated with respect to the monolayer surface concentration of protein, where the latter is given by D/Vratio. This can be used as a landmark to protein adsorption isotherms or even kinetics. This is visualized as an indicator for solid substrate effects on the adsorbing proteins. (D/VPRC can be zero (fresh monolayer, negative (aged monolayer, or positive (multi-layer. The reference surface concentration is reported for some selected proteins.

  18. Strong Keratin-like Nanofibers Made of Globular Protein

    Science.gov (United States)

    Dror, Yael; Makarov, Vadim; Admon, Arie; Zussman, Eyal

    2008-03-01

    Protein fibers as elementary structural and functional elements in nature inspire the engineering of protein-based products for versatile bio-medical applications. We have recently used the electrospinning process to fabricate strong sub-micron fibers made solely of serum albumin (SA). This raises the challenges of turning a globular non-viscous protein solution into a polymer--like spinnable solution and producing keratin-like fibers enriched in inter S-S bridges. A stable spinning process was achieved by using SA solution in a rich trifluoroethanol-water mixture with β-mercaptoethanol. The breakage of the intra disulfide bridges, as identified by mass spectrometry, together with the denaturing alcohol, enabled a pronounced expansion of the protein. This in turn, affects the rheological properties of the solution. X-ray diffraction pattern of the fibers revealed equatorial orientation, indicating the alignment of structures along the fiber axis. The mechanical properties reached remarkable average values (Young's modulus of 1.6GPa, and max stress of 36MPa) as compared to other fibrous protein nanofibers. These significant results are attributed to both the alignment and inter disulfide bonds (cross linking) that were formed by spontaneous post-spinning oxidation.

  19. Dynamics of proteins at low temperatures: fibrous vs. globular

    Science.gov (United States)

    Foucat, L.; Renou, J.-P.; Tengroth, C.; Janssen, S.; Middendorf, H. D.

    We have measured quasielastic neutron scattering from H2O-hydrated collagen and haemoglobin at Tτ>10 ps. Relative to haemoglobin, the 200-K dynamic transition is shifted upward by 20-25 K in collagen, and the T-dependence of m.-sq. displacements derived from Sqe(Q;T) suggests that in triple-helical systems there are three rather than two regimes: one up to around 120K (probably purely harmonic), an intermediate quasiharmonic region with a linear dependence up to 240K, followed by a steeper nonlinear rise similar to that in globular proteins.

  20. Comparing side chain packing in soluble proteins, protein-protein interfaces, and transmembrane proteins.

    Science.gov (United States)

    Gaines, J C; Acebes, S; Virrueta, A; Butler, M; Regan, L; O'Hern, C S

    2018-05-01

    We compare side chain prediction and packing of core and non-core regions of soluble proteins, protein-protein interfaces, and transmembrane proteins. We first identified or created comparable databases of high-resolution crystal structures of these 3 protein classes. We show that the solvent-inaccessible cores of the 3 classes of proteins are equally densely packed. As a result, the side chains of core residues at protein-protein interfaces and in the membrane-exposed regions of transmembrane proteins can be predicted by the hard-sphere plus stereochemical constraint model with the same high prediction accuracies (>90%) as core residues in soluble proteins. We also find that for all 3 classes of proteins, as one moves away from the solvent-inaccessible core, the packing fraction decreases as the solvent accessibility increases. However, the side chain predictability remains high (80% within 30°) up to a relative solvent accessibility, rSASA≲0.3, for all 3 protein classes. Our results show that ≈40% of the interface regions in protein complexes are "core", that is, densely packed with side chain conformations that can be accurately predicted using the hard-sphere model. We propose packing fraction as a metric that can be used to distinguish real protein-protein interactions from designed, non-binding, decoys. Our results also show that cores of membrane proteins are the same as cores of soluble proteins. Thus, the computational methods we are developing for the analysis of the effect of hydrophobic core mutations in soluble proteins will be equally applicable to analyses of mutations in membrane proteins. © 2018 Wiley Periodicals, Inc.

  1. Co-evolutionary constraints of globular proteins correlate with their folding rates.

    Science.gov (United States)

    Mallik, Saurav; Kundu, Sudip

    2015-08-04

    Folding rates (lnkf) of globular proteins correlate with their biophysical properties, but relationship between lnkf and patterns of sequence evolution remains elusive. We introduce 'relative co-evolution order' (rCEO) as length-normalized average primary chain separation of co-evolving pairs (CEPs), which negatively correlates with lnkf. In addition to pairs in native 3D contact, indirectly connected and structurally remote CEPs probably also play critical roles in protein folding. Correlation between rCEO and lnkf is stronger in multi-state proteins than two-state proteins, contrasting the case of contact order (co), where stronger correlation is found in two-state proteins. Finally, rCEO, co and lnkf are fitted into a 3D linear correlation. Copyright © 2015 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  2. Effect of a low-frequency magnetic field on the structure of globular blood proteins

    Science.gov (United States)

    Zalesskaya, G. A.; Ulashchik, V. S.; Mit'kovskaya, N. P.; Laskina, O. V.; Kuchinskii, A. V.

    2007-09-01

    We used IR Fourier absorption spectra of blood to study changes in the structure of globular blood proteins with extracorporeal autohemomagnetotherapy, used to treat ischemic heart disease. We compare the spectra of blood before and after magnetotherapy in the regions: amide I (1655 cm-1), amide II (1545 cm-1), amide III (1230-1350 cm-1), amide IV and amide V (400-700 cm-1). We have shown that pronounced changes in the spectra in the indicated regions on direct exposure of blood in vivo to a low-frequency pulsed magnetic field are connected with conformational changes in the secondary structure of globular blood proteins, which are apparent in the increase in the contribution of the α-helix conformation. We discuss the magnetotherapy-initiated appearance of new IR absorption bands at 1018 and 1038 cm-1 and an increase in the intensity of a number of other bands located in the 1000-1200 cm-1 region, which suggests a change in the concentration of some blood components.

  3. X-radiation effect on soluble proteins of gastric mucosa

    International Nuclear Information System (INIS)

    Sukhomlinov, B.F.; Chajka, Ya.P.; Fedorovich, A.N.

    1979-01-01

    Using the method of electrophoresis in agar gel soluble proteins of gastric mucosa of rats were separated into 11 fractions. Proteins posessing a proteolytic (pH 1.8) and lipase (pH 7.4) activity were localized within the second and third prealbumin fractions. Soluble proteins of gastric mucosa contain glyco- and lipoproteid complexes. Exposure of rats to 1000 R of X-rays induces quantitative redistribution within the electrophoretic spectrum of soluble proteins and a considerable disturbance of the proteolytic activity of total soluble proteins throughout the entire period of observation (from 10 min to 72h)

  4. Characterization of soluble protein BCP 11/24 from bovine corneal epithelium, different from the principal soluble protein BCP 54

    NARCIS (Netherlands)

    Bakker, C.; Pasmans, S.; Verhagen, C.; van Haren, M.; van der Gaag, R.; Hoekzema, R.

    1992-01-01

    The water-soluble fraction of bovine corneal epithelium was analysed by polyacrylamide gel electrophoresis in the presence of SDS (SDS-PAGE). Next to the principal soluble protein BCP 54, which has recently been identified as a corneal aldehyde dehydrogenase (ALDH), another abundant protein was

  5. Omnipresence of the polyproline II helix in fibrous and globular proteins.

    Science.gov (United States)

    Esipova, Natalia G; Tumanyan, Vladimir G

    2017-02-01

    Left-handed helical conformation of a polypeptide chain (PPII) is the third type of the protein backbone structure. This conformation universally exists in fibrous, globular proteins, and biologically active peptides. It has unique physical and chemical properties determining a wide range of biological functions, from the protein folding to the tissue differentiation. New examples of the structure have been appearing in spite of difficulties in their detection and investigation. The annotation and prediction of the PPII was also a challenging task. Recently, many PPII motifs with new and/or unexpected functions are being accumulated in databases. In this review we describe the major structural and dynamic forms of PPII, the diversity of its functions, and the role in different biological processes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Relationship between Molecular Structure Characteristics of Feed Proteins and Protein Digestibility and Solubility

    Directory of Open Access Journals (Sweden)

    Mingmei Bai

    2016-08-01

    Full Text Available The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller’s dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as α-helices and β-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area of feed proteins were correlated with their the in vitro digestibility and solubility (p≤0.003; moreover, the relatively quantitative amounts of α-helices, random coils, and α-helix to β-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility (p≤0.004. On the other hand, the percentage of β-sheet structures was negatively correlated with protein in vitro digestibility (p<0.001 and solubility (p = 0.002. These results demonstrate that the molecular structure characteristics of feed proteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the α-helix-to-β-sheet ratio can be used to predict the nutritional value of feed proteins.

  7. THE SURFACE-MEDIATED UNFOLDING KINETICS OF GLOBULAR PROTEINS IS DEPENDENT ON MOLECULAR WEIGHT AND TEMPERATURE

    Energy Technology Data Exchange (ETDEWEB)

    Patananan, A.N.; Goheen, S.C.

    2008-01-01

    The adsorption and unfolding pathways of proteins on rigid surfaces are essential in numerous complex processes associated with biomedical engineering, nanotechnology, and chromatography. It is now well accepted that the kinetics of unfolding are characterized by chemical and physical interactions dependent on protein deformability and structure, as well as environmental pH, temperature, and surface chemistry. Although this fundamental process has broad implications in medicine and industry, little is known about the mechanism because of the atomic lengths and rapid time scales involved. Therefore, the unfolding kinetics of myoglobin, β-glucosidase, and ovalbumin were investigated by adsorbing the globular proteins to non-porous cationic polymer beads. The protein fractions were adsorbed at different residence times (0, 9, 10, 20, and 30 min) at near-physiological conditions using a gradient elution system similar to that in high-performance liquid chromatography. The elution profi les and retention times were obtained by ultraviolet/visible spectrophotometry. A decrease in recovery was observed with time for almost all proteins and was attributed to irreversible protein unfolding on the non-porous surfaces. These data, and those of previous studies, fi t a positively increasing linear trend between percent unfolding after a fi xed (9 min) residence time (71.8%, 31.1%, and 32.1% of myoglobin, β-glucosidase, and ovalbumin, respectively) and molecular weight. Of all the proteins examined so far, only myoglobin deviated from this trend with higher than predicted unfolding rates. Myoglobin also exhibited an increase in retention time over a wide temperature range (0°C and 55°C, 4.39 min and 5.74 min, respectively) whereas ovalbumin and β-glucosidase did not. Further studies using a larger set of proteins are required to better understand the physiological and physiochemical implications of protein unfolding kinetics. This study confi rms that surface

  8. Computational and theoretical studies of globular proteins

    Science.gov (United States)

    Pagan, Daniel L.

    Protein crystallization is often achieved in experiment through a trial and error approach. To date, there exists a dearth of theoretical understanding of the initial conditions necessary to promote crystallization. While a better understanding of crystallization will help to create good crystals suitable for structure analysis, it will also allow us to prevent the onset of certain diseases. The core of this thesis is to model and, ultimately, understand the phase behavior of protein particles in solution. Toward this goal, we calculate the fluid-fluid coexistence curve in the vicinity of the metastable critical point of the modified Lennard-Jones potential, where it has been shown that nucleation is increased by many orders of magnitude. We use finite-size scaling techniques and grand canonical Monte Carlo simulation methods. This has allowed us to pinpoint the critical point and subcritical region with high accuracy in spite of the critical fluctuations that hinder sampling using other Monte Carlo techniques. We also attempt to model the phase behavior of the gamma-crystallins, mutations of which have been linked to genetic cataracts. The complete phase behavior of the square well potential at the ranges of attraction lambda = 1.15 and lambda = 1.25 is calculated and compared with that of the gammaII-crystallin. The role of solvent is also important in the crystallization process and affects the phase behavior of proteins in solution. We study a model that accounts for the contribution of the solvent free-energy to the free-energy of globular proteins. This model allows us to model phase behavior that includes solvent.

  9. The direct piezoelectric effect in the globular protein lysozyme

    Science.gov (United States)

    Stapleton, A.; Noor, M. R.; Sweeney, J.; Casey, V.; Kholkin, A. L.; Silien, C.; Gandhi, A. A.; Soulimane, T.; Tofail, S. A. M.

    2017-10-01

    Here, we present experimental evidence of the direct piezoelectric effect in the globular protein, lysozyme. Piezoelectric materials are employed in many actuating and sensing applications because they can convert mechanical energy into electrical energy and vice versa. Although originally studied in inorganic materials, several biological materials including amino acids and bone, also exhibit piezoelectricity. The exact mechanisms supporting biological piezoelectricity are not known, nor is it known whether biological piezoelectricity conforms strictly to the criteria of classical piezoelectricity. The observation of piezoelectricity in protein crystals presented here links biological piezoelectricity with the classical theory of piezoelectricity. We quantify the direct piezoelectric effect in monoclinic and tetragonal aggregate films of lysozyme using conventional techniques based on the Berlincourt Method. The largest piezoelectric effect measured in a crystalline aggregate film of lysozyme was approximately 6.5 pC N-1. These findings raise fundamental questions as to the possible physiological significance of piezoelectricity in lysozyme and the potential for technical applications.

  10. Lysine Rich Proteins in the Salt-Soluble Protein Fraction of Barley

    DEFF Research Database (Denmark)

    Ingversen, J.; Køie, B.

    1973-01-01

    Fractionation of the protein complex from Emir barley showed that the salt-soluble fraction accounts for 44% of the total lysine content but only for 2.......Fractionation of the protein complex from Emir barley showed that the salt-soluble fraction accounts for 44% of the total lysine content but only for 2....

  11. SANS study of understanding mechanism of cold gelation of globular proteins

    International Nuclear Information System (INIS)

    Chinchalikar, A. J.; Kumar, Sugam; Aswal, V. K.; Wagh, A. G.; Kohlbrecher, J.

    2014-01-01

    Small-angle neutron scattering (SANS) has been used to probe the evolution of interaction and the resultant structures in the cold gelation of globular proteins. The cold gelation involves two steps consisting of irreversible protein deformation by heating followed by some means (e.g. increasing ionic strength) to bring them together at room temperature. We have examined the role of different salts in cold gelation of preheated aqueous Bovine Serum Albumin (BSA) protein solutions. The interactions have been modeled by two Yukawa potential combining short-range attraction and long-range repulsion. We show that in step 1 (preheated temperature effect) the deformation of protein increases the magnitude of attractive interaction but not sufficient to induce gel. The attractive interaction is further enhanced in step 2 (salt effect) to result in gel formation. The salt effect is found to be strongly depending on the valency of the counterions. The gel structure has been characterized by the mass fractals

  12. Dynamics of proteins and of their hydration layer studied by neutron scattering and additional biophysical methods

    International Nuclear Information System (INIS)

    Gallat, Francois-Xavier

    2011-01-01

    This thesis work focused on the dynamics of proteins, surrounded by their hydration layer, a water shell around the protein vital for its biological function. Each of these components is accompanied by a specific dynamics which union reforms the complex energy landscape of the system. The joint implementation of selective deuteration, incoherent neutron scattering and tera-hertz spectroscopy allowed to explore the dynamics of proteins and that of the hydration shell. The influence of the folding state of protein on its dynamics has been studied by elastic neutron scattering. Globular proteins were less dynamic than its intrinsically disordered analogues. Themselves appear to be stiffer than non-physiological unfolded proteins. The oligomerization state and the consequences on the dynamics were investigated. Aggregates of a globular protein proved to be more flexible than the soluble form. In contrast, aggregates of a disordered protein showed lower average dynamics compared to the soluble form. These observations demonstrate the wide range of dynamics among the proteome. Incoherent neutron scattering experiences on the hydration layer of globular and disordered proteins have yielded information on the nature of water motion around these proteins. The measurements revealed the presence of translational motions concomitant with the onset of the transition dynamics of hydration layers, at 220 K. Measurements have also shown a stronger coupling between a disordered protein and its hydration water, compared to a globular protein and its hydration shell. The nature of the hydration layer and its influence on its dynamics has been explored with the use of polymers that mimic the water behavior and that act as a source of flexibility for the protein. Eventually, the dynamics of methyl groups involved in the dynamical changes observed at 150 and 220 K, was investigated. (author) [fr

  13. Two novel heat-soluble protein families abundantly expressed in an anhydrobiotic tardigrade.

    Directory of Open Access Journals (Sweden)

    Ayami Yamaguchi

    Full Text Available Tardigrades are able to tolerate almost complete dehydration by reversibly switching to an ametabolic state. This ability is called anhydrobiosis. In the anhydrobiotic state, tardigrades can withstand various extreme environments including space, but their molecular basis remains largely unknown. Late embryogenesis abundant (LEA proteins are heat-soluble proteins and can prevent protein-aggregation in dehydrated conditions in other anhydrobiotic organisms, but their relevance to tardigrade anhydrobiosis is not clarified. In this study, we focused on the heat-soluble property characteristic of LEA proteins and conducted heat-soluble proteomics using an anhydrobiotic tardigrade. Our heat-soluble proteomics identified five abundant heat-soluble proteins. All of them showed no sequence similarity with LEA proteins and formed two novel protein families with distinct subcellular localizations. We named them Cytoplasmic Abundant Heat Soluble (CAHS and Secretory Abundant Heat Soluble (SAHS protein families, according to their localization. Both protein families were conserved among tardigrades, but not found in other phyla. Although CAHS protein was intrinsically unstructured and SAHS protein was rich in β-structure in the hydrated condition, proteins in both families changed their conformation to an α-helical structure in water-deficient conditions as LEA proteins do. Two conserved repeats of 19-mer motifs in CAHS proteins were capable to form amphiphilic stripes in α-helices, suggesting their roles as molecular shield in water-deficient condition, though charge distribution pattern in α-helices were different between CAHS and LEA proteins. Tardigrades might have evolved novel protein families with a heat-soluble property and this study revealed a novel repertoire of major heat-soluble proteins in these anhydrobiotic animals.

  14. Relationship between Molecular Structure Characteristics of Feed Proteins and Protein In vitro Digestibility and Solubility.

    Science.gov (United States)

    Bai, Mingmei; Qin, Guixin; Sun, Zewei; Long, Guohui

    2016-08-01

    The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as α-helices and β-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility (p≤0.003); moreover, the relatively quantitative amounts of α-helices, random coils, and α-helix to β-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility (p≤0.004). On the other hand, the percentage of β-sheet structures was negatively correlated with protein in vitro digestibility (pdigestibility at 28 h and solubility. Furthermore, the α-helix-to-β-sheet ratio can be used to predict the nutritional value of feed proteins.

  15. Facilitating protein solubility by use of peptide extensions

    Science.gov (United States)

    Freimuth, Paul I; Zhang, Yian-Biao; Howitt, Jason

    2013-09-17

    Expression vectors for expression of a protein or polypeptide of interest as a fusion product composed of the protein or polypeptide of interest fused at one terminus to a solubility enhancing peptide extension are provided. Sequences encoding the peptide extensions are provided. The invention further comprises antibodies which bind specifically to one or more of the solubility enhancing peptide extensions.

  16. EFFECT OF HEAT TREATMENT ON SOYBEAN PROTEIN SOLUBILITY

    Directory of Open Access Journals (Sweden)

    RODICA CĂPRIŢĂ

    2007-05-01

    Full Text Available The use of soybean products in animal feeds is limited due to the presence of antinutritional factors (ANF. Proper heat processing is required to destroy ANF naturally present in raw soybeans and to remove solvent remaining from the oil extraction process. Over and under toasting of soybean causes lower nutritional value. Excessive heat treatment causes Maillard reaction which affects the availability of lysine in particular and produces changes to the chemical structure of proteins resulting in a decrease of the nutritive value. The objective of this study was to evaluate the effect of heating time on the protein solubility. The investigation of the heating time on protein solubility in soybean meal (SBM revealed a negative correlation (r = -0.9596. Since the urease index is suitable only for detecting under processed SBM, the protein solubility is an important index for monitoring SBM quality.

  17. Self assembling nanocomposites for protein delivery: supramolecular interactions of soluble polymers with protein drugs.

    Science.gov (United States)

    Salmaso, Stefano; Caliceti, Paolo

    2013-01-02

    Translation of therapeutic proteins to pharmaceutical products is often encumbered by their inadequate physicochemical and biopharmaceutical properties, namely low stability and poor bioavailability. Over the last decades, several academic and industrial research programs have been focused on development of biocompatible polymers to produce appropriate formulations that provide for enhanced therapeutic performance. According to their physicochemical properties, polymers have been exploited to obtain a variety of formulations including biodegradable microparticles, 3-dimensional hydrogels, bioconjugates and soluble nanocomposites. Several soluble polymers bearing charges or hydrophobic moieties along the macromolecular backbone have been found to physically associate with proteins to form soluble nanocomplexes. Physical complexation is deemed a valuable alternative tool to the chemical bioconjugation. Soluble protein/polymer nanocomplexes formed by physical specific or unspecific interactions have been found in fact to possess peculiar physicochemical, and biopharmaceutical properties. Accordingly, soluble polymeric systems have been developed to increase the protein stability, enhance the bioavailability, promote the absorption across the biological barriers, and prolong the protein residence in the bloodstream. Furthermore, a few polymers have been found to favour the protein internalisation into cells or boost their immunogenic potential by acting as immunoadjuvant in vaccination protocols. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Membrane Proteins Are Dramatically Less Conserved than Water-Soluble Proteins across the Tree of Life.

    Science.gov (United States)

    Sojo, Victor; Dessimoz, Christophe; Pomiankowski, Andrew; Lane, Nick

    2016-11-01

    Membrane proteins are crucial in transport, signaling, bioenergetics, catalysis, and as drug targets. Here, we show that membrane proteins have dramatically fewer detectable orthologs than water-soluble proteins, less than half in most species analyzed. This sparse distribution could reflect rapid divergence or gene loss. We find that both mechanisms operate. First, membrane proteins evolve faster than water-soluble proteins, particularly in their exterior-facing portions. Second, we demonstrate that predicted ancestral membrane proteins are preferentially lost compared with water-soluble proteins in closely related species of archaea and bacteria. These patterns are consistent across the whole tree of life, and in each of the three domains of archaea, bacteria, and eukaryotes. Our findings point to a fundamental evolutionary principle: membrane proteins evolve faster due to stronger adaptive selection in changing environments, whereas cytosolic proteins are under more stringent purifying selection in the homeostatic interior of the cell. This effect should be strongest in prokaryotes, weaker in unicellular eukaryotes (with intracellular membranes), and weakest in multicellular eukaryotes (with extracellular homeostasis). We demonstrate that this is indeed the case. Similarly, we show that extracellular water-soluble proteins exhibit an even stronger pattern of low homology than membrane proteins. These striking differences in conservation of membrane proteins versus water-soluble proteins have important implications for evolution and medicine. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  19. Proteolytic activities of kiwifruit actinidin (Actinidia deliciosa cv. Hayward) on different fibrous and globular proteins: a comparative study of actinidin with papain.

    Science.gov (United States)

    Chalabi, Maryam; Khademi, Fatemeh; Yarani, Reza; Mostafaie, Ali

    2014-04-01

    Actinidin, a member of the papain-like family of cysteine proteases, is abundant in kiwifruit. To date, a few studies have been provided to investigate the proteolytic activity and substrate specificity of actinidin on native proteins. Herein, the proteolytic activity of actinidin was compared to papain on several different fibrous and globular proteins under neutral, acidic and basic conditions. The digested samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and densitometry to assess the proteolytic effect. Furthermore, the levels of free amino nitrogen (FAN) of the treated samples were determined using the ninhydrin colorimetric method. The findings showed that actinidin has no or limited proteolytic effect on globular proteins such as immunoglobulins including sheep IgG, rabbit IgG, chicken IgY and fish IgM, bovine serum albumin (BSA), lipid transfer protein (LTP), and whey proteins (α-lactalbumin and β-lactoglobulin) compared to papain. In contrast to globular proteins, actinidin could hydrolyze collagen and fibrinogen perfectly at neutral and mild basic pHs. Moreover, this enzyme could digest pure α-casein and major subunits of micellar casein especially in acidic pHs. Taken together, the data indicated that actinidin has narrow substrate specificity with the highest enzymatic activity for the collagen and fibrinogen substrates. The results describe the actinidin as a mild plant protease useful for many special applications such as cell isolation from different tissues and some food industries as a mixture formula with other relevant proteases.

  20. A Soluble, Folded Protein without Charged Amino Acid Residues

    DEFF Research Database (Denmark)

    Højgaard, Casper; Kofoed, Christian; Espersen, Roall

    2016-01-01

    Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable...... side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find...

  1. Conformational and functional analysis of the C-terminal globular head of the reovirus cell attachment protein.

    Science.gov (United States)

    Duncan, R; Horne, D; Strong, J E; Leone, G; Pon, R T; Yeung, M C; Lee, P W

    1991-06-01

    We have been investigating structure-function relationships in the reovirus cell attachment protein sigma 1 using various deletion mutants and protease analysis. In the present study, a series of deletion mutants were constructed which lacked 90, 44, 30, 12, or 4 amino acids from the C-terminus of the 455-amino acid-long reovirus type 3 (T3) sigma 1 protein. The full-length and truncated sigma 1 proteins were expressed in an in vitro transcription/translation system and assayed for L cell binding activity. It was found that the removal of as few as four amino acids from the C-terminus drastically affected the cell binding function of the sigma 1 protein. The C-terminal-truncated proteins were further characterized using trypsin, chymotrypsin, and monoclonal and polyclonal antibodies. Our results indicated that the C-terminal portions of the mutant proteins were misfolded, leading to a loss in cell binding function. The N-terminal fibrous tail of the proteins was unaffected by the deletions as was sigma 1 oligomerization, further illustrating the discrete structural and functional roles of the N- and C-terminal domains of sigma 1. In an attempt to identify smaller, functional peptides, full-length sigma 1 expressed in vitro was digested with trypsin and subsequently with chymotrypsin under various conditions. The results clearly demonstrated the highly stable nature of the C-terminal globular head of sigma 1, even when separated from the N-terminal fibrous tail. We concluded that: (1) the C-terminal globular head of sigma 1 exists as a compact, protease-resistant oligomeric structure; (2) an intact C-terminus is required for proper head folding and generation of the conformationally dependent cell binding domain.

  2. Protein solubility and folding enhancement by interaction with RNA.

    Directory of Open Access Journals (Sweden)

    Seong Il Choi

    Full Text Available While basic mechanisms of several major molecular chaperones are well understood, this machinery has been known to be involved in folding of only limited number of proteins inside the cells. Here, we report a chaperone type of protein folding facilitated by interaction with RNA. When an RNA-binding module is placed at the N-terminus of aggregation-prone target proteins, this module, upon binding with RNA, further promotes the solubility of passenger proteins, potentially leading to enhancement of proper protein folding. Studies on in vitro refolding in the presence of RNA, coexpression of RNA molecules in vivo and the mutants with impaired RNA binding ability suggests that RNA can exert chaperoning effect on their bound proteins. The results suggest that RNA binding could affect the overall kinetic network of protein folding pathway in favor of productive folding over off-pathway aggregation. In addition, the RNA binding-mediated solubility enhancement is extremely robust for increasing soluble yield of passenger proteins and could be usefully implemented for high-throughput protein expression for functional and structural genomic research initiatives. The RNA-mediated chaperone type presented here would give new insights into de novo folding in vivo.

  3. Optimization of translation profiles enhances protein expression and solubility.

    Directory of Open Access Journals (Sweden)

    Anne-Katrin Hess

    Full Text Available mRNA is translated with a non-uniform speed that actively coordinates co-translational folding of protein domains. Using structure-based homology we identified the structural domains in epoxide hydrolases (EHs and introduced slow-translating codons to delineate the translation of single domains. These changes in translation speed dramatically improved the solubility of two EHs of metagenomic origin in Escherichia coli. Conversely, the importance of transient attenuation for the folding, and consequently solubility, of EH was evidenced with a member of the EH family from Agrobacterium radiobacter, which partitions in the soluble fraction when expressed in E. coli. Synonymous substitutions of codons shaping the slow-transiting regions to fast-translating codons render this protein insoluble. Furthermore, we show that low protein yield can be enhanced by decreasing the free folding energy of the initial 5'-coding region, which can disrupt mRNA secondary structure and enhance ribosomal loading. This study provides direct experimental evidence that mRNA is not a mere messenger for translation of codons into amino acids but bears an additional layer of information for folding, solubility and expression level of the encoded protein. Furthermore, it provides a general frame on how to modulate and fine-tune gene expression of a target protein.

  4. Protein Solubility as Quality Index for Processed Soybean

    Directory of Open Access Journals (Sweden)

    Rodica Căpriţă

    2010-05-01

    Full Text Available Protein quality of soybean meal (SBM is linked to both the reduction of antinutritional factors (ANFs, and the optimization of protein digestibility. Both insufficient- and over-heating result in poor quality SBM. Inadequate heating fails to completely destroy the ANFs, which may have a detrimental impact on animal performance, while excessive heating reduces the availability of lysine via the Maillard reaction and possibly, to a lesser extent, of other amino acids. The objective of our study was to compare some biochemical laboratory procedures for assessing quality of SBM: urease index (UI, protein dispersibility index (PDI, KOH protein solubility (PS, and nitrogen solubility index (NSI. The experimental data reveal that UI is not useful to determine excessive heat treatment since additional heating has no effect on the urease index. KOH protein solubility remains high, during initial heat treatment. In marked contrast, the PDI and NSI decreased incrementally from 78% to 20% and from 97% to 60%, respectively, when heating 0 to 30 minutes. Combing the PDI test with the urease test could be useful to monitor soybean quality. SBM containing low UI (0.3 or below and high PDI (40 to 45% may indicate that the sample is definitely high quality because it has been adequately heat processed, but not overprocessed.

  5. Small-angle X-ray scattering studies of thermally-induced globular protein gels

    International Nuclear Information System (INIS)

    Clark, A.H.; Tuffnell, C.D.

    1980-01-01

    Small-angle X-ray scattering has been applied to gels formed by heating globular proteins, in aqueous solution, above their unfolding temperatures. A number of BSA gels, previously characterised by electron microscopy, have been studied, and by setting up theoretical models for the scattering process, the X-ray data have been shown to be consistent with the microscope conclusions regarding network structure. It is concluded that the networks form by a linearly-directed aggregation of unfolded, disc-like, protein molecules, three-dimensional geometry being achieved by occasional branching, and/or cross-linking. Long-range inhomogeneities in network structure, easily observed by electron microscopy, and correlated with variations in pH or ionic strength, have an effect on X-ray scattering, and hence the X-ray method is sensitive not only to different network strand thicknesses, but to different degrees of uniformity as well. (author)

  6. Self-assembly dynamics for the transition of a globular aggregate to a fibril network of lysozyme proteins via a coarse-grained Monte Carlo simulation

    Directory of Open Access Journals (Sweden)

    R. B. Pandey

    2015-09-01

    Full Text Available The self-organizing dynamics of lysozymes (an amyloid protein with 148 residues with different numbers of protein chains, Nc = 1,5,10, and 15 (concentration 0.004 – 0.063 is studied by a coarse-grained Monte Carlo simulation with knowledge-based residue-residue interactions. The dynamics of an isolated lysozyme (Nc = 1 is ultra-slow (quasi-static at low temperatures and becomes diffusive asymptotically on raising the temperature. In contrast, the presence of interacting proteins leads to concentration induced protein diffusion at low temperatures and concentration-tempering sub-diffusion at high temperatures. Variation of the radius of gyration of the protein with temperature shows a systematic transition from a globular structure (at low T to a random coil (high T conformation when the proteins are isolated. The crossover from globular to random coil becomes sharper upon increasing the protein concentration (i.e. with Nc = 5,10, with larger Rg at higher temperatures and concentration; Rg becomes smaller on adding more protein chains (e.g. Nc = 15 a non-monotonic response to protein concentration. Analysis of the structure factor (S(q provides an estimate of the effective dimension (D ≥ 3, globular conformation at low temperature, and D ∼ 1.7, random coil, at high temperatures of the isolated protein. With many interacting proteins, the morphology of the self-assembly varies with scale, i.e. at the low temperature (T = 0.015, D ∼ 2.9 on the scale comparable to the radius of gyration of the protein, and D ∼ 2.3 at the large scale over the entire sample. The global network of fibrils appears at high temperature (T = 0.021 with D ∼ 1.7 (i.e. a random coil morphology at large scale involving tenuous distribution of micro-globules (at small scales.

  7. Globular clusters - Fads and fallacies

    International Nuclear Information System (INIS)

    White, R.E.

    1991-01-01

    The types of globular clusters observed in the Milky Way Galaxy are described together with their known characteristics, with special attention given to correcting the erroneous statements made earlier about globular clusters. Among these are the following statements: the Galaxy is surrounded by many hundreds of globular clusters; all globular clusters are located toward the Galactic center, all globular clusters are metal poor and move about the Galaxy in highly elliptical paths; all globular clusters contain RR Lyrae-type variable stars, and the RR Lyrae stars found outside of globulars have come from cluster dissolution or ejection; all of the stars in a given cluster were born at the same time and have the same chemical composition; X-ray globulars are powered by central black holes; and the luminosity functions for globular clusters are well defined and well determined. Consideration is given to the fact that globular clusters in the Magellanic Clouds differ from those in the Milky Way by their age distribution and that the globulars of the SMC differ from those of the LMC

  8. AFM-based identification of the dynamic properties of globular proteins: simulation study

    International Nuclear Information System (INIS)

    Kim, Deok Ho; Park, Jung Yul; Kim, Moon K.; Hong, Keum Shik

    2008-01-01

    Nowadays a mathematical model-based computational approach is getting more attention as an effective tool for understanding the mechanical behaviors of biological systems. To find the mechanical properties of the proteins required to build such a model, this paper investigates a real-time identification method based on an AFM nanomanipulation system. First, an AFM-based bio-characterization system is introduced. Second, a second-order time-varying linear model representing the interaction between an AFM cantilever and globular proteins in a solvent is presented. Finally, we address a real-time estimation method in which the results of AFM experiments are designed to be inputs of the state estimator proposed here. Our attention is restricted to a theoretical feasibility analysis of the proposed methodology. We simply set the mechanical properties of the particular protein such as mass, stiffness, and damping coefficient in the system model prior to running the simulation. Simulation results show very good agreement with the preset properties. We anticipate that the realization of the AFM-based bio-characterization system will also provide an experimental validation of the proposed identification procedure in the future. This methodology can be used to determine a model of protein motion for the purpose of computer simulation and for a real-time modification of protein deformation

  9. PON-Sol: prediction of effects of amino acid substitutions on protein solubility.

    Science.gov (United States)

    Yang, Yang; Niroula, Abhishek; Shen, Bairong; Vihinen, Mauno

    2016-07-01

    Solubility is one of the fundamental protein properties. It is of great interest because of its relevance to protein expression. Reduced solubility and protein aggregation are also associated with many diseases. We collected from literature the largest experimentally verified solubility affecting amino acid substitution (AAS) dataset and used it to train a predictor called PON-Sol. The predictor can distinguish both solubility decreasing and increasing variants from those not affecting solubility. PON-Sol has normalized correct prediction ratio of 0.491 on cross-validation and 0.432 for independent test set. The performance of the method was compared both to solubility and aggregation predictors and found to be superior. PON-Sol can be used for the prediction of effects of disease-related substitutions, effects on heterologous recombinant protein expression and enhanced crystallizability. One application is to investigate effects of all possible AASs in a protein to aid protein engineering. PON-Sol is freely available at http://structure.bmc.lu.se/PON-Sol The training and test data are available at http://structure.bmc.lu.se/VariBench/ponsol.php mauno.vihinen@med.lu.se Supplementary data are available at Bioinformatics online. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  10. A Soluble, Folded Protein without Charged Amino Acid Residues

    DEFF Research Database (Denmark)

    Højgaard, Casper; Kofoed, Christian; Espersen, Roall

    2016-01-01

    side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find......Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable...... that the protein shows a surprising resilience toward extremes of pH, demonstrating stability and function (cellulose binding) in the pH range from 2 to 11. To ask whether the four charged residues present were required for these properties of this protein, we altered them to nontitratable ones. Remarkably...

  11. The nature of folded states of globular proteins.

    Science.gov (United States)

    Honeycutt, J D; Thirumalai, D

    1992-06-01

    We suggest, using dynamical simulations of a simple heteropolymer modelling the alpha-carbon sequence in a protein, that generically the folded states of globular proteins correspond to statistically well-defined metastable states. This hypothesis, called the metastability hypothesis, states that there are several free energy minima separated by barriers of various heights such that the folded conformations of a polypeptide chain in each of the minima have similar structural characteristics but have different energies from one another. The calculated structural characteristics, such as bond angle and dihedral angle distribution functions, are assumed to arise from only those configurations belonging to a given minimum. The validity of this hypothesis is illustrated by simulations of a continuum model of a heteropolymer whose low temperature state is a well-defined beta-barrel structure. The simulations were done using a molecular dynamics algorithm (referred to as the "noisy" molecular dynamics method) containing both friction and noise terms. It is shown that for this model there are several distinct metastable minima in which the structural features are similar. Several new methods of analyzing fluctuations in structures belonging to two distinct minima are introduced. The most notable one is a dynamic measure of compactness that can in principle provide the time required for maximal compactness to be achieved. The analysis shows that for a given metastable state in which the protein has a well-defined folded structure the transition to a state of higher compactness occurs very slowly, lending credence to the notion that the system encounters a late barrier in the process of folding to the most compact structure. The examination of the fluctuations in the structures near the unfolding----folding transition temperature indicates that the transition state for the unfolding to folding process occurs closer to the folded state.

  12. Efficient protein production method for NMR using soluble protein tags with cold shock expression vector

    International Nuclear Information System (INIS)

    Hayashi, Kokoro; Kojima, Chojiro

    2010-01-01

    The E. coli protein expression system is one of the most useful methods employed for NMR sample preparation. However, the production of some recombinant proteins in E. coli is often hampered by difficulties such as low expression level and low solubility. To address these problems, a modified cold-shock expression system containing a glutathione S-transferase (GST) tag, the pCold-GST system, was investigated. The pCold-GST system successfully expressed 9 out of 10 proteins that otherwise could not be expressed using a conventional E. coli expression system. Here, we applied the pCold-GST system to 84 proteins and 78 proteins were successfully expressed in the soluble fraction. Three other cold-shock expression systems containing a maltose binding protein tag (pCold-MBP), protein G B1 domain tag (pCold-GB1) or thioredoxin tag (pCold-Trx) were also developed to improve the yield. Additionally, we show that a C-terminal proline tag, which is invisible in 1 H- 15 N HSQC spectra, inhibits protein degradation and increases the final yield of unstable proteins. The purified proteins were amenable to NMR analyses. These data suggest that pCold expression systems combined with soluble protein tags can be utilized to improve the expression and purification of various proteins for NMR analysis.

  13. Limitations of polyethylene glycol-induced precipitation as predictive tool for protein solubility during formulation development.

    Science.gov (United States)

    Hofmann, Melanie; Winzer, Matthias; Weber, Christian; Gieseler, Henning

    2018-05-01

    Polyethylene glycol (PEG)-induced protein precipitation is often used to extrapolate apparent protein solubility at specific formulation compositions. The procedure was used for several fields of application such as protein crystal growth but also protein formulation development. Nevertheless, most studies focused on applicability in protein crystal growth. In contrast, this study focuses on applicability of PEG-induced precipitation during high-concentration protein formulation development. In this study, solubility of three different model proteins was investigated over a broad range of pH. Solubility values predicted by PEG-induced precipitation were compared to real solubility behaviour determined by either turbidity or content measurements. Predicted solubility by PEG-induced precipitation was confirmed for an Fc fusion protein and a monoclonal antibody. In contrast, PEG-induced precipitation failed to predict solubility of a single-domain antibody construct. Applicability of PEG-induced precipitation as indicator of protein solubility during formulation development was found to be not valid for one of three model molecules. Under certain conditions, PEG-induced protein precipitation is not valid for prediction of real protein solubility behaviour. The procedure should be used carefully as tool for formulation development, and the results obtained should be validated by additional investigations. © 2017 Royal Pharmaceutical Society.

  14. The Influence of Tissue Lyophilization and Gamma Irradiation on the Solubility of Proteins

    International Nuclear Information System (INIS)

    Komender, J.; Jendyk, J.; Leibschang, J.

    1967-01-01

    Most recent methods of tissue preservation are based on lyophilization and sterilization with gamma rays. Unfortunately the tissues preserved by this method lose their viability, this being connected with protein denaturation. The denaturing influence either of lyophilization or sterilization with gamma rays on different materials has been described. However, no observations on denaturation of proteins in prepared grafts are known. The work aimed at establishing the influence of individual stages of the procedure used in a tissue bank on the solubility of proteins. An experiment was performed using rat liver as a model tissue. Solubility of protein was determined in five groups of material, as follows: (1) fresh tissue, used as a control, (2) frozen tissue, (3) frozen and lyophilized tissue, (4) frozen, lyophilized and irradiated tissue, and (5) fresh irradiated tissue. Folin's method was used for determination of protein in water extracts of tissues. It was found that: (1) the whole procedure considerably diminished the protein solubility, (2) freezing diminishes the protein solubility by 35% on average, (3) lyophilization causes no further protein denaturation, (4) protein solubility is reduced most (by about 65%) by sterilization with gamma rays. (author)

  15. Protein fiber linear dichroism for structure determination and kinetics in a low-volume, low-wavelength couette flow cell

    OpenAIRE

    Dafforn, Tim; Rajendra, Jacindra; Halsall, David J.; Serpell, Louise C.; Rodger, Alison

    2004-01-01

    High-resolution structure determination of soluble globular proteins relies heavily on x-ray crystallography techniques. Such an approach is often ineffective for investigations into the structure of fibrous proteins as these proteins generally do not crystallize. Thus investigations into fibrous protein structure have relied on less direct methods such as x-ray fiber diffraction and circular dichroism. Ultraviolet linear dichroism has the potential to provide additional information on the st...

  16. High-level expression of soluble recombinant proteins in Escherichia coli using an HE-maltotriose-binding protein fusion tag.

    Science.gov (United States)

    Han, Yingqian; Guo, Wanying; Su, Bingqian; Guo, Yujie; Wang, Jiang; Chu, Beibei; Yang, Guoyu

    2018-02-01

    Recombinant proteins are commonly expressed in prokaryotic expression systems for large-scale production. The use of genetically engineered affinity and solubility enhancing fusion proteins has increased greatly in recent years, and there now exists a considerable repertoire of these that can be used to enhance the expression, stability, solubility, folding, and purification of their fusion partner. Here, a modified histidine tag (HE) used as an affinity tag was employed together with a truncated maltotriose-binding protein (MBP; consisting of residues 59-433) from Pyrococcus furiosus as a solubility enhancing tag accompanying a tobacco etch virus protease-recognition site for protein expression and purification in Escherichia coli. Various proteins tagged at the N-terminus with HE-MBP(Pyr) were expressed in E. coli BL21(DE3) cells to determine expression and solubility relative to those tagged with His6-MBP or His6-MBP(Pyr). Furthermore, four HE-MBP(Pyr)-fused proteins were purified by immobilized metal affinity chromatography to assess the affinity of HE with immobilized Ni 2+ . Our results showed that HE-MBP(Pyr) represents an attractive fusion protein allowing high levels of soluble expression and purification of recombinant protein in E. coli. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Efficient protein production method for NMR using soluble protein tags with cold shock expression vector

    Energy Technology Data Exchange (ETDEWEB)

    Hayashi, Kokoro [Fujifilm Corporation, Analysis Technology Center (Japan); Kojima, Chojiro, E-mail: kojima@protein.osaka-u.ac.j [Nara Institute of Science and Technology (NAIST), Graduate School of Biological Sciences (Japan)

    2010-11-15

    The E. coli protein expression system is one of the most useful methods employed for NMR sample preparation. However, the production of some recombinant proteins in E. coli is often hampered by difficulties such as low expression level and low solubility. To address these problems, a modified cold-shock expression system containing a glutathione S-transferase (GST) tag, the pCold-GST system, was investigated. The pCold-GST system successfully expressed 9 out of 10 proteins that otherwise could not be expressed using a conventional E. coli expression system. Here, we applied the pCold-GST system to 84 proteins and 78 proteins were successfully expressed in the soluble fraction. Three other cold-shock expression systems containing a maltose binding protein tag (pCold-MBP), protein G B1 domain tag (pCold-GB1) or thioredoxin tag (pCold-Trx) were also developed to improve the yield. Additionally, we show that a C-terminal proline tag, which is invisible in {sup 1}H-{sup 15}N HSQC spectra, inhibits protein degradation and increases the final yield of unstable proteins. The purified proteins were amenable to NMR analyses. These data suggest that pCold expression systems combined with soluble protein tags can be utilized to improve the expression and purification of various proteins for NMR analysis.

  18. A single cysteine post-translational oxidation suffices to compromise globular proteins kinetic stability and promote amyloid formation

    Directory of Open Access Journals (Sweden)

    Patrizia Marinelli

    2018-04-01

    Full Text Available Oxidatively modified forms of proteins accumulate during aging. Oxidized protein conformers might act as intermediates in the formation of amyloids in age-related disorders. However, it is not known whether this amyloidogenic conversion requires an extensive protein oxidative damage or it can be promoted just by a discrete, localized post-translational modification of certain residues. Here, we demonstrate that the irreversible oxidation of a single free Cys suffices to severely perturb the folding energy landscape of a stable globular protein, compromise its kinetic stability, and lead to the formation of amyloids under physiological conditions. Experiments and simulations converge to indicate that this specific oxidation-promoted protein aggregation requires only local unfolding. Indeed, a large scale analysis indicates that many cellular proteins are at risk of undergoing this kind of deleterious transition; explaining how oxidative stress can impact cell proteostasis and subsequently lead to the onset of pathological states. Keywords: Protein oxidation, Protein misfolding, Protein aggregation, Oxidative stress, Post-translational modification

  19. Toward the description of electrostatic interactions between globular proteins: potential of mean force in the primitive model.

    Science.gov (United States)

    Dahirel, Vincent; Jardat, Marie; Dufrêche, Jean-François; Turq, Pierre

    2007-09-07

    Monte Carlo simulations are used to calculate the exact potential of mean force between charged globular proteins in aqueous solution. The aim of the present paper is to study the influence of the ions of the added salt on the effective interaction between these nanoparticles. The charges of the model proteins, either identical or opposite, are either central or distributed on a discrete pattern. Contrarily to Poisson-Boltzmann predictions, attractive, and repulsive direct forces between proteins are not screened similarly. Moreover, it has been shown that the relative orientations of the charge patterns strongly influence salt-mediated interactions. More precisely, for short distances between the proteins, ions enhance the difference of the effective forces between (i) like-charged and oppositely charged proteins, (ii) attractive and repulsive relative orientations of the proteins, which may affect the selectivity of protein/protein recognition. Finally, such results observed with the simplest models are applied to a more elaborate one to demonstrate their generality.

  20. Globular clusters, old and young

    International Nuclear Information System (INIS)

    Samus', N.N.

    1984-01-01

    The problem of similarity of and difference in the globular and scattered star clusters is considered. Star clusters in astronomy are related either to globular or to scattered ones according to the structure of Hertzsprung-Russell diagram constructed for star clusters, but not according to the appearance. The qlobular clusters in the Galaxy are composed of giants and subgiants, which testifies to the old age of the globular clusters. The Globular clusters in the Magellanic clouds are classified into ''red'' ones - similar to the globular clusters of the Galaxy, and ''blue'' ones - similar to them in appearance but differing extremely by the star composition and so by the age. The old star clusters are suggested to be called globular ones, while another name (''populous'', for example) is suggested to be used for other clusters similar to globular ones only in appearance

  1. Larger red-shift in optical emissions obtained from the thin films of globular proteins (BSA, lysozyme) – polyelectrolyte (PAA) complexes

    Energy Technology Data Exchange (ETDEWEB)

    Talukdar, Hrishikesh [Physical Sciences Division, Institute of Advanced Study in Science and Technology, Vigyan Path, Paschim Boragaon, Garchuk, Guwahati 781035, Assam (India); Kundu, Sarathi, E-mail: sarathi.kundu@gmail.com [Physical Sciences Division, Institute of Advanced Study in Science and Technology, Vigyan Path, Paschim Boragaon, Garchuk, Guwahati 781035, Assam (India); Basu, Saibal [Solid State Physics Division, Bhabha Atomic Research Centre, Mumbai 400 085 (India)

    2016-09-30

    Graphical abstract: Thin films of protein-polyelectrolyte complexes show larger red-shift in optical emission. - Highlights: • Globular proteins (lysozyme and BSA) and polyelectrolyte (sodium polyacrylic acid) are used to form protein-polyelectrolyte complexes (PPC). • Larger red-shift in optical emission is obtained from the thin films of PPC. • Red-shift is not obtained from the solution of PPC and pure protein thin films. • Larger red-shift from PPC films is due to the energy dissipation as non-radiative form through interactions with nearby atoms. • Red-shift in optical emission is independent on the thickness of the PPC film. - Abstract: Globular proteins (lysozyme and BSA) and polyelectrolyte (sodium polyacrylic acid) are used to form protein-polyelectrolyte complexes (PPC). Out-of-plane structures of ≈30–60 nm thick PPC films and their surface morphologies have been studied by using X-ray reflectivity and atomic force microscopy, whereas optical behaviors of PPC and protein conformations have been studied by using UV–vis, photoluminescence and FTIR spectroscopy respectively. Our study reveals that thin films of PPC show a larger red-shift of 23 and 16 nm in the optical emissions in comparison to that of pure protein whereas bulk PPC show a small blue-shift of ≈3 nm. A small amount of peak-shift is found to occur due to the heat treatment or concentration variation of the polyelectrolyte/protein in bulk solution but cannot produce such film thickness independent larger red-shift. Position of the emission peak remains nearly unchanged with the film thickness. Mechanism for such larger red-shift has been proposed.

  2. Globular Clusters - Guides to Galaxies

    CERN Document Server

    Richtler, Tom; Joint ESO-FONDAP Workshop on Globular Clusters

    2009-01-01

    The principal question of whether and how globular clusters can contribute to a better understanding of galaxy formation and evolution is perhaps the main driving force behind the overall endeavour of studying globular cluster systems. Naturally, this splits up into many individual problems. The objective of the Joint ESO-FONDAP Workshop on Globular Clusters - Guides to Galaxies was to bring together researchers, both observational and theoretical, to present and discuss the most recent results. Topics covered in these proceedings are: internal dynamics of globular clusters and interaction with host galaxies (tidal tails, evolution of cluster masses), accretion of globular clusters, detailed descriptions of nearby cluster systems, ultracompact dwarfs, formations of massive clusters in mergers and elsewhere, the ACS Virgo survey, galaxy formation and globular clusters, dynamics and kinematics of globular cluster systems and dark matter-related problems. With its wide coverage of the topic, this book constitute...

  3. Soluble Non-ammonia Nitrogen in Ruminal and Omasal Digesta of Korean Native Steers Supplemented with Soluble Proteins

    Directory of Open Access Journals (Sweden)

    C. W. Choi

    2012-09-01

    Full Text Available An experiment was conducted to study the effect of soluble protein supplements on concentration of soluble non-ammonia nitrogen (SNAN in the liquid phase of ruminal (RD and omasal digesta (OD of Korean native steers, and to investigate diurnal pattern in SNAN concentration in RD and OD. Three ruminally cannulated Korean native steers in a 3×3 Latin square design consumed a basal diet of rice straw and corn-based concentrate (control, and that supplemented (kg/d DM basis with intact casein (0.24; IC or acid hydrolyzed casein (0.46; AHC. Ruminal digesta was sampled using a vacuum pump, whereas OD was collected using an omasal sampling system at 2.0 h intervals after a morning feeding. The SNAN fractions (free amino acid (AA, peptide and soluble protein in RD and OD were assessed using the ninhydrin assay. Concentrations of free AA and total SNAN in RD were significantly (p<0.05 lower than those in OD. Although free AA concentration was relatively high, mean peptide was quantitatively the most important fraction of total SNAN in both RD and OD, indicating that degradation of peptide to AA rather than hydrolysis of soluble protein to peptide or deamination may be the most limiting step in rumen proteolysis of Korean native steers. Diurnal variation in peptide concentration in OD for the soluble protein supplemented diets during the feeding cycle peaked 2 h post-feeding and decreased thereafter whereas that for the control was relatively constant during the entire feeding cycle. Diurnal variation in peptide concentration was rather similar between RD and OD.

  4. Salt-bridge networks within globular and disordered proteins: characterizing trends for designable interactions.

    Science.gov (United States)

    Basu, Sankar; Mukharjee, Debasish

    2017-07-01

    There has been considerable debate about the contribution of salt bridges to the stabilization of protein folds, in spite of their participation in crucial protein functions. Salt bridges appear to contribute to the activity-stability trade-off within proteins by bringing high-entropy charged amino acids into close contacts during the course of their functions. The current study analyzes the modes of association of salt bridges (in terms of networks) within globular proteins and at protein-protein interfaces. While the most common and trivial type of salt bridge is the isolated salt bridge, bifurcated salt bridge appears to be a distinct salt-bridge motif having a special topology and geometry. Bifurcated salt bridges are found ubiquitously in proteins and interprotein complexes. Interesting and attractive examples presenting different modes of interaction are highlighted. Bifurcated salt bridges appear to function as molecular clips that are used to stitch together large surface contours at interacting protein interfaces. The present work also emphasizes the key role of salt-bridge-mediated interactions in the partial folding of proteins containing long stretches of disordered regions. Salt-bridge-mediated interactions seem to be pivotal to the promotion of "disorder-to-order" transitions in small disordered protein fragments and their stabilization upon binding. The results obtained in this work should help to guide efforts to elucidate the modus operandi of these partially disordered proteins, and to conceptualize how these proteins manage to maintain the required amount of disorder even in their bound forms. This work could also potentially facilitate explorations of geometrically specific designable salt bridges through the characterization of composite salt-bridge networks. Graphical abstract ᅟ.

  5. Description of atomic burials in compact globular proteins by Fermi-Dirac probability distributions.

    Science.gov (United States)

    Gomes, Antonio L C; de Rezende, Júlia R; Pereira de Araújo, Antônio F; Shakhnovich, Eugene I

    2007-02-01

    We perform a statistical analysis of atomic distributions as a function of the distance R from the molecular geometrical center in a nonredundant set of compact globular proteins. The number of atoms increases quadratically for small R, indicating a constant average density inside the core, reaches a maximum at a size-dependent distance R(max), and falls rapidly for larger R. The empirical curves turn out to be consistent with the volume increase of spherical concentric solid shells and a Fermi-Dirac distribution in which the distance R plays the role of an effective atomic energy epsilon(R) = R. The effective chemical potential mu governing the distribution increases with the number of residues, reflecting the size of the protein globule, while the temperature parameter beta decreases. Interestingly, betamu is not as strongly dependent on protein size and appears to be tuned to maintain approximately half of the atoms in the high density interior and the other half in the exterior region of rapidly decreasing density. A normalized size-independent distribution was obtained for the atomic probability as a function of the reduced distance, r = R/R(g), where R(g) is the radius of gyration. The global normalized Fermi distribution, F(r), can be reasonably decomposed in Fermi-like subdistributions for different atomic types tau, F(tau)(r), with Sigma(tau)F(tau)(r) = F(r), which depend on two additional parameters mu(tau) and h(tau). The chemical potential mu(tau) affects a scaling prefactor and depends on the overall frequency of the corresponding atomic type, while the maximum position of the subdistribution is determined by h(tau), which appears in a type-dependent atomic effective energy, epsilon(tau)(r) = h(tau)r, and is strongly correlated to available hydrophobicity scales. Better adjustments are obtained when the effective energy is not assumed to be necessarily linear, or epsilon(tau)*(r) = h(tau)*r(alpha,), in which case a correlation with hydrophobicity

  6. Novel mitochondria-targeted heat-soluble proteins identified in the anhydrobiotic Tardigrade improve osmotic tolerance of human cells.

    Directory of Open Access Journals (Sweden)

    Sae Tanaka

    Full Text Available Tardigrades are able to tolerate almost complete dehydration through transition to a metabolically inactive state, called "anhydrobiosis". Late Embryogenesis Abundant (LEA proteins are heat-soluble proteins involved in the desiccation tolerance of many anhydrobiotic organisms. Tardigrades, Ramazzottius varieornatus, however, express predominantly tardigrade-unique heat-soluble proteins: CAHS (Cytoplasmic Abundant Heat Soluble and SAHS (Secretory Abundant Heat Soluble proteins, which are secreted or localized in most intracellular compartments, except the mitochondria. Although mitochondrial integrity is crucial to ensure cellular survival, protective molecules for mitochondria have remained elusive. Here, we identified two novel mitochondrial heat-soluble proteins, RvLEAM and MAHS (Mitochondrial Abundant Heat Soluble, as potent mitochondrial protectants from Ramazzottius varieornatus. RvLEAM is a group3 LEA protein and immunohistochemistry confirmed its mitochondrial localization in tardigrade cells. MAHS-green fluorescent protein fusion protein localized in human mitochondria and was heat-soluble in vitro, though no sequence similarity with other known proteins was found, and one region was conserved among tardigrades. Furthermore, we demonstrated that RvLEAM protein as well as MAHS protein improved the hyperosmotic tolerance of human cells. The findings of the present study revealed that tardigrade mitochondria contain at least two types of heat-soluble proteins that might have protective roles in water-deficient environments.

  7. Profiling and relationship of water-soluble sugar and protein compositions in soybean seeds.

    Science.gov (United States)

    Yu, Xiaomin; Yuan, Fengjie; Fu, Xujun; Zhu, Danhua

    2016-04-01

    Sugar and protein are important quality traits in soybean seeds for making soy-based food products. However, the investigations on both compositions and their relationship have rarely been reported. In this study, a total of 35 soybean germplasms collected from Zhejiang province of China, were evaluated for both water-soluble sugar and protein. The total water-soluble sugar (TWSS) content of the germplasms studied ranged from 84.70 to 140.91 mg/g and the water-soluble protein (WSP) content varied from 26.5% to 36.0%. The WSP content showed positive correlations with the TWSS and sucrose contents but negative correlations with the fructose and glucose contents. The clustering showed the 35 germplasms could be divided into four groups with specific contents of sugar and protein. The combination of water-soluble sugar and protein profiles provides useful information for future breeding and genetic research. This investigation will facilitate future work for seed quality improvement. Copyright © 2015. Published by Elsevier Ltd.

  8. BtcA, A class IA type III chaperone, interacts with the BteA N-terminal domain through a globular/non-globular mechanism.

    Directory of Open Access Journals (Sweden)

    Chen Guttman

    Full Text Available Bordetella pertussis, the etiological agent of "whooping cough" disease, utilizes the type III secretion system (T3SS to deliver a 69 kDa cytotoxic effector protein, BteA, directly into the host cells. As with other T3SS effectors, prior to its secretion BteA binds BtcA, a 13.9 kDa protein predicted to act as a T3SS class IA chaperone. While this interaction had been characterized for such effector-chaperone pairs in other pathogens, it has yet to be fully investigated in Bordetella. Here we provide the first biochemical proof that BtcA is indeed a class IA chaperone, responsible for the binding of BteA's N-terminal domain. We bring forth extensive evidence that BtcA binds its substrate effector through a dual-interface binding mechanism comprising of non-globular and bi-globular interactions at a moderate micromolar level binding affinity. We demonstrate that the non-globular interactions involve the first 31 N-terminal residues of BteA287 and their removal leads to destabilization of the effector-chaperone complex and lower binding affinities to BtcA. These findings represent an important first step towards a molecular understanding of BteA secretion and cell entry.

  9. Optics of globular photonic crystals

    International Nuclear Information System (INIS)

    Gorelik, V S

    2007-01-01

    The results of experimental and theoretical studies of the optical properties of globular photonic crystals - new physical objects having a crystal structure with the lattice period exceeding considerably the atomic size, are presented. As globular photonic crystals, artificial opal matrices consisting of close-packed silica globules of diameter ∼200 nm were used. The reflection spectra of these objects characterising the parameters of photonic bands existing in these crystals in the visible spectral region are presented. The idealised models of the energy band structure of photonic crystals investigated in the review give analytic dispersion dependences for the group velocity and the effective photon mass in a globular photonic crystal. The characteristics of secondary emission excited in globular photonic crystals by monochromatic and broadband radiation are presented. The results of investigations of single-photon-excited delayed scattering of light observed in globular photonic crystals exposed to cw UV radiation and radiation from a repetitively pulsed copper vapour laser are presented. The possibilities of using globular photonic crystals as active media for lasing in different spectral regions are considered. It is proposed to use globular photonic crystals as sensitive sensors in optoelectronic devices for molecular analysis of organic and inorganic materials by the modern methods of laser spectroscopy. The results of experimental studies of spontaneous and stimulated globular scattering of light are discussed. The conditions for observing resonance and two-photon-excited delayed scattering of light are found. The possibility of accumulation and localisation of the laser radiation energy inside a globular photonic crystal is reported. (review)

  10. Soluble protein isolated from low cost fish and fish wastes

    OpenAIRE

    Lekshmy Nair, A.; Gopakumar, K.

    1982-01-01

    The method of preparation, composition, amino acid content, protein efficiency ratio and areas of possible application of water soluble protein isolates from low cost fish and fish wastes are discussed in detail in this communication.

  11. Effects of gamma irradiation on physicochemical properties of heat-induced gel prepared with chicken salt-soluble proteins

    International Nuclear Information System (INIS)

    Choi, Yun-Sang; Kim, Hyun-Wook; Hwang, Ko-Eun; Song, Dong-Heon; Jeong, Tae-Jun; Seo, Kwang-Wook; Kim, Young-Boong; Kim, Cheon-Jei

    2015-01-01

    The technological effects of gamma irradiation (0, 3, 7, and 10 kGy) on chicken salt-soluble meat proteins in a model system were investigated. There were no significant differences in protein, fat, and ash content, and sarcoplasmic protein solubility among all samples. The samples with increasing gamma irradiation levels had higher pH, lightness, yellowness, and apparent viscosity, whereas moisture content, water holding capacity, redness, myofibrillar protein solubility, total protein solubility, hardness, springiness, cohesiveness, gumminess, and chewiness were the highest in the unirradiated control. The result from meat products using gamma irradiation was intended to provide a basic resource processing technology. - Highlights: • The effect of gamma irradiation on salt-soluble meat proteins was investigated. • Gelling properties of salt-soluble protein affected by gamma irradiation. • Gamma irradiation of meat products provides a basic resource processing technology

  12. Changes in protein solubility, fermentative capacity, viscoelasticity ...

    African Journals Online (AJOL)

    Frozen dough should be stored for fewer than 21 days; time in which the loaf volume of bread made from frozen dough was approximately 40.84% smaller than that of fresh bread dough formulation. Keywords: French type bread, frozen dough, protein solubility, baking quality, viscoelasticity. African Journal of Biotechnology ...

  13. REVIEW: Optics of globular photonic crystals

    Science.gov (United States)

    Gorelik, V. S.

    2007-05-01

    The results of experimental and theoretical studies of the optical properties of globular photonic crystals - new physical objects having a crystal structure with the lattice period exceeding considerably the atomic size, are presented. As globular photonic crystals, artificial opal matrices consisting of close-packed silica globules of diameter ~200 nm were used. The reflection spectra of these objects characterising the parameters of photonic bands existing in these crystals in the visible spectral region are presented. The idealised models of the energy band structure of photonic crystals investigated in the review give analytic dispersion dependences for the group velocity and the effective photon mass in a globular photonic crystal. The characteristics of secondary emission excited in globular photonic crystals by monochromatic and broadband radiation are presented. The results of investigations of single-photon-excited delayed scattering of light observed in globular photonic crystals exposed to cw UV radiation and radiation from a repetitively pulsed copper vapour laser are presented. The possibilities of using globular photonic crystals as active media for lasing in different spectral regions are considered. It is proposed to use globular photonic crystals as sensitive sensors in optoelectronic devices for molecular analysis of organic and inorganic materials by the modern methods of laser spectroscopy. The results of experimental studies of spontaneous and stimulated globular scattering of light are discussed. The conditions for observing resonance and two-photon-excited delayed scattering of light are found. The possibility of accumulation and localisation of the laser radiation energy inside a globular photonic crystal is reported.

  14. Soluble Milk Protein Supplementation with Moderate Physical Activity Improves Locomotion Function in Aging Rats.

    Directory of Open Access Journals (Sweden)

    Aude Lafoux

    Full Text Available Aging is associated with a loss of muscle mass and functional capacity. Present study was designed to compare the impact of specific dairy proteins on muscular function with or without a low-intensity physical activity program on a treadmill in an aged rat model. We investigated the effects of nutritional supplementation, five days a week over a 2-month period with a slow digestible protein, casein or fast digestible proteins, whey or soluble milk protein, on strength and locomotor parameters in sedentary or active aged Wistar RjHan rats (17-19 months of age. An extensive gait analysis was performed before and after protein supplementation. After two months of protein administration and activity program, muscle force was evaluated using a grip test, spontaneous activity using an open-field and muscular mass by specific muscle sampling. When aged rats were supplemented with proteins without exercise, only minor effects of different diets on muscle mass and locomotion were observed: higher muscle mass in the casein group and improvement of stride frequencies with soluble milk protein. By contrast, supplementation with soluble milk protein just after physical activity was more effective at improving overall skeletal muscle function in old rats compared to casein. For active old rats supplemented with soluble milk protein, an increase in locomotor activity in the open field and an enhancement of static and dynamic gait parameters compared to active groups supplemented with casein or whey were observed without any differences in muscle mass and forelimb strength. These results suggest that consumption of soluble milk protein as a bolus immediately after a low intensity physical activity may be a suitable nutritional intervention to prevent decline in locomotion in aged rats and strengthen the interest to analyze the longitudinal aspect of locomotion in aged rodents.

  15. Soluble Milk Protein Supplementation with Moderate Physical Activity Improves Locomotion Function in Aging Rats.

    Science.gov (United States)

    Lafoux, Aude; Baudry, Charlotte; Bonhomme, Cécile; Le Ruyet, Pascale; Huchet, Corinne

    2016-01-01

    Aging is associated with a loss of muscle mass and functional capacity. Present study was designed to compare the impact of specific dairy proteins on muscular function with or without a low-intensity physical activity program on a treadmill in an aged rat model. We investigated the effects of nutritional supplementation, five days a week over a 2-month period with a slow digestible protein, casein or fast digestible proteins, whey or soluble milk protein, on strength and locomotor parameters in sedentary or active aged Wistar RjHan rats (17-19 months of age). An extensive gait analysis was performed before and after protein supplementation. After two months of protein administration and activity program, muscle force was evaluated using a grip test, spontaneous activity using an open-field and muscular mass by specific muscle sampling. When aged rats were supplemented with proteins without exercise, only minor effects of different diets on muscle mass and locomotion were observed: higher muscle mass in the casein group and improvement of stride frequencies with soluble milk protein. By contrast, supplementation with soluble milk protein just after physical activity was more effective at improving overall skeletal muscle function in old rats compared to casein. For active old rats supplemented with soluble milk protein, an increase in locomotor activity in the open field and an enhancement of static and dynamic gait parameters compared to active groups supplemented with casein or whey were observed without any differences in muscle mass and forelimb strength. These results suggest that consumption of soluble milk protein as a bolus immediately after a low intensity physical activity may be a suitable nutritional intervention to prevent decline in locomotion in aged rats and strengthen the interest to analyze the longitudinal aspect of locomotion in aged rodents.

  16. Effect of protein solution components in the adsorption of Herbaspirillum seropedicae GlnB protein on mica.

    Science.gov (United States)

    Ferreira, Cecília F G; Benelli, Elaine M; Klein, Jorge J; Schreiner, Wido; Camargo, Paulo C

    2009-10-15

    The adsorption of proteins and its buffer solution on mica surfaces was investigated by atomic force microscopy (AFM). Different salt concentration of the Herbaspirillum seropedicae GlnB protein (GlnB-Hs) solution deposited on mica was investigated. This protein is a globular, soluble homotrimer (36kDa), member of PII-like proteins family involved in signal transducing in prokaryote. Supramolecular structures were formed when this protein was deposited onto bare mica surface. The topographic AFM images of the GlnB-Hs films showed that at high salt concentration the supramolecular structures are spherical-like, instead of the typical doughnut-like shape for low salt concentration. AFM images of NaCl and Tris from the buffer solution showed structures with the same pattern as those observed for high salt protein solution, misleading the image interpretation. XPS experiments showed that GlnB protein film covers the mica surface without chemical reaction.

  17. Relativistic Binaries in Globular Clusters

    Directory of Open Access Journals (Sweden)

    Matthew J. Benacquista

    2013-03-01

    Full Text Available Galactic globular clusters are old, dense star systems typically containing 10^4 – 10^6 stars. As an old population of stars, globular clusters contain many collapsed and degenerate objects. As a dense population of stars, globular clusters are the scene of many interesting close dynamical interactions between stars. These dynamical interactions can alter the evolution of individual stars and can produce tight binary systems containing one or two compact objects. In this review, we discuss theoretical models of globular cluster evolution and binary evolution, techniques for simulating this evolution that leads to relativistic binaries, and current and possible future observational evidence for this population. Our discussion of globular cluster evolution will focus on the processes that boost the production of tight binary systems and the subsequent interaction of these binaries that can alter the properties of both bodies and can lead to exotic objects. Direct N-body integrations and Fokker–Planck simulations of the evolution of globular clusters that incorporate tidal interactions and lead to predictions of relativistic binary populations are also discussed. We discuss the current observational evidence for cataclysmic variables, millisecond pulsars, and low-mass X-ray binaries as well as possible future detection of relativistic binaries with gravitational radiation.

  18. Photometric studies of globular clusters in the Andromeda Nebula. Luminosity function for old globular clusters

    International Nuclear Information System (INIS)

    Sharov, A.S.; Lyutyj, V.M.

    1989-01-01

    The luminosity function for old globular clusters in M 31 is presented. The objects were selected according to their structural and photometric properties. At the usually accepted normal (Gaussian) distribution, the luminosity function is characterized by the following parameters: the mean magnitude, corrected for the extinction inside M 31, V-bar 0 =16 m ,38±0 m .08, and the absolute magnitude M-bar v =-8 m .29 assuming )m-M) v =23 m .67, standard deviation σ M v =1 m .16±0 m .08 and total object number N=300±17. Old globular clusters in M 31 are in the average about one magnitude more luminous then those in our Galaxy (M v ≅ -7 m .3). Intrinsic luminosity dispersions of globular clusters are nearly the same in both galaxies. Available data on globular clusters in the Local Group galaxies against the universality of globular luminosity function with identical parameters M v and σ M v

  19. Identifying the adaptive mechanism in globular proteins: Fluctuations in densely packed regions manipulate flexible parts

    Science.gov (United States)

    Yilmaz, Lutfu Safak; Atilgan, Ali Rana

    2000-09-01

    A low-resolution structural model based on the packing geometry of α-carbons is utilized to establish a connection between the flexible and rigid parts of a folded protein. The former commonly recognizes a complementing molecule for making a complex, while the latter manipulates the necessary conformational change for binding. We attempt analytically to distinguish this control architecture that intrinsically exists in globular proteins. First with two-dimensional simple models, then for a native protein, bovine pancreatic trypsin inhibitor, we explicitly demonstrate that inserting fluctuations in tertiary contacts supported by the stable core, one can regulate the displacement of residues on loop regions. The positional fluctuations of the flexible regions are annihilated by the rest of the protein in conformity with the Le Chatelier-Braun principle. The results indicate that the distortion of the principal nonbonded contacts between highly packed residues is accompanied by that of the slavery fluctuations that are widely distributed over the native structure. These positional arrangements do not appear in a reciprocal relation between a perturbation and the associated response; the effect of a movement of residue i on residue j is not equal to that of the same movement of residue j on residue i.

  20. Screening of genetic parameters for soluble protein expression in Escherichia coli

    DEFF Research Database (Denmark)

    Vernet, Erik; Kotzsch, Alexander; Voldborg, Bjørn

    2011-01-01

    Soluble expression of proteins in a relevant form for functional and structural investigations still often remains a challenge. Although many biochemical factors are known to affect solubility, a thorough investigation of yield-limiting factors is normally not feasible in high-throughput efforts....... Here we present a screening strategy for expression of biomedically relevant proteins in Escherichia coli using a panel of six different genetic variations. These include engineered strains for rare codon supplementation, increased disulfide bond formation in the cytoplasm and novel vectors...... for secretion to the periplasm or culture medium. Combining these variants with expression construct truncations design, we report on parallel cloning and expression of more than 300 constructs representing 24 selected proteins; including full-length variants of human growth factors, interleukins and growth...

  1. Globular clusters and galaxy halos

    International Nuclear Information System (INIS)

    Van Den Bergh, S.

    1984-01-01

    Using semipartial correlation coefficients and bootstrap techniques, a study is made of the important features of globular clusters with respect to the total number of galaxy clusters and dependence of specific galaxy cluster on parent galaxy type, cluster radii, luminosity functions and cluster ellipticity. It is shown that the ellipticity of LMC clusters correlates significantly with cluster luminosity functions, but not with cluster age. The cluter luminosity value above which globulars are noticeably flattened may differ by a factor of about 100 from galaxy to galaxy. Both in the Galaxy and in M31 globulars with small core radii have a Gaussian distribution over luminosity, whereas clusters with large core radii do not. In the cluster systems surrounding the Galaxy, M31 and NGC 5128 the mean radii of globular clusters was found to increase with the distance from the nucleus. Central galaxies in rich clusters have much higher values for specific globular cluster frequency than do other cluster ellipticals, suggesting that such central galaxies must already have been different from normal ellipticals at the time they were formed

  2. Physics of protein folding

    Science.gov (United States)

    Finkelstein, A. V.; Galzitskaya, O. V.

    2004-04-01

    Protein physics is grounded on three fundamental experimental facts: protein, this long heteropolymer, has a well defined compact three-dimensional structure; this structure can spontaneously arise from the unfolded protein chain in appropriate environment; and this structure is separated from the unfolded state of the chain by the “all-or-none” phase transition, which ensures robustness of protein structure and therefore of its action. The aim of this review is to consider modern understanding of physical principles of self-organization of protein structures and to overview such important features of this process, as finding out the unique protein structure among zillions alternatives, nucleation of the folding process and metastable folding intermediates. Towards this end we will consider the main experimental facts and simple, mostly phenomenological theoretical models. We will concentrate on relatively small (single-domain) water-soluble globular proteins (whose structure and especially folding are much better studied and understood than those of large or membrane and fibrous proteins) and consider kinetic and structural aspects of transition of initially unfolded protein chains into their final solid (“native”) 3D structures.

  3. Immunochemical analyses of soluble lens proteins in some marine fishes

    Digital Repository Service at National Institute of Oceanography (India)

    Menezes, M.R.

    Soluble eye lens proteins of 10 fishes, belonging to the families Clupeidae, Hemirhamphidae, Lactaridae, Scombridae, Stromatidae, Psettodidae, Bothidae and Soleidae were studied by immunoelectrophoresis using the lens antiserum of Sardinella...

  4. Production and characterization of cowpea protein hydrolysate with optimum nitrogen solubility by enzymatic hydrolysis using pepsin.

    Science.gov (United States)

    Mune Mune, Martin Alain; Minka, Samuel René

    2017-06-01

    Cowpea is a source of low-cost and good nutritional quality protein for utilization in food formulations in replacement of animal proteins. Therefore it is necessary that cowpea protein exhibits good functionality, particularly protein solubility which affects the other functional properties. The objective of this study was to produce cowpea protein hydrolysate exhibiting optimum solubility by the adequate combination of hydrolysis parameters, namely time, solid/liquid ratio (SLR) and enzyme/substrate ratio (ESR), and to determine its functional properties and molecular characteristics. A Box-Behnken experimental design was used for the experiments, and a second-order polynomial to model the effects of hydrolysis time, SLR and ESR on the degree of hydrolysis and nitrogen solubility index. The optimum hydrolysis conditions of time 208.61 min, SLR 1/15 (w/w) and ESR 2.25% (w/w) yielded a nitrogen solubility of 75.71%. Protein breakdown and the peptide profile following enzymatic hydrolysis were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and size exclusion chromatography. Cowpea protein hydrolysate showed higher oil absorption capacity, emulsifying activity and foaming ability compared with the concentrate. The solubility of cowpea protein hydrolysate was adequately optimized by response surface methodology, and the hydrolysate showed adequate functionality for use in food. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  5. Biochemical characterization of soluble proteins in pecan [Carya illinoinensis (Wangenh.) K. Koch].

    Science.gov (United States)

    Venkatachalam, Mahesh; Roux, Kenneth H; Sathe, Shridhar K

    2008-09-10

    Pecans (cv. Desirable) contained approximately 10% protein on a dry weight basis. The minimum nitrogen solubility (5.9-7.5%) at 0.25-0.75 M trichloroacetic acid represented the nonprotein nitrogen. Among the solvents assessed for protein solubilization, 0.1 M NaOH was the most effective, while borate saline buffer (pH 8.45) was judged to be optimal for protein solubilization. The protein solubility was minimal in the pH range of 3-7 and significantly increased on either side of this pH range. Increasing the NaCl concentration from 0 to 4 M significantly improved ( approximately 8-fold increase) protein solubilization. Following Osborne protein fractionation, the alkali-soluble glutelin fraction (60.1%) accounted for a major portion of pecan proteins followed by globulin (31.5%), prolamin (3.4%), and albumin (1.5%), respectively. The majority of pecan polypeptides were in the molecular mass range of 12-66 kDa and in the pI range of 4.0-8.3. The pecan globulin fraction was characterized by the presence of several glycoprotein polypeptides. Lysine was the first limiting essential amino acid in the defatted flour, globulin, prolamin, and alkaline glutelin fractions. Leucine and tryptophan were the first limiting essential amino acids in albumin and acid glutelin fractions, respectively. Rabbit polyclonal antibodies detected a range of pecan polypeptides in the 12-60 kDa range, of which the globulin fraction contained the most reactive polypeptides.

  6. Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression.

    Science.gov (United States)

    Chung, Jeong Min; Lee, Sangmin; Jung, Hyun Suk

    2017-05-01

    Bacterial expression is commonly used to produce recombinant and truncated mutant eukaryotic proteins. However, heterologous protein expression may render synthesized proteins insoluble. The conventional method used to express a poorly soluble protein, which involves denaturation and refolding, is time-consuming and inefficient. There are several non-denaturing approaches that can increase the solubility of recombinant proteins that include using different bacterial cell strains, altering the time of induction, lowering the incubation temperature, and employing different detergents for purification. In this study, we compared several non-denaturing protocols to express and purify two insoluble 34 kDa actin-bundling protein mutants. The solubility of the mutant proteins was not affected by any of the approaches except for treatment with the detergent sarkosyl. These results indicate that sarkosyl can effectively improve the solubility of insoluble proteins during bacterial expression. Copyright © 2016. Published by Elsevier Inc.

  7. Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay.

    Science.gov (United States)

    Asai, Akira; Takakuma, Kazuyuki

    2017-01-01

    When a large hydrophobic full-length protein is expressed in bacteria, it is often challenging to obtain recombinant proteins in the soluble fraction. One way to overcome this challenge is expression of deletion mutants that have improved solubility while maintaining biological activity. In this chapter, we describe a protocol for expression of truncated forms of STAT5b and STAT3 proteins that are soluble and retain SH2-mediated activity for phospho-Tyr peptide recognition.

  8. Common structural features of cholesterol binding sites in crystallized soluble proteins.

    Science.gov (United States)

    Bukiya, Anna N; Dopico, Alejandro M

    2017-06-01

    Cholesterol-protein interactions are essential for the architectural organization of cell membranes and for lipid metabolism. While cholesterol-sensing motifs in transmembrane proteins have been identified, little is known about cholesterol recognition by soluble proteins. We reviewed the structural characteristics of binding sites for cholesterol and cholesterol sulfate from crystallographic structures available in the Protein Data Bank. This analysis unveiled key features of cholesterol-binding sites that are present in either all or the majority of sites: i ) the cholesterol molecule is generally positioned between protein domains that have an organized secondary structure; ii ) the cholesterol hydroxyl/sulfo group is often partnered by Asn, Gln, and/or Tyr, while the hydrophobic part of cholesterol interacts with Leu, Ile, Val, and/or Phe; iii ) cholesterol hydrogen-bonding partners are often found on α-helices, while amino acids that interact with cholesterol's hydrophobic core have a slight preference for β-strands and secondary structure-lacking protein areas; iv ) the steroid's C21 and C26 constitute the "hot spots" most often seen for steroid-protein hydrophobic interactions; v ) common "cold spots" are C8-C10, C13, and C17, at which contacts with the proteins were not detected. Several common features we identified for soluble protein-steroid interaction appear evolutionarily conserved. Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.

  9. Production of a soluble recombinant prion protein fused to blue fluorescent protein without refolding or detergents in Escherichia coli cells.

    Science.gov (United States)

    Arii, Yasuhiro; Yamaguchi, Hidenori; Fukuoka, Shin-Ichi

    2007-10-01

    The physiological function of prion proteins (PrP) remains unclear. To investigate the physiological relevance of PrP, we constructed a fusion protein of PrP with enhanced blue fluorescent protein (PrP-EBFP) to quantify the interaction of PrP with other molecules. Production of soluble PrP-EBFP was achieved by lowering the expression temperature in Escherichia coli (E. coli) cells to 15 degrees C. Soluble PrP-EBFP was purified on cation exchange and heparin-affinity columns to yield high purity protein. This is the first report of the preparation of soluble recombinant PrP without refolding following solubilization using denaturants or disruption using detergents. To confirm the integrity of PrP-EBFP, anisotropy was estimated under physiological conditions in the presence of heparin, which interacts with PrP. The dissociation constant was determined to be 0.88+/-0.07 microM. PrP-EBFP should be useful in the quantification of PrP interactions with other molecules.

  10. Front-face fluorescence spectroscopy study of globular proteins in emulsions: displacement of BSA by a nonionic surfactant.

    Science.gov (United States)

    Rampon, V; Genot, C; Riaublanc, A; Anton, M; Axelos, M A V; McClements, D J

    2003-04-23

    The displacement of a globular protein (bovine serum albumin, BSA) from the surface of oil droplets in concentrated oil-in-water emulsions by a nonionic surfactant (polyoxyethylene sorbitan monolauarate, Tween 20) was studied using front-face fluorescence spectroscopy (FFFS). This method relies on measurement of the change in intensity (I(MAX)) and wavelength (lambda(MAX)) of the maximum in the tryptophan emission spectrum. A series of oil-in-water emulsions (21 wt % n-hexadecane, 0.22 wt % BSA, pH 7.0) containing different molar ratios of Tween 20 to BSA (R = 0-131) were prepared. As the surfactant concentration was increased, the protein was progressively displaced from the droplet surfaces. At R > or = 66, the protein was completely displaced from the droplet surfaces. There was an increase in both I(MAX) and lambda(MAX) with increasing Tween 20 concentration up to R = 66, which correlated with the increase in the ratio of nonadsorbed to adsorbed protein. In contrast, there was a decrease in I(MAX) and lambda(MAX) with Tween 20 concentration in protein solutions and for R > or = 66 in the emulsions, which was attributed to binding of the surfactant to the protein. This study shows that FFFS is a powerful technique for nondestructively providing information about the interfacial composition of droplets in concentrated protein-stabilized emulsions in situ. Nevertheless, in general the suitability of the technique may also depend on protein type and the nature of the physicochemical matrix surrounding the proteins.

  11. Synthesis of acid-soluble spore proteins by Bacillus subtilis.

    OpenAIRE

    Leventhal, J M; Chambliss, G H

    1982-01-01

    The major acid-soluble spore proteins (ASSPs) of Bacillus subtilis were detected by immunoprecipitation of radioactively labeled in vitro- and in vivo-synthesized proteins. ASSP synthesis in vivo began 2 h after the initiation of sporulation (t2) and reached its maximum rate at t7. This corresponded to the time of synthesis of mRNA that stimulated the maximum rate of ASSP synthesis in vitro. Under the set of conditions used in these experiments, protease synthesis began near t0, alkaline phos...

  12. Pregalactic formation of globular clusters in cold dark matter

    International Nuclear Information System (INIS)

    Faber, S.M.; Blumenthal, G.R.; Rosenblatt, E.I.

    1988-01-01

    The pregalactic hypothesis for the formation of globular clusters is reconsidered in the light of Zinn's (1985) discovery of a two-component globular population in the Milky Way. For a cold dark matter spectrum, high-sigma fluctuations of 10 to the 5th - 10 to the 6th solar masses are assumed to be the progenitors of the spheroidal population of globular clusters. The mass fraction of globular clusters in galaxies then requires that perturbations above roughly 2.8 sigma survive as globulars, and their observed radii require baryonic collapse factors of order 10. Such an absolute density threshold for globular cluster formation achieves adequate fits to observed cluster radii and densities, the mass fraction of globulars versus Hubble type, the radial density profile of globulars within galaxies, and the globular luminosity function. However, a fixed density threshold criterion for cluster survival lacks convincing physical justification and does not by itself explain the homogeneous metallicities within clusters or the large metallicity variations from cluster to cluster and from galaxy to galaxy. 33 references

  13. Soluble Protein Analysis using a Compact Bench-top Flow Cytometer

    Science.gov (United States)

    Pappas, Dimitri; Kao, Shib-Hsin; Cyr, Johnathan

    2004-01-01

    Future space exploration missions will require analytical technology capable of providing both autonomous medical care to the crew and investigative capabilities to researchers. While several promising candidate technologies exist for further development, flow cytometry is an attractive technology as it offers both crew health (blood cell count, leukocyte differential, etc.) and a wide array of biochemistry and immunology assays. research settings, the application of this technique to soluble protein analysis is also possible. Proteomic beads using fluorescent dyes for optical encoding were used to monitor six cytokines simultaneously in cell medium of cell cultures in stationary and rotating cell culture systems. The results of this work demonstrate that a compact flow cytometer, such as a system proposed for space flight, can detect a variety of soluble proteins for crew health and biotechnology experiments during long-term missions.

  14. A review of machine learning methods to predict the solubility of overexpressed recombinant proteins in Escherichia coli.

    Science.gov (United States)

    Habibi, Narjeskhatoon; Mohd Hashim, Siti Z; Norouzi, Alireza; Samian, Mohammed Razip

    2014-05-08

    Over the last 20 years in biotechnology, the production of recombinant proteins has been a crucial bioprocess in both biopharmaceutical and research arena in terms of human health, scientific impact and economic volume. Although logical strategies of genetic engineering have been established, protein overexpression is still an art. In particular, heterologous expression is often hindered by low level of production and frequent fail due to opaque reasons. The problem is accentuated because there is no generic solution available to enhance heterologous overexpression. For a given protein, the extent of its solubility can indicate the quality of its function. Over 30% of synthesized proteins are not soluble. In certain experimental circumstances, including temperature, expression host, etc., protein solubility is a feature eventually defined by its sequence. Until now, numerous methods based on machine learning are proposed to predict the solubility of protein merely from its amino acid sequence. In spite of the 20 years of research on the matter, no comprehensive review is available on the published methods. This paper presents an extensive review of the existing models to predict protein solubility in Escherichia coli recombinant protein overexpression system. The models are investigated and compared regarding the datasets used, features, feature selection methods, machine learning techniques and accuracy of prediction. A discussion on the models is provided at the end. This study aims to investigate extensively the machine learning based methods to predict recombinant protein solubility, so as to offer a general as well as a detailed understanding for researches in the field. Some of the models present acceptable prediction performances and convenient user interfaces. These models can be considered as valuable tools to predict recombinant protein overexpression results before performing real laboratory experiments, thus saving labour, time and cost.

  15. Semen quality and concentration of soluble proteins in the seminal plasma of Alpine bucks Semen quality and concentration of soluble proteins in the seminal plasma of Alpine bucks

    Directory of Open Access Journals (Sweden)

    Simone Eliza Facione Guimarães

    2010-06-01

    Full Text Available It was aimed to study the in vitro seminal quality analyzed by complementary tests and to compare them with physical, morphological and biochemical aspects of male goat semen of the Alpine breed. This experiment took place at the Federal University of Viçosa, situated at 20º45’ S latitude and 42º51’ W longitude, Southwest of Brazil. It was done during the summer months of January and February, and three adult male goats of the Alpine breed were used in intensive conditions. The semen was collected by artificial vagina method. In all semen samples (45 ejaculates, after the physical and morphological analysis, the hiposmotic test was done. In 24 ejaculates, it were done thermo-resistance test, and in 21 ejaculates it were determined the concentration of total soluble proteins in seminal plasma. The male goats presented difference in the semen physical and morphological aspects, in the hiposmotic test and thermo-resistance test, but they did not presented difference in total soluble proteins concentration in seminal plasma. Results of the slow thermo-resistance test and hiposmotic test were positively correlated (r = 0.60. It was concluded, according to our results, that the concentration of total soluble proteins in seminal plasma can not be used as a parameter to predict the seminal quality of Alpine bucks.It was aimed to study the in vitro seminal quality analyzed by complementary tests and to compare them with physical, morphological and biochemical aspects of male goat semen of the Alpine breed. This experiment took place at the Federal University of Viçosa, situated at 20º45’ S latitude and 42º51’ W longitude, Southwest of Brazil. It was done during the summer months of January and February, and three adult male goats of the Alpine breed were used in intensive conditions. The semen was collected by artificial vagina method. In all semen samples (45 ejaculates, after the physical and morphological analysis, the hiposmotic test

  16. Salt-soluble proteins from wheat-derived foodstuffs show lower allergenic potency than those from raw flour.

    Science.gov (United States)

    de Gregorio, Marta; Armentia, Alicia; Díaz-Perales, Araceli; Palacín, Arantxa; Dueñas-Laita, Antonio; Martín, Blanca; Salcedo, Gabriel; Sánchez-Monge, Rosa

    2009-04-22

    Salt-soluble proteins from wheat flour have been described as main allergens associated with both baker's asthma and food allergy. However, most studies have used raw flour as starting material, thus not considering potential changes in allergenic properties induced by the heat treatment and other industrial processing to produce wheat-derived foodstuffs. Salt extracts from different commercial wheat-derived products were obtained and their allergenic properties investigated by IgE-immunodetection, ELISA assays, and skin prick test. The IgE-binding capacity of salt-soluble proteins from commercial breads and cooked pastas was reduced around 50% compared with that of raw flour, the reduction being less dramatic in noncooked pastas and biscuits. Several wheat-derived foodstuffs showed major IgE-binding components of 20 and 35 kDa, identified as avenin-like and globulin proteins, respectively. These proteins, as well as most flour and bread salt-soluble proteins, were hydrolyzed when subjected to simulated gastrointestinal digestion. However, the digested products still exhibited a residual IgE-binding capacity. Therefore, processing of wheat flour to obtain derived foodstuffs decreases the IgE binding-capacity of the major salt-soluble wheat proteins. Moreover, simulated gastric fluid digestion further inactivates some heat-resistant IgE-binding proteins.

  17. Globular Clusters for Faint Galaxies

    Science.gov (United States)

    Kohler, Susanna

    2017-07-01

    The origin of ultra-diffuse galaxies (UDGs) has posed a long-standing mystery for astronomers. New observations of several of these faint giants with the Hubble Space Telescope are now lending support to one theory.Faint-Galaxy MysteryHubble images of Dragonfly 44 (top) and DFX1 (bottom). The right panels show the data with greater contrast and extended objects masked. [van Dokkum et al. 2017]UDGs large, extremely faint spheroidal objects were first discovered in the Virgo galaxy cluster roughly three decades ago. Modern telescope capabilities have resulted in many more discoveries of similar faint galaxies in recent years, suggesting that they are a much more common phenomenon than we originally thought.Despite the many observations, UDGs still pose a number of unanswered questions. Chief among them: what are UDGs? Why are these objects the size of normal galaxies, yet so dim? There are two primary models that explain UDGs:UDGs were originally small galaxies, hence their low luminosity. Tidal interactions then puffed them up to the large size we observe today.UDGs are effectively failed galaxies. They formed the same way as normal galaxies of their large size, but something truncated their star formation early, preventing them from gaining the brightness that we would expect for galaxies of their size.Now a team of scientists led by Pieter van Dokkum (Yale University) has made some intriguing observations with Hubble that lend weight to one of these models.Globulars observed in 16 Coma-cluster UDGs by Hubble. The top right panel shows the galaxy identifications. The top left panel shows the derived number of globular clusters in each galaxy. [van Dokkum et al. 2017]Globulars GaloreVan Dokkum and collaborators imaged two UDGs with Hubble: Dragonfly 44 and DFX1, both located in the Coma galaxy cluster. These faint galaxies are both smooth and elongated, with no obvious irregular features, spiral arms, star-forming regions, or other indications of tidal interactions

  18. THE EFFECT OF THE PROTEIN SOLUBILITY OF FISH MEAL AND ...

    African Journals Online (AJOL)

    utilization of fish meal proteins was determined by a relationship between their solubility or apparent digestibility and the ... Data obtained from these trials should indicate what re- ..... 4 in mind it is thus obvious that the big variation which oc-.

  19. Effect of pH adjustment, homogenization and diafiltration on physicochemical, reconstitution, functional and rheological properties of medium protein milk protein concentrates (MPC70).

    Science.gov (United States)

    Meena, Ganga Sahay; Singh, Ashish Kumar; Gupta, Vijay Kumar; Borad, Sanket; Arora, Sumit; Tomar, Sudhir Kumar

    2018-04-01

    Poor solubility is the major limiting factor in commercial applications of milk protein concentrates (MPC) powders. Retentate treatments such as pH adjustment using disodium phosphate (Na 2 HPO 4 ), also responsible for calcium chelation with homogenization and; its diafiltration with 150 mM NaCl solution were hypothesized to improve the functional properties of treated MPC70 powders. These treatments significantly improved the solubility, heat stability, water binding, dispersibility, bulk density, flowability, buffer index, foaming and emulsifying capacity of treated powders over control. Rheological behaviour of reconstituted MPC solutions was best explained by Herschel Bulkley model. Compared to rough, large globular structures with dents in control; majorly intact, separate, smaller particles of smooth surface, without any aggregation were observed in SEM micrograph of treated powders. Applied treatments are easy, cost-effective and capable to improve functional properties of treated powders that could replace control MPC70 powder in various food applications where protein functionality is of prime importance.

  20. Chromosomal rearrangements and protein globularity changes in Mycobacterium tuberculosis isolates from cerebrospinal fluid

    Directory of Open Access Journals (Sweden)

    Seow Hoon Saw

    2016-09-01

    Full Text Available Background Meningitis is a major cause of mortality in tuberculosis (TB. It is not clear what factors promote central nervous system invasion and pathology but it has been reported that certain strains of Mycobacterium tuberculosis (Mtb might have genetic traits associated with neurotropism. Methods In this study, we generated whole genome sequences of eight clinical strains of Mtb that were isolated from the cerebrospinal fluid (CSF of patients presenting with tuberculous meningitis (TBM in Malaysia, and compared them to the genomes of H37Rv and other respiratory Mtb genomes either downloaded from public databases or extracted from local sputum isolates. We aimed to find genomic features that might be distinctly different between CSF-derived and respiratory Mtb. Results Genome-wide comparisons revealed rearrangements (translocations, inversions, insertions and deletions and non-synonymous SNPs in our CSF-derived strains that were not observed in the respiratory Mtb genomes used for comparison. These rearranged segments were rich in genes for PE (proline-glutamate/PPE (proline-proline-glutamate, transcriptional and membrane proteins. Similarly, most of the ns SNPs common in CSF strains were noted in genes encoding PE/PPE proteins. Protein globularity differences were observed among mycobacteria from CSF and respiratory sources and in proteins previously reported to be associated with TB meningitis. Transcription factors and other transcription regulators featured prominently in these proteins. Homologs of proteins associated with Streptococcus pneumoniae meningitis and Neisseria meningitidis virulence were identified in neuropathogenic as well as respiratory mycobacterial spp. examined in this study. Discussion The occurrence of in silico genetic differences in CSF-derived but not respiratory Mtb suggests their possible involvement in the pathogenesis of TBM. However, overall findings in this comparative analysis support the postulation that TB

  1. Activation of human natural killer cells by the soluble form of cellular prion protein

    International Nuclear Information System (INIS)

    Seong, Yeon-Jae; Sung, Pil Soo; Jang, Young-Soon; Choi, Young Joon; Park, Bum-Chan; Park, Su-Hyung; Park, Young Woo; Shin, Eui-Cheol

    2015-01-01

    Cellular prion protein (PrP C ) is widely expressed in various cell types, including cells of the immune system. However, the specific roles of PrP C in the immune system have not been clearly elucidated. In the present study, we investigated the effects of a soluble form of recombinant PrP C protein on human natural killer (NK) cells. Recombinant soluble PrP C protein was generated by fusion of human PrP C with the Fc portion of human IgG 1 (PrP C -Fc). PrP C -Fc binds to the surface of human NK cells, particularly to CD56 dim NK cells. PrP C -Fc induced the production of cytokines and chemokines and the degranulation of granzyme B from NK cells. In addition, PrP C -Fc facilitated the IL-15-induced proliferation of NK cells. PrP C -Fc induced phosphorylation of ERK-1/2 and JNK in NK cells, and inhibitors of the ERK or the JNK pathways abrogated PrP C -Fc-induced cytokine production in NK cells. In conclusion, the soluble form of recombinant PrP C -Fc protein activates human NK cells via the ERK and JNK signaling pathways. - Highlights: • Recombinant soluble PrP C (PrP C -Fc) was generated by fusion of human PrP C with IgG1 Fc portion. • PrP C -Fc protein induces the production of cytokines and degranulation from human NK cells. • PrP C -Fc protein enhances the IL-15-induced proliferation of human NK cells. • PrP C -Fc protein activates human NK cells via the ERK and JNK signaling pathways

  2. Gamma-ray Emission from Globular Clusters

    Directory of Open Access Journals (Sweden)

    Pak-Hin T. Tam

    2016-03-01

    Full Text Available Over the last few years, the data obtained using the Large Area Telescope (LAT aboard the Fermi Gamma-ray Space Telescope has provided new insights on high-energy processes in globular clusters, particularly those involving compact objects such as MilliSecond Pulsars (MSPs. Gamma-ray emission in the 100 MeV to 10 GeV range has been detected from more than a dozen globular clusters in our galaxy, including 47 Tucanae and Terzan 5. Based on a sample of known gammaray globular clusters, the empirical relations between gamma-ray luminosity and properties of globular clusters such as their stellar encounter rate, metallicity, and possible optical and infrared photon energy densities, have been derived. The measured gamma-ray spectra are generally described by a power law with a cut-off at a few gigaelectronvolts. Together with the detection of pulsed γ-rays from two MSPs in two different globular clusters, such spectral signature lends support to the hypothesis that γ-rays from globular clusters represent collective curvature emission from magnetospheres of MSPs in the clusters. Alternative models, involving Inverse-Compton (IC emission of relativistic electrons that are accelerated close to MSPs or pulsar wind nebula shocks, have also been suggested. Observations at >100 GeV by using Fermi/LAT and atmospheric Cherenkov telescopes such as H.E.S.S.-II, MAGIC-II, VERITAS, and CTA will help to settle some questions unanswered by current data.

  3. The recombinant globular head domain of the measles virus hemagglutinin protein as a subunit vaccine against measles.

    Science.gov (United States)

    Lobanova, Liubov M; Eng, Nelson F; Satkunarajah, Malathy; Mutwiri, George K; Rini, James M; Zakhartchouk, Alexander N

    2012-04-26

    Despite the availability of live attenuated measles virus (MV) vaccines, a large number of measles-associated deaths occur among infants in developing countries. The development of a measles subunit vaccine may circumvent the limitations associated with the current live attenuated vaccines and eventually contribute to global measles eradication. Therefore, the goal of this study was to test the feasibility of producing the recombinant globular head domain of the MV hemagglutinin (H) protein by stably transfected human cells and to examine the ability of this recombinant protein to elicit MV-specific immune responses. The recombinant protein was purified from the culture supernatant of stably transfected HEK293T cells secreting a tagged version of the protein. Two subcutaneous immunizations with the purified recombinant protein alone resulted in the production of MV-specific serum IgG and neutralizing antibodies in mice. Formulation of the protein with adjuvants (polyphosphazene or alum) further enhanced the humoral immune response and in addition resulted in the induction of cell-mediated immunity as measured by the production of MV H-specific interferon gamma (IFN-γ) and interleukin 5 (IL-5) by in vitro re-stimulated splenocytes. Furthermore, the inclusion of polyphosphazene into the vaccine formulation induced a mixed Th1/Th2-type immune response. In addition, the purified recombinant protein retained its immunogenicity even after storage at 37°C for 2 weeks. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Predicting the activity coefficients of free-solvent for concentrated globular protein solutions using independently determined physical parameters.

    Directory of Open Access Journals (Sweden)

    Devin W McBride

    Full Text Available The activity coefficient is largely considered an empirical parameter that was traditionally introduced to correct the non-ideality observed in thermodynamic systems such as osmotic pressure. Here, the activity coefficient of free-solvent is related to physically realistic parameters and a mathematical expression is developed to directly predict the activity coefficients of free-solvent, for aqueous protein solutions up to near-saturation concentrations. The model is based on the free-solvent model, which has previously been shown to provide excellent prediction of the osmotic pressure of concentrated and crowded globular proteins in aqueous solutions up to near-saturation concentrations. Thus, this model uses only the independently determined, physically realizable quantities: mole fraction, solvent accessible surface area, and ion binding, in its prediction. Predictions are presented for the activity coefficients of free-solvent for near-saturated protein solutions containing either bovine serum albumin or hemoglobin. As a verification step, the predictability of the model for the activity coefficient of sucrose solutions was evaluated. The predicted activity coefficients of free-solvent are compared to the calculated activity coefficients of free-solvent based on osmotic pressure data. It is observed that the predicted activity coefficients are increasingly dependent on the solute-solvent parameters as the protein concentration increases to near-saturation concentrations.

  5. A family of E. coli expression vectors for laboratory scale and high throughput soluble protein production

    Directory of Open Access Journals (Sweden)

    Bottomley Stephen P

    2006-03-01

    Full Text Available Abstract Background In the past few years, both automated and manual high-throughput protein expression and purification has become an accessible means to rapidly screen and produce soluble proteins for structural and functional studies. However, many of the commercial vectors encoding different solubility tags require different cloning and purification steps for each vector, considerably slowing down expression screening. We have developed a set of E. coli expression vectors with different solubility tags that allow for parallel cloning from a single PCR product and can be purified using the same protocol. Results The set of E. coli expression vectors, encode for either a hexa-histidine tag or the three most commonly used solubility tags (GST, MBP, NusA and all with an N-terminal hexa-histidine sequence. The result is two-fold: the His-tag facilitates purification by immobilised metal affinity chromatography, whilst the fusion domains act primarily as solubility aids during expression, in addition to providing an optional purification step. We have also incorporated a TEV recognition sequence following the solubility tag domain, which allows for highly specific cleavage (using TEV protease of the fusion protein to yield native protein. These vectors are also designed for ligation-independent cloning and they possess a high-level expressing T7 promoter, which is suitable for auto-induction. To validate our vector system, we have cloned four different genes and also one gene into all four vectors and used small-scale expression and purification techniques. We demonstrate that the vectors are capable of high levels of expression and that efficient screening of new proteins can be readily achieved at the laboratory level. Conclusion The result is a set of four rationally designed vectors, which can be used for streamlined cloning, expression and purification of target proteins in the laboratory and have the potential for being adaptable to a high

  6. Activation of human natural killer cells by the soluble form of cellular prion protein

    Energy Technology Data Exchange (ETDEWEB)

    Seong, Yeon-Jae [Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of); Hafis Clinic, Seoul (Korea, Republic of); Sung, Pil Soo; Jang, Young-Soon; Choi, Young Joon [Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of); Park, Bum-Chan [Aging Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon (Korea, Republic of); Park, Su-Hyung [Laboratory of Translational Immunology and Vaccinology, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of); Park, Young Woo [Aging Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon (Korea, Republic of); Shin, Eui-Cheol, E-mail: ecshin@kaist.ac.kr [Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of)

    2015-08-21

    Cellular prion protein (PrP{sup C}) is widely expressed in various cell types, including cells of the immune system. However, the specific roles of PrP{sup C} in the immune system have not been clearly elucidated. In the present study, we investigated the effects of a soluble form of recombinant PrP{sup C} protein on human natural killer (NK) cells. Recombinant soluble PrP{sup C} protein was generated by fusion of human PrP{sup C} with the Fc portion of human IgG{sub 1} (PrP{sup C}-Fc). PrP{sup C}-Fc binds to the surface of human NK cells, particularly to CD56{sup dim} NK cells. PrP{sup C}-Fc induced the production of cytokines and chemokines and the degranulation of granzyme B from NK cells. In addition, PrP{sup C}-Fc facilitated the IL-15-induced proliferation of NK cells. PrP{sup C}-Fc induced phosphorylation of ERK-1/2 and JNK in NK cells, and inhibitors of the ERK or the JNK pathways abrogated PrP{sup C}-Fc-induced cytokine production in NK cells. In conclusion, the soluble form of recombinant PrP{sup C}-Fc protein activates human NK cells via the ERK and JNK signaling pathways. - Highlights: • Recombinant soluble PrP{sup C} (PrP{sup C}-Fc) was generated by fusion of human PrP{sup C} with IgG1 Fc portion. • PrP{sup C}-Fc protein induces the production of cytokines and degranulation from human NK cells. • PrP{sup C}-Fc protein enhances the IL-15-induced proliferation of human NK cells. • PrP{sup C}-Fc protein activates human NK cells via the ERK and JNK signaling pathways.

  7. Soluble cysteine-rich tick saliva proteins Salp15 and Iric-1 from E. coli.

    Science.gov (United States)

    Kolb, Philipp; Vorreiter, Jolanta; Habicht, Jüri; Bentrop, Detlef; Wallich, Reinhard; Nassal, Michael

    2015-01-01

    Ticks transmit numerous pathogens, including borreliae, which cause Lyme disease. Tick saliva contains a complex mix of anti-host defense factors, including the immunosuppressive cysteine-rich secretory glycoprotein Salp15 from Ixodes scapularis ticks and orthologs like Iric-1 from Ixodes ricinus. All tick-borne microbes benefit from the immunosuppression at the tick bite site; in addition, borreliae exploit the binding of Salp15 to their outer surface protein C (OspC) for enhanced transmission. Hence, Salp15 proteins are attractive targets for anti-tick vaccines that also target borreliae. However, recombinant Salp proteins are not accessible in sufficient quantity for either vaccine manufacturing or for structural characterization. As an alternative to low-yield eukaryotic systems, we investigated cytoplasmic expression in Escherichia coli, even though this would not result in glycosylation. His-tagged Salp15 was efficiently expressed but insoluble. Among the various solubility-enhancing protein tags tested, DsbA was superior, yielding milligram amounts of soluble, monomeric Salp15 and Iric-1 fusions. Easily accessible mutants enabled epitope mapping of two monoclonal antibodies that, importantly, cross-react with glycosylated Salp15, and revealed interaction sites with OspC. Free Salp15 and Iric-1 from protease-cleavable fusions, despite limited solubility, allowed the recording of (1)H-(15)N 2D NMR spectra, suggesting partial folding of the wild-type proteins but not of Cys-free variants. Fusion to the NMR-compatible GB1 domain sufficiently enhanced solubility to reveal first secondary structure elements in (13)C/(15)N double-labeled Iric-1. Together, E. coli expression of appropriately fused Salp15 proteins may be highly valuable for the molecular characterization of the function and eventually the 3D structure of these medically relevant tick proteins.

  8. How to find soluble proteins: a comprehensive analysis of alpha/beta hydrolases for recombinant expression in E. coli

    Directory of Open Access Journals (Sweden)

    Barth Sandra

    2005-04-01

    Full Text Available Abstract Background In screening of libraries derived by expression cloning, expression of active proteins in E. coli can be limited by formation of inclusion bodies. In these cases it would be desirable to enrich gene libraries for coding sequences with soluble gene products in E. coli and thus to improve the efficiency of screening. Previously Wilkinson and Harrison showed that solubility can be predicted from amino acid composition (Biotechnology 1991, 9(5:443–448. We have applied this analysis to members of the alpha/beta hydrolase fold family to predict their solubility in E. coli. alpha/beta hydrolases are a highly diverse family with more than 1800 proteins which have been grouped into homologous families and superfamilies. Results The predicted solubility in E. coli depends on hydrolase size, phylogenetic origin of the host organism, the homologous family and the superfamily, to which the hydrolase belongs. In general small hydrolases are predicted to be more soluble than large hydrolases, and eukaryotic hydrolases are predicted to be less soluble in E. coli than prokaryotic ones. However, combining phylogenetic origin and size leads to more complex conclusions. Hydrolases from prokaryotic, fungal and metazoan origin are predicted to be most soluble if they are of small, medium and large size, respectively. We observed large variations of predicted solubility between hydrolases from different homologous families and from different taxa. Conclusion A comprehensive analysis of all alpha/beta hydrolase sequences allows more efficient screenings for new soluble alpha/beta hydrolases by the use of libraries which contain more soluble gene products. Screening of hydrolases from families whose members are hard to express as soluble proteins in E. coli should first be done in coding sequences of organisms from phylogenetic groups with the highest average of predicted solubility for proteins of this family. The tools developed here can be used

  9. CVs and millisecond pulsar progenitors in globular clusters

    Science.gov (United States)

    Grindlay, J. E.; Cool, A. M.; Bailyn, C. D.

    1991-01-01

    The recent discovery of a large population of millisecond pulsars in globular clusters, together with earlier studies of both low luminosity X-ray sources and LMXBs in globulars, suggest there should be significant numbers of CVs in globulars. Although they have been searched for without success in selected cluster X-ray source fields, systematic surveys are lacking and would constrain binary production and both stellar and dynamical evolution in globular clusters. We describe the beginnings of such a search, using narrow band H-alpha imaging, and the sensitivities it might achieve.

  10. Globular Clusters Shine in a Galaxy Lacking Dark Matter

    Science.gov (United States)

    Kohler, Susanna

    2018-04-01

    You may have seen recent news about NGC 1052DF2, a galaxy that was discovered to have little or no dark matter. Now, a new study explores what NGC 1052DF2 does have: an enigmatic population of unusually large and luminous globular clusters.Keck/LRIS spectra (left and right) and HST images (center) of the 11 clusters associated with NGC 1052DF2. The color images each span 1 1. [van Dokkum et al. 2018]An Unusual DwarfThe ultra-diffuse galaxy NGC 1052DF2, originally identified with the Dragonfly Telescope Array, has puzzled astronomers since the discovery that its dynamical mass determined by the motions of globular-cluster-like objects spotted within it is essentially the same as its stellar mass. This equivalence implies that the galaxy is strangely lacking dark matter; the upper limit set on its dark matter halo is 400 times smaller than what we would expect for such a dwarf galaxy.Led by Pieter van Dokkum (Yale University), the team that made this discovery has now followed up with detailed Hubble Space Telescope imaging and Keck spectroscopy. Their goal? To explore the objects that allowed them to make the dynamical-mass measurement: the oddly bright globular clusters of NGC 1052DF2.Sizes (circularized half-light radii) vs. absolute magnitudes for globular clusters in NGC1052DF2 (black) and the Milky Way (red). [Adapted from van Dokkum et al. 2018]Whats Up with the Globular Clusters?Van Dokkum and collaborators spectroscopically confirmed 11 compact objects associated with the faint galaxy. These objects are globular-cluster-like in their appearance, but the peak of their luminosity distribution is offset by a factor of four from globular clusters of other galaxies; these globular clusters are significantly brighter than is typical.Using the Hubble imaging, the authors determined that NGC 1052DF2s globular clusters are more than twice the size of the Milky Ways globular clusters in the same luminosity range. As is typical for globular clusters, they are an old

  11. Prokaryotic Soluble Overexpression and Purification of Human VEGF165 by Fusion to a Maltose Binding Protein Tag.

    Directory of Open Access Journals (Sweden)

    Minh Tan Nguyen

    Full Text Available Human vascular endothelial growth factor (VEGF is a key regulator of angiogenesis and plays a central role in the process of tumor growth and metastatic dissemination. Escherichia coli is one of the most common expression systems used for the production of recombinant proteins; however, expression of human VEGF in E. coli has proven difficult because the E. coli-expressed VEGF tends to be misfolded and forms inclusion bodies, resulting in poor solubility. In this study, we successfully produced semi-preparative amounts of soluble bioactive human VEGF165 (hVEGF. We created seven N-terminal fusion tag constructs with hexahistidine (His6, thioredoxin (Trx, glutathione S-transferase (GST, maltose-binding protein (MBP, N-utilization substance protein A (NusA, human protein disulfide isomerase (PDI, and the b'a' domain of PDI (PDIb'a', and tested each construct for soluble overexpression in E. coli. We found that at 18°C, 92.8% of the MBP-tagged hVEGF to be soluble and that this tag significantly increased the protein's solubility. We successfully purified 0.8 mg of pure hVEGF per 500 mL cell culture. The purified hVEGF is stable after tag cleavage, contains very low levels of endotoxin, and is 97.6% pure. Using an Flk1+ mesodermal precursor cell (MPC differentiation assay, we show that the purified hVEGF is not only bioactive but has similar bioactivity to hVEGF produced in mammalian cells. Previous reports on producing hVEGF in E. coli have all been based on refolding of the protein from inclusion bodies. To our knowledge, this is the first report on successfully expressing and purifying soluble hVEGF in E. coli.

  12. Photodamaging mechanism of the eye structure: UV effect on soluble proteins of the lens

    International Nuclear Information System (INIS)

    Korkhmazyan, M.M.; Fedorovich, I.B.; Ostrovskij, M.A.

    1983-01-01

    Damaging effect of UV-radiation on soluble proteins of bull lens has been studied. Irradiation results in lens proteins growing yellow, new absorption bands with the maxima 245 and 305 nm appear. It is shown that during photodamage oxidation of SH-groups takes place and protein aggregates are formed

  13. Biological assessment of neonicotinoids imidacloprid and its major metabolites for potentially human health using globular proteins as a model.

    Science.gov (United States)

    Ding, Fei; Peng, Wei

    2015-06-01

    The assessment of biological activities of imidacloprid and its two major metabolites, namely 6-chloronicotinic acid and 2-imidazolidone for nontarget organism, by employing essentially functional biomacromolecules, albumin and hemoglobin as a potentially model with the use of circular dichroism (CD), fluorescence, extrinsic 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence as well as molecular modeling is the theme of this work. By dint of CD spectra and synchronous fluorescence, it was clear that the orderly weak interactions between amino acid residues within globular proteins were disturbed by imidacloprid, and this event led to marginally alterations or self-regulations of protein conformation so as to lodge imidacloprid more tightly. Both steady state and time-resolved fluorescence suggested that the fluorescence of Trp residues in proteins was quenched after the presence of imidacloprid, corresponding to noncovalent protein-imidacloprid complexes formation and, the reaction belongs to moderate association (K=1.888/1.614×10(4)M(-1) for albumin/hemoglobin-imidacloprid, respectively), hydrogen bonds and π stacking performed a vital role in stabilizing the complexes, as derived from thermodynamic analysis and molecular modeling. With the aid of hydrophobic ANS experiments, subdomain IIA and α1β2 interface of albumin and hemoglobin, respectively, were found to be preserved high-affinity for imidacloprid. These results ties in with the subsequently molecular modeling laying imidacloprid in the Sudlow's site I and close to Trp-213 residue on albumin, while settling down B/Trp-37 residue nearby in hemoglobin, and these conclusions further confirmed by site-directed mutagenesis and molecular dynamics simulation. But, at the same time, several crucial noncovalent bonds came from other amino acid residues, e.g. Arg-194 and Arg-198 (albumin) and B/Arg-40, B/Asp-99 and B/Asn-102 (hemoglobin) cannot be ignored completely. Based on the comparative studies of

  14. Benzoate-mediated changes on expression profile of soluble proteins in Serratia sp. DS001.

    Science.gov (United States)

    Pandeeti, E V P; Chinnaboina, M R; Siddavattam, D

    2009-05-01

    To assess differences in protein expression profile associated with shift in carbon source from succinate to benzoate in Serratia sp. DS001 using a proteomics approach. A basic proteome map was generated for the soluble proteins extracted from Serratia sp. DS001 grown in succinate and benzoate. The differently and differentially expressed proteins were identified using ImageMaster 2D Platinum software (GE Healthcare). The identity of the proteins was determined by employing MS or MS/MS. Important enzymes such as Catechol 1,2 dioxygenase and transcriptional regulators that belong to the LysR superfamily were identified. Nearly 70 proteins were found to be differentially expressed when benzoate was used as carbon source. Based on the protein identity and degradation products generated from benzoate it is found that ortho pathway is operational in Serratia sp. DS001. Expression profile of the soluble proteins associated with shift in carbon source was mapped. The study also elucidates degradation pathway of benzoate in Serratia sp. DS001 by correlating the proteomics data with the catabolites of benzoate.

  15. Properties of spores of Bacillus subtilis strains which lack the major small, acid-soluble protein

    International Nuclear Information System (INIS)

    Hackett, R.H.; Setlow, P.

    1988-01-01

    Bacillus subtilis strains containing a deletion in the gene coding for the major small, acid-soluble, spore protein (SASP-gamma) grew and sporulated, and their spores initiated germination normally, but outgrowth of SASP-gamma- spores was significantly slower than that of wild-type spores. The absence of SASP-gamma had no effect on spore protoplast density or spore resistance to heat or radiation. Consequently, SASP-gamma has a different function in spores than do the other major small, acid-soluble proteins

  16. Soluble lens protein polymorphism in flying fishes from the central Arabian Sea

    Digital Repository Service at National Institute of Oceanography (India)

    Menezes, M.R.

    Soluble eye lens nuclei proteins of flying-fishes were studied by cellogel electrophoresis. Three distinct patterns characterized by the number of bands, mobility and staining intensity were observed. Morphological studies of these fishes showed...

  17. Prefoldin Promotes Proteasomal Degradation of Cytosolic Proteins with Missense Mutations by Maintaining Substrate Solubility.

    Directory of Open Access Journals (Sweden)

    Sophie A Comyn

    2016-07-01

    Full Text Available Misfolded proteins challenge the ability of cells to maintain protein homeostasis and can accumulate into toxic protein aggregates. As a consequence, cells have adopted a number of protein quality control pathways to prevent protein aggregation, promote protein folding, and target terminally misfolded proteins for degradation. In this study, we employed a thermosensitive allele of the yeast Guk1 guanylate kinase as a model misfolded protein to investigate degradative protein quality control pathways. We performed a flow cytometry based screen to identify factors that promote proteasomal degradation of proteins misfolded as the result of missense mutations. In addition to the E3 ubiquitin ligase Ubr1, we identified the prefoldin chaperone subunit Gim3 as an important quality control factor. Whereas the absence of GIM3 did not impair proteasomal function or the ubiquitination of the model substrate, it led to the accumulation of the poorly soluble model substrate in cellular inclusions that was accompanied by delayed degradation. We found that Gim3 interacted with the Guk1 mutant allele and propose that prefoldin promotes the degradation of the unstable model substrate by maintaining the solubility of the misfolded protein. We also demonstrated that in addition to the Guk1 mutant, prefoldin can stabilize other misfolded cytosolic proteins containing missense mutations.

  18. Prokaryotic soluble overexpression and purification of bioactive human growth hormone by fusion to thioredoxin, maltose binding protein, and protein disulfide isomerase.

    Directory of Open Access Journals (Sweden)

    Minh Tan Nguyen

    Full Text Available Human growth hormone (hGH is synthesized by somatotroph cells of the anterior pituitary gland and induces cell proliferation and growth. This protein has been approved for the treatment of various conditions, including hGH deficiency, chronic renal failure, and Turner syndrome. Efficient production of hGH in Escherichia coli (E. coli has proven difficult because the E. coli-expressed hormone tends to aggregate and form inclusion bodies, resulting in poor solubility. In this study, seven N-terminal fusion partners, hexahistidine (His6, thioredoxin (Trx, glutathione S-transferase (GST, maltose-binding protein (MBP, N-utilization substance protein A (NusA, protein disulfide bond isomerase (PDI, and the b'a' domain of PDI (PDIb'a', were tested for soluble overexpression of codon-optimized hGH in E. coli. We found that MBP and hPDI tags significantly increased the solubility of the hormone. In addition, lowering the expression temperature to 18°C also dramatically increased the solubility of all the fusion proteins. We purified hGH from MBP-, PDIb'a'-, or Trx-tagged hGH expressed at 18°C in E. coli using simple chromatographic techniques and compared the final purity, yield, and activity of hGH to assess the impact of each partner protein. Purified hGH was highly pure on silver-stained gel and contained very low levels of endotoxin. On average, ∼37 mg, ∼12 mg, and ∼7 mg of hGH were obtained from 500 mL-cell cultures of Trx-hGH, MBP-hGH, and PDIb'a'-hGH, respectively. Subsequently, hGH was analyzed using mass spectroscopy to confirm the presence of two intra-molecular disulfide bonds. The bioactivity of purified hGHs was demonstrated using Nb2-11 cell.

  19. ESPRIT: an automated, library-based method for mapping and soluble expression of protein domains from challenging targets.

    Science.gov (United States)

    Yumerefendi, Hayretin; Tarendeau, Franck; Mas, Philippe J; Hart, Darren J

    2010-10-01

    Expression of sufficient quantities of soluble protein for structural biology and other applications is often a very difficult task, especially when multimilligram quantities are required. In order to improve yield, solubility or crystallisability of a protein, it is common to subclone shorter genetic constructs corresponding to single- or multi-domain fragments. However, it is not always clear where domain boundaries are located, especially when working on novel targets with little or no sequence similarity to other proteins. Several methods have been described employing aspects of directed evolution to the recombinant expression of challenging proteins. These combine the construction of a random library of genetic constructs of a target with a screening or selection process to identify solubly expressing protein fragments. Here we review several datasets from the ESPRIT (Expression of Soluble Proteins by Random Incremental Truncation) technology to provide a view on its capabilities. Firstly, we demonstrate how it functions using the well-characterised NF-kappaB p50 transcription factor as a model system. Secondly, application of ESPRIT to the challenging PB2 subunit of influenza polymerase has led to several novel atomic resolution structures; here we present an overview of the screening phase of that project. Thirdly, analysis of the human kinase TBK1 is presented to show how the ESPRIT technology rapidly addresses the compatibility of challenging targets with the Escherichia coli expression system.

  20. Synergistic enhancement in the co-gelation of salt-soluble pea proteins and whey proteins.

    Science.gov (United States)

    Wong, Douglas; Vasanthan, Thava; Ozimek, Lech

    2013-12-15

    This paper investigated the enhancement of thermal gelation properties when salt-soluble pea proteins were co-gelated with whey proteins in NaCl solutions, using different blend ratios, total protein concentrations, pH, and salt concentrations. Results showed that the thermal co-gelation of pea/whey proteins blended in ratio of 2:8 in NaCl solutions showed synergistic enhancement in storage modulus, gel hardness, paste viscosity and minimum gelation concentrations. The highest synergistic enhancement was observed at pH 6.0 as compared with pH 4.0 and 8.0, and at the lower total protein concentration of 10% as compared with 16% and 22% (w/v), as well as in lower NaCl concentrations of 0.5% and 1.0% as compared with 1.5%, 2.0%, 2.5%, and 3.0% (w/v). The least gelation concentrations were also lower in the different pea/whey protein blend ratios than in pure pea or whey proteins, when dissolved in 1.0% or 2.5% (w/v) NaCl aqueous solutions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. Reconstructing galaxy histories from globular clusters.

    Science.gov (United States)

    West, Michael J; Côté, Patrick; Marzke, Ronald O; Jordán, Andrés

    2004-01-01

    Nearly a century after the true nature of galaxies as distant 'island universes' was established, their origin and evolution remain great unsolved problems of modern astrophysics. One of the most promising ways to investigate galaxy formation is to study the ubiquitous globular star clusters that surround most galaxies. Globular clusters are compact groups of up to a few million stars. They generally formed early in the history of the Universe, but have survived the interactions and mergers that alter substantially their parent galaxies. Recent advances in our understanding of the globular cluster systems of the Milky Way and other galaxies point to a complex picture of galaxy genesis driven by cannibalism, collisions, bursts of star formation and other tumultuous events.

  2. Nova-driven winds in globular clusters

    International Nuclear Information System (INIS)

    Scott, E.H.; Durisen, R.H.

    1978-01-01

    Recent sensitive searches for Hα emission from ionized intracluster gas in globular clusters have set upper limits that conflict with theoretical predictions. We suggest that nova outbursts heat the gas, producing winds that resolve this discrepancy. The incidence of novae in globular clusters, the conversion of kinetic energy of the nova shell to thermal energy of the intracluster gas, and the characteristics of the resultant winds are discussed. Calculated emission from the nova-driven models does not conflict with any observations to date. Some suggestions are made concerning the most promising approaches for future detection of intracluster gas on the basis of these models. The possible relationship of nova-driven winds of globular cluster X-ray sources is also considered

  3. Shaping Globular Clusters with Black Holes

    Science.gov (United States)

    Kohler, Susanna

    2018-03-01

    How many black holes lurk within the dense environments of globular clusters, and how do these powerful objects shape the properties of the cluster around them? One such cluster, NGC 3201, is now helping us to answer these questions.Hunting Stellar-Mass Black HolesSince the detection of merging black-hole binaries by the Laser Interferometer Gravitational-Wave Observatory (LIGO), the dense environments of globular clusters have received increasing attention as potential birthplaces of these compact binary systems.The central region of the globular star cluster NGC 3201, as viewed by Hubble. The black hole is in orbit with the star marked by the blue circle. [NASA/ESA]In addition, more and more stellar-mass black-hole candidates have been observed within globular clusters, lurking in binary pairs with luminous, non-compact companions. The most recent of these detections, found in the globular cluster NGC 3201, stands alone as the first stellar-mass black hole candidate discovered via radial velocity observations: the black holes main-sequence companion gave away its presence via a telltale wobble.Now a team of scientists led by Kyle Kremer (CIERA and Northwestern University) is using models of this system to better understand the impact that black holes might have on their host clusters.A Model ClusterThe relationship between black holes and their host clusters is complicated. Though the cluster environment can determine the dynamical evolution of the black holes, the retention rate of black holes in a globular cluster (i.e., how many remain in the cluster when they are born as supernovae, rather than being kicked out during the explosion) influences how the host cluster evolves.Kremer and collaborators track this complex relationship by modeling the evolution of a cluster similar to NGC 3201 with a Monte Carlo code. The code incorporates physics relevant to the evolution of black holes and black-hole binaries in globular clusters, such as two-body relaxation

  4. Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli

    DEFF Research Database (Denmark)

    Sørensen, Hans; Mortensen, Kim

    2005-01-01

    Pure, soluble and functional proteins are of high demand in modern biotechnology. Natural protein sources rarely meet the requirements for quantity, ease of isolation or price and hence recombinant technology is often the method of choice. Recombinant cell factories are constantly employed...... molecular tools available. In spite of all these qualities, expression of recombinant proteins with E. coli as the host often results in insoluble and/or nonfunctional proteins. Here we review new approaches to overcome these obstacles by strategies that focus on either controlled expression of target...... for the production of protein preparations bound for downstream purification and processing. Eschericia coli is a frequently used host, since it facilitates protein expression by its relative simplicity, its inexpensive and fast high density cultivation, the well known genetics and the large number of compatible...

  5. Dinamical properties of globular clusters: Primordial or evolutional?

    Science.gov (United States)

    Surdin, V. G.

    1995-04-01

    Some observable relations between globular cluster parameters appear as a result of dynamical evolution of the cluster system. These relations are inapplicable to the studies of the globular cluster origin

  6. GLOBULAR CLUSTERS AS CRADLES OF LIFE AND ADVANCED CIVILIZATIONS

    Energy Technology Data Exchange (ETDEWEB)

    Stefano, R. Di [Harvard-Smithsonian Center for Astrophysics (United States); Ray, A., E-mail: rdistefano@cfa.harvard.edu, E-mail: akr@tifr.res.in [Tata Institute of Fundamental Research (India)

    2016-08-10

    Globular clusters are ancient stellar populations in compact dense ellipsoids. There is no star formation and there are no core-collapse supernovae, but several lines of evidence suggest that globular clusters are rich in planets. If so, and if advanced civilizations can develop there, then the distances between these civilizations and other stars would be far smaller than typical distances between stars in the Galactic disk, facilitating interstellar communication and travel. The potent combination of long-term stability and high stellar densities provides a globular cluster opportunity. Yet the very proximity that promotes interstellar travel also brings danger, as stellar interactions can destroy planetary systems. We find, however, that large portions of many globular clusters are “sweet spots,” where habitable-zone planetary orbits are stable for long times. Globular clusters in our own and other galaxies are, therefore, among the best targets for searches for extraterrestrial intelligence (SETI). We use the Drake equation to compare the likelihood of advanced civilizations in globular clusters to that in the Galactic disk. We also consider free-floating planets, since wide-orbit planets can be ejected to travel through the cluster. Civilizations spawned in globular clusters may be able to establish self-sustaining outposts, reducing the probability that a single catastrophic event will destroy the civilization. Although individual civilizations may follow different evolutionary paths, or even be destroyed, the cluster may continue to host advanced civilizations once a small number have jumped across interstellar space. Civilizations residing in globular clusters could therefore, in a sense, be immortal.

  7. GLOBULAR CLUSTERS AS CRADLES OF LIFE AND ADVANCED CIVILIZATIONS

    International Nuclear Information System (INIS)

    Stefano, R. Di; Ray, A.

    2016-01-01

    Globular clusters are ancient stellar populations in compact dense ellipsoids. There is no star formation and there are no core-collapse supernovae, but several lines of evidence suggest that globular clusters are rich in planets. If so, and if advanced civilizations can develop there, then the distances between these civilizations and other stars would be far smaller than typical distances between stars in the Galactic disk, facilitating interstellar communication and travel. The potent combination of long-term stability and high stellar densities provides a globular cluster opportunity. Yet the very proximity that promotes interstellar travel also brings danger, as stellar interactions can destroy planetary systems. We find, however, that large portions of many globular clusters are “sweet spots,” where habitable-zone planetary orbits are stable for long times. Globular clusters in our own and other galaxies are, therefore, among the best targets for searches for extraterrestrial intelligence (SETI). We use the Drake equation to compare the likelihood of advanced civilizations in globular clusters to that in the Galactic disk. We also consider free-floating planets, since wide-orbit planets can be ejected to travel through the cluster. Civilizations spawned in globular clusters may be able to establish self-sustaining outposts, reducing the probability that a single catastrophic event will destroy the civilization. Although individual civilizations may follow different evolutionary paths, or even be destroyed, the cluster may continue to host advanced civilizations once a small number have jumped across interstellar space. Civilizations residing in globular clusters could therefore, in a sense, be immortal.

  8. An autoclave treatment reduces the solubility and antigenicity of an allergenic protein found in buckwheat flour.

    Science.gov (United States)

    Tomotake, Hiroyuki; Yamazaki, Rikio; Yamato, Masayuki

    2012-06-01

    The effects of an autoclave treatment of buckwheat flour on a 24-kDa allergenic protein were investigated by measuring reduction in solubility and antibody binding. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the intensity of the major bands, including that of the 24-kDa allergen, was reduced by the autoclave treatment. The protein solubility in buckwheat flour was variably decreased by the autoclave treatment. Enzyme-linked immunosorbent assay analysis using a monoclonal antibody specific for buckwheat 24-kDa protein showed that the reactivity of protein extracts (10 μg/ml) from buckwheat flour was lowered by the autoclave treatment. The autoclave treatment may reduce the major allergen content of buckwheat. Future studies will determine if autoclaving treatments affect the allergenicity of the 24-kDa buckwheat protein.

  9. Search for optical millisecond pulsars in globular clusters

    International Nuclear Information System (INIS)

    Middleditch, J.H.; Imamura, J.N.; Steiman-Cameron, T.Y.

    1988-01-01

    A search for millisecond optical pulsars in several bright, compact globular clusters was conducted. The sample included M28, and the X-ray clusters 47 Tuc, NGC 6441, NGC 6624, M22, and M15. The globular cluster M28 contains the recently discovered 327 Hz radio pulsar. Upper limits of 4 sigma to pulsed emission of (1-20) solar luminosities were found for the globular clusters tested, and 0.3 solar luminosity for the M28 pulsar for frequencies up to 500 Hz. 8 references

  10. Transport of soluble proteins through the Golgi occurs by diffusion via continuities across cisternae

    Science.gov (United States)

    Beznoussenko, Galina V; Parashuraman, Seetharaman; Rizzo, Riccardo; Polishchuk, Roman; Martella, Oliviano; Di Giandomenico, Daniele; Fusella, Aurora; Spaar, Alexander; Sallese, Michele; Capestrano, Maria Grazia; Pavelka, Margit; Vos, Matthijn R; Rikers, Yuri GM; Helms, Volkhard; Mironov, Alexandre A; Luini, Alberto

    2014-01-01

    The mechanism of transport through the Golgi complex is not completely understood, insofar as no single transport mechanism appears to account for all of the observations. Here, we compare the transport of soluble secretory proteins (albumin and α1-antitrypsin) with that of supramolecular cargoes (e.g., procollagen) that are proposed to traverse the Golgi by compartment progression–maturation. We show that these soluble proteins traverse the Golgi much faster than procollagen while moving through the same stack. Moreover, we present kinetic and morphological observations that indicate that albumin transport occurs by diffusion via intercisternal continuities. These data provide evidence for a transport mechanism that applies to a major class of secretory proteins and indicate the co-existence of multiple intra-Golgi trafficking modes. DOI: http://dx.doi.org/10.7554/eLife.02009.001 PMID:24867214

  11. New VVV Survey Globular Cluster Candidates in the Milky Way Bulge

    Energy Technology Data Exchange (ETDEWEB)

    Minniti, Dante; Gómez, Matías [Departamento de Física, Facultad de Ciencias Exactas, Universidad Andrés Bello, Av. Fernandez Concha 700, Las Condes, Santiago (Chile); Geisler, Douglas; Fernández-Trincado, Jose G. [Departamento de Astronomía, Casilla 160-C, Universidad de Concepción, Casilla 160-C, Concepción (Chile); Alonso-García, Javier; Beamín, Juan Carlos; Borissova, Jura; Catelan, Marcio; Ramos, Rodrigo Contreras; Kurtev, Radostin; Pullen, Joyce [Instituto Milenio de Astrofísica, Santiago (Chile); Palma, Tali; Clariá, Juan J. [Observatorio Astronómico, Universidad Nacional de Córdoba, Laprida 854, Córdoba (Argentina); Cohen, Roger E. [Space Telescope Science Institute, 2700 San Martin Drive, Baltimore (United States); Dias, Bruno [European Southern Observatory, Alonso de Cordova 3107, Vitacura, Santiago (Chile); Hempel, Maren [Pontificia Universidad Católica de Chile, Instituto de Astrofísica, Av. Vicuña Mackenna 4860, Santiago (Chile); Ivanov, Valentin D. [European Southern Observatory, Karl-Schwarszchild-Str. 2, D-85748 Garching bei Muenchen (Germany); Lucas, Phillip W. [Dept. of Astronomy, University of Hertfordshire, Hertfordshire (United Kingdom); Moni-Bidin, Christian; Alegría, Sebastian Ramírez [Instituto de Astronomía, Universidad Católica del Norte, Antofagasta (Chile); and others

    2017-11-10

    It is likely that a number of Galactic globular clusters remain to be discovered, especially toward the Galactic bulge. High stellar density combined with high and differential interstellar reddening are the two major problems for finding globular clusters located toward the bulge. We use the deep near-IR photometry of the VISTA Variables in the Vía Láctea (VVV) Survey to search for globular clusters projected toward the Galactic bulge, and hereby report the discovery of 22 new candidate globular clusters. These objects, detected as high density regions in our maps of bulge red giants, are confirmed as globular cluster candidates by their color–magnitude diagrams. We provide their coordinates as well as their near-IR color–magnitude diagrams, from which some basic parameters are derived, such as reddenings and heliocentric distances. The color–magnitude diagrams reveal well defined red giant branches in all cases, often including a prominent red clump. The new globular cluster candidates exhibit a variety of extinctions (0.06 < A {sub Ks} < 2.77) and distances (5.3 < D < 9.5 kpc). We also classify the globular cluster candidates into 10 metal-poor and 12 metal-rich clusters, based on the comparison of their color–magnitude diagrams with those of known globular clusters also observed by the VVV Survey. Finally, we argue that the census for Galactic globular clusters still remains incomplete, and that many more candidate globular clusters (particularly the low luminosity ones) await to be found and studied in detail in the central regions of the Milky Way.

  12. Photometric studies of globular clusters in the Andromeda nebula

    International Nuclear Information System (INIS)

    Sharov, A.S.; Lyutyj, V.M.

    1983-01-01

    The comparison of the frequency distribution of Bergh Q and Racine R metallicity parameters for globular clusters in the Galaxy and M31 is given. Mean values of the parameters are: in the Galaxy anti Q=-0.31 and anti R=0.40, in M31 anti Q=-0.32 and anti R=0.42. Hence the mean metallicity of globular clusters in two galaxies is identical. The differences in the observed frequency distribution of the parameters, in particular in the limits of general metallicity, are related to the random errors of photometrical measurements of globular clusters, considerably greater in the case of M31. Thereby the preference should be given to Hanes conclusion that globular clusters form a uniform population at least in two close systems. It should not be excepted that in other galaxies mean colour characteristics and hence metallicity of clusters may be of other type. Thus globular clusters related to the M31-NGC 205 satellite have somewhat minor B-V colour factors

  13. Functionality of extrusion--texturized whey proteins.

    Science.gov (United States)

    Onwulata, C I; Konstance, R P; Cooke, P H; Farrell, H M

    2003-11-01

    Whey, a byproduct of the cheesemaking process, is concentrated by processors to make whey protein concentrates (WPC) and isolates (WPI). Only 50% of whey proteins are used in foods. In order to increase their usage, texturizing WPC, WPI, and whey albumin is proposed to create ingredients with new functionality. Extrusion processing texturizes globular proteins by shearing and stretching them into aligned or entangled fibrous bundles. In this study, WPC, WPI, and whey albumin were extruded in a twin screw extruder at approximately 38% moisture content (15.2 ml/min, feed rate 25 g/min) and, at different extrusion cook temperatures, at the same temperature for the last four zones before the die (35, 50, 75, and 100 degrees C, respectively). Protein solubility, gelation, foaming, and digestibility were determined in extrudates. Degree of extrusion-induced insolubility (denaturation) or texturization, determined by lack of solubility at pH 7 for WPI, increased from 30 to 60, 85, and 95% for the four temperature conditions 35, 50, 75, and 100 degrees C, respectively. Gel strength of extruded isolates increased initially 115% (35 degrees C) and 145% (50 degrees C), but gel strength was lost at 75 and 100 degrees C. Denaturation at these melt temperatures had minimal effect on foaming and digestibility. Varying extrusion cook temperature allowed a new controlled rate of denaturation, indicating that a texturized ingredient with a predetermined functionality based on degree of denaturation can be created.

  14. The interactions between CdTe quantum dots and proteins: understanding nano-bio interface

    Directory of Open Access Journals (Sweden)

    Shreeram S. Joglekar

    2017-01-01

    Full Text Available Despite remarkable developments in the nanoscience, relatively little is known about the physical (electrostatic interactions of nanoparticles with bio macromolecules. These interactions can influence the properties of both nanoparticles and the bio-macromolecules. Understanding this bio-interface is a prerequisite to utilize both nanoparticles and biomolecules for bioengineering. In this study, luminescent, water soluble CdTe quantum dots (QDs capped with mercaptopropionic acid (MPA were synthesized by organometallic method and then interaction between nanoparticles (QDs and three different types of proteins (BSA, Lysozyme and Hemoglobin were investigated by fluorescence spectroscopy at pH= 7.4. Based on fluorescence quenching results, Stern-Volmer quenching constant (Ksv, binding constant (Kq and binding sites (n for proteins were calculated. The results show that protein structure (e.g.,globular, metalloprotein, etc. has a significant role in Protein-Quantum dots interactions and each type of protein influence physicochemical properties of Quantum dots differently.

  15. TOWARDS THE CONCEPT OF GLOBULAR PEARLITE

    OpenAIRE

    A. G. Anisovich; M. K. Stepankova; A. A. Andrushevich

    2016-01-01

    Explanatory imprecisions of concept of globular pearlite and ferrite-carbide-mixture are considered. The need of concept binding of globular pearlite to specific grain with 0.8% carbon content is explained with the assistance of exemplary data obtained at the present metallographic equipment. The question of educational material presentation concerning the process of teaching of discipline «Materials and construction materials technology» is discussed in relation to the educational process of...

  16. High-throughput method for optimum solubility screening for homogeneity and crystallization of proteins

    Science.gov (United States)

    Kim, Sung-Hou [Moraga, CA; Kim, Rosalind [Moraga, CA; Jancarik, Jamila [Walnut Creek, CA

    2012-01-31

    An optimum solubility screen in which a panel of buffers and many additives are provided in order to obtain the most homogeneous and monodisperse protein condition for protein crystallization. The present methods are useful for proteins that aggregate and cannot be concentrated prior to setting up crystallization screens. A high-throughput method using the hanging-drop method and vapor diffusion equilibrium and a panel of twenty-four buffers is further provided. Using the present methods, 14 poorly behaving proteins have been screened, resulting in 11 of the proteins having highly improved dynamic light scattering results allowing concentration of the proteins, and 9 were crystallized.

  17. Foldability of a Natural De Novo Evolved Protein.

    Science.gov (United States)

    Bungard, Dixie; Copple, Jacob S; Yan, Jing; Chhun, Jimmy J; Kumirov, Vlad K; Foy, Scott G; Masel, Joanna; Wysocki, Vicki H; Cordes, Matthew H J

    2017-11-07

    The de novo evolution of protein-coding genes from noncoding DNA is emerging as a source of molecular innovation in biology. Studies of random sequence libraries, however, suggest that young de novo proteins will not fold into compact, specific structures typical of native globular proteins. Here we show that Bsc4, a functional, natural de novo protein encoded by a gene that evolved recently from noncoding DNA in the yeast S. cerevisiae, folds to a partially specific three-dimensional structure. Bsc4 forms soluble, compact oligomers with high β sheet content and a hydrophobic core, and undergoes cooperative, reversible denaturation. Bsc4 lacks a specific quaternary state, however, existing instead as a continuous distribution of oligomer sizes, and binds dyes indicative of amyloid oligomers or molten globules. The combination of native-like and non-native-like properties suggests a rudimentary fold that could potentially act as a functional intermediate in the emergence of new folded proteins de novo. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. In meso in situ serial X-ray crystallography of soluble and membrane proteins

    International Nuclear Information System (INIS)

    Huang, Chia-Ying; Olieric, Vincent; Ma, Pikyee; Panepucci, Ezequiel; Diederichs, Kay; Wang, Meitian; Caffrey, Martin

    2015-01-01

    A method for performing high-throughput in situ serial X-ray crystallography with soluble and membrane proteins in the lipid cubic phase is described. It works with microgram quantities of protein and lipid (and ligand when present) and is compatible with the most demanding sulfur SAD phasing. The lipid cubic phase (LCP) continues to grow in popularity as a medium in which to generate crystals of membrane (and soluble) proteins for high-resolution X-ray crystallographic structure determination. To date, the PDB includes 227 records attributed to the LCP or in meso method. Among the listings are some of the highest profile membrane proteins, including the β 2 -adrenoreceptor–G s protein complex that figured in the award of the 2012 Nobel Prize in Chemistry to Lefkowitz and Kobilka. The most successful in meso protocol to date uses glass sandwich crystallization plates. Despite their many advantages, glass plates are challenging to harvest crystals from. However, performing in situ X-ray diffraction measurements with these plates is not practical. Here, an alternative approach is described that provides many of the advantages of glass plates and is compatible with high-throughput in situ measurements. The novel in meso in situ serial crystallography (IMISX) method introduced here has been demonstrated with AlgE and PepT (alginate and peptide transporters, respectively) as model integral membrane proteins and with lysozyme as a test soluble protein. Structures were solved by molecular replacement and by experimental phasing using bromine SAD and native sulfur SAD methods to resolutions ranging from 1.8 to 2.8 Å using single-digit microgram quantities of protein. That sulfur SAD phasing worked is testament to the exceptional quality of the IMISX diffraction data. The IMISX method is compatible with readily available, inexpensive materials and equipment, is simple to implement and is compatible with high-throughput in situ serial data collection at macromolecular

  19. THE PRODUCTION RATE OF SN Ia EVENTS IN GLOBULAR CLUSTERS

    International Nuclear Information System (INIS)

    Washabaugh, Pearce C.; Bregman, Joel N.

    2013-01-01

    In globular clusters, dynamical evolution produces luminous X-ray emitting binaries at a rate about 200 times greater than in the field. If globular clusters also produce SN Ia at a high rate, it would account for many of the SN Ia production in early-type galaxies and provide insight into their formation. Here we use archival Hubble Space Telescope (HST) images of nearby galaxies that have hosted an SN Ia to examine the rate at which globular clusters produce these events. The location of the SN Ia is registered on an HST image obtained before the event or after the supernova (SN) faded. Of the 36 nearby galaxies examined, 21 had sufficiently good data to search for globular cluster hosts. None of the 21 SNe have a definite globular cluster counterpart, although there are some ambiguous cases. This places an upper limit to the enhancement rate of SN Ia production in globular clusters of about 42 at the 95% confidence level, which is an order of magnitude lower than the enhancement rate for luminous X-ray binaries. Even if all of the ambiguous cases are considered as having a globular cluster counterpart, the upper bound for the enhancement rate is 82 at the 95% confidence level, still a factor of several below that needed to account for half of the SN Ia events. Barring unforeseen selection effects, we conclude that globular clusters are not responsible for producing a significant fraction of the SN Ia events in early-type galaxies.

  20. Effects of Toasting Time on Digestive Hydrolysis of Soluble and Insoluble 00-Rapeseed Meal Proteins

    NARCIS (Netherlands)

    Salazar-Villanea, Sergio; Bruininx, Erik M.A.M.; Gruppen, Harry; Carré, Patrick; Quinsac, Alain; Poel, van der Thomas

    2017-01-01

    Thermal damage to proteins can reduce their nutritional value. The effects of toasting time on the kinetics of hydrolysis, the resulting molecular weight distribution of 00-rapeseed meal (RSM) and the soluble and insoluble protein fractions separated from the RSM were studied. Hydrolysis was

  1. Rapid Determination of Protein Solubility and Stability Conditions for NMR Studies Using Incomplete Factorial Design

    International Nuclear Information System (INIS)

    Ducat, Thierry; Declerck, Nathalie; Gostan, Thierry; Kochoyan, Michel; Demene, Helene

    2006-01-01

    Sample preparation constitutes a crucial and limiting step in structural studies of proteins by NMR. The determination of the solubility and stability (SAS) conditions of biomolecules at millimolar concentrations stays today empirical and hence time- and material-consuming. Only few studies have been recently done in this field and they have highlighted the interest of using crystallogenesis tools to optimise sample conditions. In this study, we have adapted a method based on incomplete factorial design and making use of crystallisation plates to quantify the influence of physico-chemical parameters such as buffer pH and salts on protein SAS. A description of the experimental set up and an evaluation of the method are given by case studies on two functional domains from the bacterial regulatory protein LicT as well as two other proteins. Using this method, we could rapidly determine optimised conditions for extracting soluble proteins from bacterial cells and for preparing purified protein samples sufficiently concentrated and stable for NMR characterisation. The drastic reduction in the time and number of experiments required for searching protein SAS conditions makes this method particularly well-adapted for a systematic investigation on a large range of physico-chemical parameters

  2. Protein scissors: Photocleavage of proteins at specific locations

    Indian Academy of Sciences (India)

    Unknown

    Binding of ligands to globular proteins at hydrophobic cavities while making specific ... ched to a PTI model A1010 monochromator. UV cut-off filter ..... >1:1 stoichiometry (protein to ligand), the binding equilibrium favors the thermo- dynamically ...

  3. Structural proteomics of minimal organisms: conservation ofprotein fold usage and evolutionary implications

    Energy Technology Data Exchange (ETDEWEB)

    Chandonia, John-Marc; Kim, Sung-Hou

    2006-03-15

    Background: Determining the complete repertoire of proteinstructures for all soluble, globular proteins in a single organism hasbeen one of the major goals of several structural genomics projects inrecent years. Results: We report that this goal has nearly been reachedfor several "minimal organisms"--parasites or symbionts with reducedgenomes--for which over 95 percent of the soluble, globular proteins maynow be assigned folds, overall 3-D backbone structures. We analyze thestructures of these proteins as they relate to cellular functions, andcompare conservation off old usage between functional categories. We alsocompare patterns in the conservation off olds among minimal organisms andthose observed between minimal organisms and other bacteria. Conclusion:We find that proteins performing essential cellular functions closelyrelated to transcription and translation exhibit a higher degree ofconservation in fold usage than proteins in other functional categories.Folds related to transcription and translation functional categories werealso over represented in minimal organisms compared to otherbacteria.

  4. Soluble expression of recomb inant cMyc, Klf4, Oct4, and Sox2 proteins in bacteria and transduction into living cells

    Directory of Open Access Journals (Sweden)

    Guo-Dan Liu

    2017-04-01

    Full Text Available AIM: To develop a new method to produce recombinant reprogramming proteins, cMyc, Klf4, Oct4, and Sox2, in soluble format with low cost for the generation of induced pluripotent stem cells (iPSCs. METHODS: A short polypeptide sequence derived from the HIV trans-activator of transcription protein (TAT and the nucleus localization signal (NLS polypeptide were fused to the N terminus of the reprogramming proteins and they were constructed into pCold-SUMO vector which can extremely improve the solubility of recombinant proteins. Then these vector plasmids were transformed into E. coli BL21 (DE3 Chaperone competent cells for amplification. The solubility of these recombinant proteins was determined by SDS-PAGE and Coomassie brilliant blue staining. The recombinant proteins were purified by Ni-NTA resin and identified by Western blot. The transduction of these proteins into HEK 293T cells were evaluated by immunofluorescence staining. RESULTS: These four reprogramming proteins could be produced in soluble format in pCold-SUMO expression vector system with the assistance of chaperone proteins in bacteria. The proteins were purified successfully with a purity of over 70% with a relative high transduction rate into 293 cells. CONCLUSION: The results in the present study indicate the four important reprogramming proteins, cMyc, Klf4, Oct4, and Sox2, can be produced in soluble format in bacteria with low cost. Our new method thus might be expected to greatly contribute to the future study of iPSCs.

  5. Access to gram scale amounts of functional globular adiponectin from E. coli inclusion bodies by alkaline-shock solubilization.

    Science.gov (United States)

    Heiker, John T; Klöting, Nora; Blüher, Matthias; Beck-Sickinger, Annette G

    2010-07-16

    The adipose tissue derived protein adiponectin exerts anti-diabetic, anti-inflammatory and anti-atherosclerotic effects. Adiponectin serum concentrations are in the microgram per milliliter range in healthy humans and inversely correlate with obesity and metabolic disorders. Accordingly, raising circulating adiponectin levels by direct administration may be an intriguing strategy in the treatment of obesity-related metabolic disorders. However production of large amounts of recombinant adiponectin protein is a primary obstacle so far. Here, we report a novel method for large amount production of globular adiponectin from E. coli inclusion bodies utilizing an alkaline-shock solubilization method without chaotropic agents followed by precipitation of the readily renaturing protein. Precipitation of the mildly solubilized protein capitalizes on advantages of inclusion body formation. This approach of inclusion body protein recovery provides access to gram scale amounts of globular adiponectin with standard laboratory equipment avoiding vast dilution or dialysis steps to neutralize the pH and renature the protein, thus saving chemicals and time. The precipitated protein is readily renaturing in buffer, is of adequate purity without a chromatography step and shows biological activity in cultured MCF7 cells and significantly lowered blood glucose levels in mice with streptozotocin induced type 1 diabetes. Copyright 2010 Elsevier Inc. All rights reserved.

  6. TOWARDS THE CONCEPT OF GLOBULAR PEARLITE

    Directory of Open Access Journals (Sweden)

    A. G. Anisovich

    2016-01-01

    Full Text Available Explanatory imprecisions of concept of globular pearlite and ferrite-carbide-mixture are considered. The need of concept binding of globular pearlite to specific grain with 0.8% carbon content is explained with the assistance of exemplary data obtained at the present metallographic equipment. The question of educational material presentation concerning the process of teaching of discipline «Materials and construction materials technology» is discussed in relation to the educational process of technical universities, in particular, the Belarusian State Agrarian Technical University.

  7. Implications of protein polymorphism on protein phase behaviour

    NARCIS (Netherlands)

    Stegen, J.; Schoot, van der P.P.A.M.

    2015-01-01

    The phase behaviour of small globular proteins is often modeled by approximating them as spherical particles with fixed internal structure. However, changes in the local environment of a protein can lead to changes in its conformation rendering this approximation invalid. We present a simple

  8. Effect of plaster cast immobilization on the turnover rates of soluble proteins and lactate dehydrogenase isoenzymes of rabbit M. soleus

    Energy Technology Data Exchange (ETDEWEB)

    Edes, I.; Dosa, E.; Sohar, I.; Guba, F. (Orvostudomanyi Egyetem, Szeged (Hungary). Biokemiai Tanszek)

    1982-01-01

    In atrophized muscle the decreases of the activity of LDH isoenzymes can be explained partly by a 15 per cent decrease of the enzyme synthesis and partly by a 25 per cent increase in catabolism. The quantities of the soluble proteins and LDH were measured after intravenously administered /sup 3/H-leucin incorporation, from the musculus soleus. LDH was isolated by means of affinity chromatography. Radioactivity was determined in a Packard Tri-Carb scintillation counter. The synthesis rate of soluble proteins barely changed during immobilization. In the atrophized muscle the decrease of the amount of soluble proteins could be almost exclusively interpreted in terms of a 25 per cent enchancement of degradative process. The accelerated catabolism is most probably due to the proteolytic enzymes activated by immobilization.

  9. Pulsar-irradiated stars in dense globular clusters

    Science.gov (United States)

    Tavani, Marco

    1992-01-01

    We discuss the properties of stars irradiated by millisecond pulsars in 'hard' binaries of dense globular clusters. Irradiation by a relativistic pulsar wind as in the case of the eclipsing millisecond pulsar PSR 1957+20 alter both the magnitude and color of the companion star. Some of the blue stragglers (BSs) recently discovered in dense globular clusters can be irradiated stars in binaries containing powerful millisecond pulsars. The discovery of pulsar-driven orbital modulations of BS brightness and color with periods of a few hours together with evidence for radio and/or gamma-ray emission from BS binaries would valuably contribute to the understanding of the evolution of collapsed stars in globular clusters. Pulsar-driven optical modulation of cluster stars might be the only observable effect of a new class of binary pulsars, i.e., hidden millisecond pulsars enshrouded in the evaporated material lifted off from the irradiated companion star.

  10. Possible systematic decreases in the age of globular clusters

    Energy Technology Data Exchange (ETDEWEB)

    Shi, X. [Univ. of Chicago, Chicago, IL (United States); Schramm, D. N. [Fermi National Accelerator Laboratory (FNAL), Batavia, IL (United States); Univ. of Chicago, Chicago, IL (United States); Dearborn, D. S.P. [Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States); Truran, J. W. [Univ. of Chicago, Chicago, IL (United States)

    1994-03-01

    The ages of globular clusters inferred from observations depends sensitively on assumptions like the initial helium abundance and the mass loss rate. A high helium abundance (e.g., Y\\approx0.28) or a mass loss rate of \\sim10^{-11}M_\\odot yr^{-1} near the main sequence turn-off region lowers the current age estimate from 14 Gyr to about 10--12 Gyr, significantly relaxing the constraints on the Hubble constant, allowing values as high as 60km/sec/Mpc for a universe with the critical density and 90km/sec/Mpc for a baryon-only universe. Possible mechanisms for the helium enhancement in globular clusters are discussed, as are arguments for an instability strip induced mass loss near the turn-off. Ages lower than 10 Gyr are not possible even with the operation of both of these mechanisms unless the initial helium abundance in globular clusters is >0.30, which would conflict with indirect measurements of helium abundances in globular clusters.

  11. Cleaved thioredoxin fusion protein enables the crystallization of poorly soluble ERα in complex with synthetic ligands

    International Nuclear Information System (INIS)

    Cura, Vincent; Gangloff, Monique; Eiler, Sylvia; Moras, Dino; Ruff, Marc

    2007-01-01

    A new crystallization strategy: the presence of cleaved thioredoxin fusion is critical for crystallization of the estrogen nuclear receptor ligand binding domain in complex with synthetic ligands. This novel technique should be regarded as an interesting alternative for crystallization of difficult proteins. The ligand-binding domain (LBD) of human oestrogen receptor α was produced in Escherichia coli as a cleavable thioredoxin (Trx) fusion in order to improve solubility. Crystallization trials with either cleaved and purified LBD or with the purified fusion protein both failed to produce crystals. In another attempt, Trx was not removed from the LBD after endoproteolytic cleavage and its presence promoted nucleation and subsequent crystal growth, which allowed the structure determination of two different LBD–ligand–coactivator peptide complexes at 2.3 Å resolution. This technique is likely to be applicable to other low-solubility proteins

  12. Effect of the horizontal branch on the colours of globular clusters

    Energy Technology Data Exchange (ETDEWEB)

    Sil' chenko, O K [Moskovskij Gosudarstvennyj Univ. (USSR). Gosudarstvennyj Astronomicheskij Inst. ' ' GAISh' '

    1963-05-01

    The influence of the horizontal branch (HB) on the integral UBV colours of globular clusters is studied by means of statistical analysis of the colour-magnitude diagram catalogue for globular clusters of our Galaxy. The colour correction for HB is shown to be always negative. It turns out to be small for m. tal-rich globular clusters ((Fe/H)>-1.1) and independent on the HB shape for metal-poor ones.

  13. Effect of the horizontal branch on the colours of globular clusters

    International Nuclear Information System (INIS)

    Sil'chenko, O.K.

    1963-01-01

    The influence of the horizontal branch (HB) on the integral UBV colours of globular clusters is studied by means of statistical analysis of the colour-magnitude diagram catalogue for globular clusters of our Galaxy. The colour correction for HB is shown to be always negative. It turns out to be small for m. tal-rich globular clusters ([Fe/H]>-1.1) and independent on the HB shape for metal-poor ones

  14. The WAGGS project - I. The WiFeS Atlas of Galactic Globular cluster Spectra

    Science.gov (United States)

    Usher, Christopher; Pastorello, Nicola; Bellstedt, Sabine; Alabi, Adebusola; Cerulo, Pierluigi; Chevalier, Leonie; Fraser-McKelvie, Amelia; Penny, Samantha; Foster, Caroline; McDermid, Richard M.; Schiavon, Ricardo P.; Villaume, Alexa

    2017-07-01

    We present the WiFeS Atlas of Galactic Globular cluster Spectra, a library of integrated spectra of Milky Way and Local Group globular clusters. We used the WiFeS integral field spectrograph on the Australian National University 2.3 m telescope to observe the central regions of 64 Milky Way globular clusters and 22 globular clusters hosted by the Milky Way's low-mass satellite galaxies. The spectra have wider wavelength coverage (3300-9050 Å) and higher spectral resolution (R = 6800) than existing spectral libraries of Milky Way globular clusters. By including Large and Small Magellanic Cloud star clusters, we extend the coverage of parameter space of existing libraries towards young and intermediate ages. While testing stellar population synthesis models and analysis techniques is the main aim of this library, the observations may also further our understanding of the stellar populations of Local Group globular clusters and make possible the direct comparison of extragalactic globular cluster integrated light observations with well-understood globular clusters in the Milky Way. The integrated spectra are publicly available via the project website.

  15. Synthesis of acid-soluble spore proteins by Bacillus subtilis.

    Science.gov (United States)

    Leventhal, J M; Chambliss, G H

    1982-12-01

    The major acid-soluble spore proteins (ASSPs) of Bacillus subtilis were detected by immunoprecipitation of radioactively labeled in vitro- and in vivo-synthesized proteins. ASSP synthesis in vivo began 2 h after the initiation of sporulation (t2) and reached its maximum rate at t7. This corresponded to the time of synthesis of mRNA that stimulated the maximum rate of ASSP synthesis in vitro. Under the set of conditions used in these experiments, protease synthesis began near t0, alkaline phosphatase synthesis began at about t2, and refractile spores were first observed between t7 and t8. In vivo- and in vitro-synthesized ASSPs comigrated in sodium dodecyl sulfate-polyacrylamide gels. Their molecular weights were 4,600 (alpha and beta) and 11,000 (gamma). The average half-life of the ASSP messages was 11 min when either rifampin (10 micrograms/ml) or actinomycin D (1 microgram/ml) was used to inhibit RNA synthesis.

  16. An electrophoretic study of the soluble lens proteins from the Indian mackerel, Rastrelliger kanagurta (Cuv)

    Digital Repository Service at National Institute of Oceanography (India)

    Menezes, M.R.

    Soluble eye lens nuclei proteins of the Indian mackerel Rastrelliger kanagurta were studied by cellogel electrophoresis, to see whether there are any intra species variations. A distinct pattern characterised by the number of bands, mobility...

  17. Dynamic prestress in a globular protein.

    Directory of Open Access Journals (Sweden)

    Scott A Edwards

    Full Text Available A protein at equilibrium is commonly thought of as a fully relaxed structure, with the intra-molecular interactions showing fluctuations around their energy minimum. In contrast, here we find direct evidence for a protein as a molecular tensegrity structure, comprising a balance of tensed and compressed interactions, a concept that has been put forward for macroscopic structures. We quantified the distribution of inter-residue prestress in ubiquitin and immunoglobulin from all-atom molecular dynamics simulations. The network of highly fluctuating yet significant inter-residue forces in proteins is a consequence of the intrinsic frustration of a protein when sampling its rugged energy landscape. In beta sheets, this balance of forces is found to compress the intra-strand hydrogen bonds. We estimate that the observed magnitude of this pre-compression is enough to induce significant changes in the hydrogen bond lifetimes; thus, prestress, which can be as high as a few 100 pN, can be considered a key factor in determining the unfolding kinetics and pathway of proteins under force. Strong pre-tension in certain salt bridges on the other hand is connected to the thermodynamic stability of ubiquitin. Effective force profiles between some side-chains reveal the signature of multiple, distinct conformational states, and such static disorder could be one factor explaining the growing body of experiments revealing non-exponential unfolding kinetics of proteins. The design of prestress distributions in engineering proteins promises to be a new tool for tailoring the mechanical properties of made-to-order nanomaterials.

  18. Protein shedding in urothelial bladder cancer: prognostic implications of soluble urinary EGFR and EpCAM.

    Science.gov (United States)

    Bryan, R T; Regan, H L; Pirrie, S J; Devall, A J; Cheng, K K; Zeegers, M P; James, N D; Knowles, M A; Ward, D G

    2015-03-17

    Better biomarkers must be found to develop clinically useful urine tests for bladder cancer. Proteomics can be used to identify the proteins released by cancer cell lines and generate candidate markers for developing such tests. We used shotgun proteomics to identify proteins released into culture media by eight bladder cancer cell lines. These data were compared with protein expression data from the Human Protein Atlas. Epidermal growth factor receptor (EGFR) was identified as a candidate biomarker and measured by ELISA in urine from 60 noncancer control subjects and from 436 patients with bladder cancer and long-term clinical follow-up. Bladder cancer cell lines shed soluble EGFR ectodomain. Soluble EGFR is also detectable in urine and is highly elevated in some patients with high-grade bladder cancer. Urinary EGFR is an independent indicator of poor bladder cancer-specific survival with a hazard ratio of 2.89 (95% CI 1.81-4.62, Pbladder cancer-specific survival and have prognostic value over and above that provided by standard clinical observations. Measuring urinary EGFR and EpCAM may represent a simple and useful approach for fast-tracking the investigation and treatment of patients with the most aggressive bladder cancers.

  19. Millisecond radio pulsars in globular clusters

    Science.gov (United States)

    Verbunt, Frank; Lewin, Walter H. G.; Van Paradijs, Jan

    1989-01-01

    It is shown that the number of millisecond radio pulsars, in globular clusters, should be larger than 100, applying the standard scenario that all the pulsars descend from low-mass X-ray binaries. Moreover, most of the pulsars are located in a small number of clusters. The prediction that Teran 5 and Liller 1 contain at least about a dozen millisecond radio pulsars each is made. The observations of millisecond radio pulsars in globular clusters to date, in particular the discovery of two millisecond radio pulsars in 47 Tuc, are in agreement with the standard scenario, in which the neutron star is spun up during the mass transfer phase.

  20. Globular cluster metallicity scale: evidence from stellar models

    International Nuclear Information System (INIS)

    Demarque, P.; King, C.R.; Diaz, A.

    1982-01-01

    Theoretical giant branches have been constructed to determine their relative positions for metallicities in the range -2.3 0 )/sub 0,g/ based on these models is presented which yields good agreement over the observed range of metallicities for galactic globular clusters and old disk clusters. The metallicity of 47 Tuc and M71 given by this calibration is about -0.8 dex. Subject headings: clusters, globular: stars: abundances: stars: interiors

  1. Heat-Induced Soluble Protein Aggregates from Mixed Pea Globulins and β-Lactoglobulin.

    Science.gov (United States)

    Chihi, Mohamed-Lazhar; Mession, Jean-luc; Sok, Nicolas; Saurel, Rémi

    2016-04-06

    The present work investigates the formation of protein aggregates (85 °C, 60 min incubation) upon heat treatment of β-lactoglobulin (βlg)-pea globulins (Glob) mixtures at pH 7.2 and 5 mM NaCl from laboratory-prepared protein isolates. Various βlg/Glob weight ratios were applied, for a total protein concentration of 2 wt % in admixture. Different analytical methods were used to determine the aggregation behavior of "mixed" aggregates, that is, surface hydrophobicity and also sulfhydryl content, protein interactions by means of SDS-PAGE electrophoresis, and molecule size distribution by DLS and gel filtration. The production of "mixed" thermal aggregates would involve both the formation of new disulfide bonds and noncovalent interactions between the denatured βlg and Glob subunits. The majority of "mixed" soluble aggregates displayed higher molecular weight and smaller diameter than those for Glob heated in isolation. The development of pea-whey protein "mixed" aggregates may help to design new ingredients for the control of innovative food textures.

  2. Water-Soluble Dried Blood Spot in Protein Analysis: A Proof-of-Concept Study.

    Science.gov (United States)

    Rosting, Cecilie; Gjelstad, Astrid; Halvorsen, Trine Grønhaug

    2015-08-04

    In the present work human chorionic gonadotropin (hCG) was used as a model protein in a proof-of-concept study combining water-soluble dried blood spot (DBS) material in liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based protein analysis. A water-soluble material consisting of commercially available carboxymethyl cellulose (CMC) was evaluated as sampling material for this purpose. The material dissolved readily at physiological pH. Different sample preparation methods were evaluated, and in the final method, 15 μL of whole blood was deposited and dried on CMC before the whole spot was dissolved prior to cleanup by immunoaffinity extraction, tryptic digest, and preconcentration by solid-phase extraction (SPE). The results indicated complete dissolution of hCG from the spots, acceptable limit of detection (LOD) (0.1 IU/mL), linearity (R(2) = 0.959), accuracy (16%), and precision (≤22%). Long-term stability (45 days) of hCG in dried spots at reduced temperatures (≤8 °C) was also demonstrated. The analyte recovery was comparable to the commercially available nonsolvable cellulose material (FTA DMPK-C card).

  3. Soluble arabinoxylan alters digesta flow and protein digestion of red meat-containing diets in pigs.

    Science.gov (United States)

    Zhang, Dagong; Williams, Barbara A; Mikkelsen, Deirdre; Li, Xiuhua; Keates, Helen L; Lisle, Allan T; Collins, Helen M; Fincher, Geoffrey B; Bird, Anthony R; Topping, David L; Gidley, Michael J; Bryden, Wayne L

    2015-09-01

    The aim of this study was to investigate how a moderate increase in dietary meat content combined (or not) with soluble fibre would influence protein digestion as well as digesta characteristics and flow. Four groups of pigs were fed Western-style diets (high-protein/high-fat) containing two types of barbecued red meat, one with and one without a wheat arabinoxylan-rich fraction. After 4 wk, digesta samples were collected from small and large intestinal sites and analyzed for protein, amino acids, dry matter, and acid-insoluble ash. Tissue samples were also collected from each site. Arabinoxylan consumption led to somewhat lower apparent protein digestibility within the small and large intestines as well as shorter mean retention times. This suggests that the lowered protein digestibility is due, at least partly, to shorter access time to digestive proteases and absorptive surfaces. Additionally, digesta mass was higher in pigs fed arabinoxylan while dry matter (%) was lower, indicating an increased digesta water-holding capacity due to the presence of a soluble dietary fiber. Data showed that solubilized wheat arabinoxylan provides potential health benefits through decreased protein digestibility, increased digesta mass, and reduced mean retention time, even for diets with a moderately higher protein content. These factors are associated with efficiency of digestion and satiety, both of which have implications for prevention of obesity and other health disorders. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Supra-galactic colour patterns in globular cluster systems

    Science.gov (United States)

    Forte, Juan C.

    2017-07-01

    An analysis of globular cluster systems associated with galaxies included in the Virgo and Fornax Hubble Space Telescope-Advanced Camera Surveys reveals distinct (g - z) colour modulation patterns. These features appear on composite samples of globular clusters and, most evidently, in galaxies with absolute magnitudes Mg in the range from -20.2 to -19.2. These colour modulations are also detectable on some samples of globular clusters in the central galaxies NGC 1399 and NGC 4486 (and confirmed on data sets obtained with different instruments and photometric systems), as well as in other bright galaxies in these clusters. After discarding field contamination, photometric errors and statistical effects, we conclude that these supra-galactic colour patterns are real and reflect some previously unknown characteristic. These features suggest that the globular cluster formation process was not entirely stochastic but included a fraction of clusters that formed in a rather synchronized fashion over large spatial scales, and in a tentative time lapse of about 1.5 Gy at redshifts z between 2 and 4. We speculate that the putative mechanism leading to that synchronism may be associated with large scale feedback effects connected with violent star-forming events and/or with supermassive black holes.

  5. Images in the rocket ultraviolet - UV fluxes of M31 globular clusters

    International Nuclear Information System (INIS)

    Bohlin, R.C.; Cornett, R.H.; Hill, J.K.; Hill, R.S.; Stecher, T.P.

    1988-01-01

    Images obtained by a rocket-borne UV imaging telescope are used here to determine near-UV fluxes for 17 sources in M31 that are optical globular-cluster candidates and for the bright open cluster vdB0 in M31. Far-UV fluxes or flux limits are determined for the same clusters. The m(NUV)-V colors for M31 clusters are similar to those of Galactic clusters, except for the high-metallicity M31 cluster Bo 171. Four of the detected clusters have optical, m(NUV) - V, and m(FUV) - V colors indicating ages of about 100 million years. These four clusters are probably similar to the so-called 'blue globular' clusters of the LMC. The existence of young LMC-type blue globulars and the possible existence of middle-aged metal-rich globulars may indicate that M31 has continued to form globular clusters throughout its life. 39 references

  6. Utilization of paramagnetic relaxation enhancements for high-resolution NMR structure determination of a soluble loop-rich protein with sparse NOE distance restraints

    International Nuclear Information System (INIS)

    Furuita, Kyoko; Kataoka, Saori; Sugiki, Toshihiko; Hattori, Yoshikazu; Kobayashi, Naohiro; Ikegami, Takahisa; Shiozaki, Kazuhiro; Fujiwara, Toshimichi; Kojima, Chojiro

    2015-01-01

    NMR structure determination of soluble proteins depends in large part on distance restraints derived from NOE. In this study, we examined the impact of paramagnetic relaxation enhancement (PRE)-derived distance restraints on protein structure determination. A high-resolution structure of the loop-rich soluble protein Sin1 could not be determined by conventional NOE-based procedures due to an insufficient number of NOE restraints. By using the 867 PRE-derived distance restraints obtained from the NOE-based structure determination procedure, a high-resolution structure of Sin1 could be successfully determined. The convergence and accuracy of the determined structure were improved by increasing the number of PRE-derived distance restraints. This study demonstrates that PRE-derived distance restraints are useful in the determination of a high-resolution structure of a soluble protein when the number of NOE constraints is insufficient

  7. Soluble proteins from fowl feather keratin. II. Isolation of some proteins from barbs.

    Science.gov (United States)

    Murayama, K; Akahane, K; Murozono, S

    1977-01-01

    Four fractions, GF-1, 2, 3, and 4, which had been separated from S-carboxymethylated (SCM-) proteins of fowl feathers by gel filtration, were each chromatographed on a DEAE-cellulose column in 0.05 M Tris-HCl buffer (pH 8.5) containing 8 M urea. The major fraction, GF-3, was further separated into seven peaks; the first four were shown to be single components by polyacrylamide disc gel electrophoresis. Chromatograms of GF-1 and 2 showed broad peaks which appeared at nearly the same volume as in GF-3. The components from GF-3 had very similar amino acid compositions except that the SCM-cysteine content showed a tendency to increase in the order of elution from the column. SCM-extract prepared from barbs of the wing feathers of a fowl was more heterogeneous than that taken from the body feathers. A combination of gel filtration on Sephadex G-75 and chromatography on DEAE-cellulose was found to be more effective for the isolation of soluble SCM-proteins.

  8. Individual globular domains and domain unfolding visualized in overstretched titin molecules with atomic force microscopy.

    Directory of Open Access Journals (Sweden)

    Zsolt Mártonfalvi

    Full Text Available Titin is a giant elastomeric protein responsible for the generation of passive muscle force. Mechanical force unfolds titin's globular domains, but the exact structure of the overstretched titin molecule is not known. Here we analyzed, by using high-resolution atomic force microscopy, the structure of titin molecules overstretched with receding meniscus. The axial contour of the molecules was interrupted by topographical gaps with a mean width of 27.7 nm that corresponds well to the length of an unfolded globular (immunoglobulin and fibronectin domain. The wide gap-width distribution suggests, however, that additional mechanisms such as partial domain unfolding and the unfolding of neighboring domain multimers may also be present. In the folded regions we resolved globules with an average spacing of 5.9 nm, which is consistent with a titin chain composed globular domains with extended interdomain linker regions. Topographical analysis allowed us to allocate the most distal unfolded titin region to the kinase domain, suggesting that this domain systematically unfolds when the molecule is exposed to overstretching forces. The observations support the prediction that upon the action of stretching forces the N-terminal ß-sheet of the titin kinase unfolds, thus exposing the enzyme's ATP-binding site and hence contributing to the molecule's mechanosensory function.

  9. A novel water-based process produces eco-friendly bio-adhesive made from green cross-linked soybean soluble polysaccharide and soy protein.

    Science.gov (United States)

    Yuan, Cheng; Chen, Mingsong; Luo, Jing; Li, Xiaona; Gao, Qiang; Li, Jianzhang

    2017-08-01

    In this study, an eco-friendly soy protein adhesive was developed that utilized two components from soybean meal without addition of any toxic material. A plant-based, water-soluble and inexpensive soybean soluble polysaccharide was used as the novel renewable material to combine with soy protein to produce a soy protein adhesive. Three-plywood was fabricated with the resulting adhesive, and its wet shear strength was measured. The results showed the wet shear strength of plywood bonded by the adhesive reached 0.99MPa, meeting the water resistance requirement for interior use panels. This improvement was attributed to the following reasons: (1) Combination of cross-linked soybean soluble polysaccharide and soy protein formed an interpenetrating network structure, improving the thermal stability and water resistance of the cured adhesive. (2) Adding CL-SSPS decreased the adhesive viscosity to 15.14Pas, which increased the amount of the adhesive that penetrate the wood's surface and formed more interlocks. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Exploring the Internal Dynamics of Globular Clusters

    Science.gov (United States)

    Watkins, Laura L.; van der Marel, Roeland; Bellini, Andrea; Luetzgendorf, Nora; HSTPROMO Collaboration

    2018-01-01

    Exploring the Internal Dynamics of Globular ClustersThe formation histories and structural properties of globular clusters are imprinted on their internal dynamics. Energy equipartition results in velocity differences for stars of different mass, and leads to mass segregation, which results in different spatial distributions for stars of different mass. Intermediate-mass black holes significantly increase the velocity dispersions at the centres of clusters. By combining accurate measurements of their internal kinematics with state-of-the-art dynamical models, we can characterise both the velocity dispersion and mass profiles of clusters, tease apart the different effects, and understand how clusters may have formed and evolved.Using proper motions from the Hubble Space Telescope Proper Motion (HSTPROMO) Collaboration for a set of 22 Milky Way globular clusters, and our discrete dynamical modelling techniques designed to work with large, high-quality datasets, we are studying a variety of internal cluster properties. We will present the results of theoretical work on simulated clusters that demonstrates the efficacy of our approach, and preliminary results from application to real clusters.

  11. Properties of whey protein isolates extruded under acidic and alkaline conditions.

    Science.gov (United States)

    Onwulata, C I; Isobe, S; Tomasula, P M; Cooke, P H

    2006-01-01

    Whey proteins have wide acceptance and use in many products due to their beneficial nutritional properties. To further increase the amount of whey protein isolates (WPI) that may be added to products such as extruded snacks and meats, texturization of WPI is necessary. Texturization changes the folding of globular proteins to improve interaction with other ingredients and create new functional ingredients. In this study, WPI pastes (60% solids) were extruded in a twin-screw extruder at 100 degrees C with 4 pH-adjusted water streams: acidic (pH 2.0 +/- 0.2) and alkaline (pH 12.4 +/- 0.4) streams from 2 N HCl and 2 N NaOH, respectively, and acidic (pH 2.5 +/- 0.2) and alkaline (pH 11.5 +/- 0.4) electrolyzed water streams; these were compared with WPI extruded with deionized water. The effects of water acidity on WPI solubility at pH 7, color, microstructure, Rapid Visco Analyzer pasting properties, and physical structure were determined. Alkaline conditions increased insolubility caused yellowing and increased pasting properties significantly. Acidic conditions increased solubility and decreased WPI pasting properties. Subtle structural changes occurred under acidic conditions, but were more pronounced under alkaline conditions. Overall, alkaline conditions increased denaturation in the extruded WPI resulting in stringy texturized WPI products, which could be used in meat applications.

  12. DARK MATTER HALOS IN GALAXIES AND GLOBULAR CLUSTER POPULATIONS

    Energy Technology Data Exchange (ETDEWEB)

    Hudson, Michael J.; Harris, Gretchen L. [Department of Physics and Astronomy, University of Waterloo, Waterloo, ON N2L 3G1 (Canada); Harris, William E., E-mail: mjhudson@uwaterloo.ca [Department of Physics and Astronomy, McMaster University, Hamilton, ON L8S 4M1 (Canada)

    2014-05-20

    We combine a new, comprehensive database for globular cluster populations in all types of galaxies with a new calibration of galaxy halo masses based entirely on weak lensing. Correlating these two sets of data, we find that the mass ratio η ≡ M {sub GCS}/M {sub h} (total mass in globular clusters, divided by halo mass) is essentially constant at (η) ∼ 4 × 10{sup –5}, strongly confirming earlier suggestions in the literature. Globular clusters are the only known stellar population that formed in essentially direct proportion to host galaxy halo mass. The intrinsic scatter in η appears to be at most 0.2 dex; we argue that some of this scatter is due to differing degrees of tidal stripping of the globular cluster systems between central and satellite galaxies. We suggest that this correlation can be understood if most globular clusters form at very early stages in galaxy evolution, largely avoiding the feedback processes that inhibited the bulk of field-star formation in their host galaxies. The actual mean value of η also suggests that about one-fourth of the initial gas mass present in protogalaxies collected into giant molecular clouds large enough to form massive, dense star clusters. Finally, our calibration of (η) indicates that the halo masses of the Milky Way and M31 are (1.2 ± 0.5) × 10{sup 12} M {sub ☉} and (3.9 ± 1.8) × 10{sup 12} M {sub ☉}, respectively.

  13. RETENTION OF STELLAR-MASS BLACK HOLES IN GLOBULAR CLUSTERS

    International Nuclear Information System (INIS)

    Morscher, Meagan; Umbreit, Stefan; Farr, Will M.; Rasio, Frederic A.

    2013-01-01

    Globular clusters should be born with significant numbers of stellar-mass black holes (BHs). It has been thought for two decades that very few of these BHs could be retained through the cluster lifetime. With masses ∼10 M ☉ , BHs are ∼20 times more massive than an average cluster star. They segregate into the cluster core, where they may eventually decouple from the remainder of the cluster. The small-N core then evaporates on a short timescale. This is the so-called Spitzer instability. Here we present the results of a full dynamical simulation of a globular cluster containing many stellar-mass BHs with a realistic mass spectrum. Our Monte Carlo simulation code includes detailed treatments of all relevant stellar evolution and dynamical processes. Our main finding is that old globular clusters could still contain many BHs at present. In our simulation, we find no evidence for the Spitzer instability. Instead, most of the BHs remain well mixed with the rest of the cluster, with only the innermost few tens of BHs segregating significantly. Over the 12 Gyr evolution, fewer than half of the BHs are dynamically ejected through strong binary interactions in the cluster core. The presence of BHs leads to long-term heating of the cluster, ultimately producing a core radius on the high end of the distribution for Milky Way globular clusters (and those of other galaxies). A crude extrapolation from our model suggests that the BH-BH merger rate from globular clusters could be comparable to the rate in the field.

  14. Surface properties of heat-induced soluble soy protein aggregates of different molecular masses.

    Science.gov (United States)

    Guo, Fengxian; Xiong, Youling L; Qin, Fang; Jian, Huajun; Huang, Xiaolin; Chen, Jie

    2015-02-01

    Suspensions (2% and 5%, w/v) of soy protein isolate (SPI) were heated at 80, 90, or 100 °C for different time periods to produce soluble aggregates of different molecular sizes to investigate the relationship between particle size and surface properties (emulsions and foams). Soluble aggregates generated in these model systems were characterized by gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Heat treatment increased surface hydrophobicity, induced SPI aggregation via hydrophobic interaction and disulfide bonds, and formed soluble aggregates of different sizes. Heating of 5% SPI always promoted large-size aggregate (LA; >1000 kDa) formation irrespective of temperature, whereas the aggregate size distribution in 2% SPI was temperature dependent: the LA fraction progressively rose with temperature (80→90→100 °C), corresponding to the attenuation of medium-size aggregates (MA; 670 to 1000 kDa) initially abundant at 80 °C. Heated SPI with abundant LA (>50%) promoted foam stability. LA also exhibited excellent emulsifying activity and stabilized emulsions by promoting the formation of small oil droplets covered with a thick interfacial protein layer. However, despite a similar influence on emulsion stability, MA enhanced foaming capacity but were less capable of stabilizing emulsions than LA. The functionality variation between heated SPI samples is clearly related to the distribution of aggregates that differ in molecular size and surface activity. The findings may encourage further research to develop functional SPI aggregates for various commercial applications. © 2015 Institute of Food Technologists®

  15. Palatability of water-soluble extracts of protein sources and replacement of fishmeal by a selected mixture of protein sources for juvenile turbot ( Scophthalmus maximus)

    Science.gov (United States)

    Dong, Chun; He, Gen; Mai, Kangsen; Zhou, Huihui; Xu, Wei

    2016-06-01

    Poor palatability is a limiting factor for replacing fishmeal with other protein sources in aquaculture. The water-soluble molecules with low molecular weights are the major determinants of the palatability of diets. The present study was conducted to investigate the palatability of water-soluble extracts from single protein source (single extract pellets) and the mixture of these extracts with different proportions (blended extract pellets) in juvenile turbot ( Scophthalmus maximus). Then according to the palatability of blended extract pellets, an optimal mixture proportion was selected, and a new protein source made from raw protein materials with the selected proportion was formulated to replace fishmeal. Summarily, the palatability of single extract pellets for turbot was descendent from fishmeal to pet-food grade poultry by-product meal, wheat gluten meal, soybean meal, peanut meal, meat and bone meal, and corn gluten meal. Subsequently, according to the palatability of single extract pellets, 52 kinds of blended extract pellets were designed to test their palatability. The results showed that the pellets presented remarkably different palatability, and the optimal one was diet 52 (wheat gluten meal: pet-food grade poultry by-product meal: meat and bone meal: corn gluten meal = 1:6:1:2). The highest ingestion ratio (the number of pellets ingested/the number of pellets fed) was 0.73 ± 0.03, which was observed in Diet 52. Then five isonitrogenous (52% crude protein) and isocaloric (20 kJ g-1 gross energy) diets were formulated by replacing 0 (control), 35%, 50%, 65% and 80% of fishmeal with No.52 blending proportion. After a 10-weeks feeding trial, a consistent feed intake was found among all replacement treatments. Replacement level of fishmeal up to 35% did not significantly influence final body weight, specific growth rate, feed efficiency ratio, and protein efficiency ratio of turbot. Therefore, the water-soluble extracts of protein sources play an

  16. Sialic Acid Binding Properties of Soluble Coronavirus Spike (S1 Proteins: Differences between Infectious Bronchitis Virus and Transmissible Gastroenteritis Virus

    Directory of Open Access Journals (Sweden)

    Christine Winter

    2013-07-01

    Full Text Available The spike proteins of a number of coronaviruses are able to bind to sialic acids present on the cell surface. The importance of this sialic acid binding ability during infection is, however, quite different. We compared the spike protein of transmissible gastroenteritis virus (TGEV and the spike protein of infectious bronchitis virus (IBV. Whereas sialic acid is the only receptor determinant known so far for IBV, TGEV requires interaction with its receptor aminopeptidase N to initiate infection of cells. Binding tests with soluble spike proteins carrying an IgG Fc-tag revealed pronounced differences between these two viral proteins. Binding of the IBV spike protein to host cells was in all experiments sialic acid dependent, whereas the soluble TGEV spike showed binding to APN but had no detectable sialic acid binding activity. Our results underline the different ways in which binding to sialoglycoconjugates is mediated by coronavirus spike proteins.

  17. Infrared dust emission from globular clusters

    International Nuclear Information System (INIS)

    Angeletti, L.; Capuzzo-Dolcetta, R.; Giannone, P.; Blanco, A.; Bussoletti, E.

    1982-01-01

    The implications of the presence of a central cloud in the cores of globular clusters were investigated recently. A possible mechanism of confinement of dust in the central region of our cluster models was also explored. The grain temperature and infrared emission have now been computed for rather realistic grain compositions. The grain components were assumed to be graphite and/or silicates. The central clouds turned out to be roughly isothermal. The wavelengths of maximum emission came out to be larger than 20 μm in all studied cases. An application of the theoretical results to five globular clusters showed that the predictable infrared emission for 47 Tuc, M4 and M22 should be detectable by means of present instrumentation aboard flying platforms. (author)

  18. Infrared dust emission from globular clusters

    Energy Technology Data Exchange (ETDEWEB)

    Angeletti, L; Capuzzo-Dolcetta, R; Giannone, P. (Rome Univ. (Italy). Osservatorio Astronomico); Blanco, A; Bussoletti, E [Lecce Univ. (Italy). Ist. di Fisica

    1982-05-01

    The implications of the presence of a central cloud in the cores of globular clusters were investigated recently. A possible mechanism of confinement of dust in the central region of our cluster models was also explored. The grain temperature and infrared emission have now been computed for rather realistic grain compositions. The grain components were assumed to be graphite and/or silicates. The central clouds turned out to be roughly isothermal. The wavelengths of maximum emission came out to be larger than 20 ..mu..m in all studied cases. An application of the theoretical results to five globular clusters showed that the predictable infrared emission for 47 Tuc, M4 and M22 should be detectable by means of present instrumentation aboard flying platforms.

  19. Effect of enzymatic treatment of extracted sunflower proteins on solubility, amino acid composition, and surface activity.

    Science.gov (United States)

    Conde, José Miñones; Escobar, María del Mar Yust; Pedroche Jiménez, Justo J; Rodríguez, Francisco Millán; Rodríguez Patino, Juan M

    2005-10-05

    Industrial proteins from agriculture of either animal or vegetable origin, including their peptide derivatives, are of great importance, from the qualitative and quantitative point of view, in food formulations (emulsions and foams). A fundamental understanding of the physical, chemical, and functional properties of these proteins is essential if the performance of proteins in foods is to be improved and if underutilized proteins, such as plant proteins (and their hydrolysates and peptides derivatives), are to be increasingly used in traditional and new processed food products (safe, high-quality, health foods with good nutritional value). In this contribution we have determined the main physicochemical characteristics (solubility, composition, and analysis of amino acids) of a sunflower protein isolate (SPI) and its hydrolysates with low (5.62%), medium (23.5%), and high (46.3%) degrees of hydrolysis. The hydrolysates were obtained by enzymatic treatment with Alcalase 2.4 L for DH 5.62 and 23.5% and with Alcalase 2.4 L and Flavorzyme 1000 MG sequentially for DH 46.3%. The protein concentration dependence on surface pressure (surface pressure isotherm), a measure of the surface activity of the products (SPI and its hydrolysates), was obtained by tensiometry. We have observed that the degree of hydrolysis has an effect on solubility, composition, and content of the amino acids of the SPI and its hydrolysates. The superficial activity and the adsorption efficiency were also affected by the degree of hydrolysis.

  20. Small acid soluble proteins for rapid spore identification.

    Energy Technology Data Exchange (ETDEWEB)

    Branda, Steven S.; Lane, Todd W.; VanderNoot, Victoria A.; Jokerst, Amanda S.

    2006-12-01

    This one year LDRD addressed the problem of rapid characterization of bacterial spores such as those from the genus Bacillus, the group that contains pathogenic spores such as B. anthracis. In this effort we addressed the feasibility of using a proteomics based approach to spore characterization using a subset of conserved spore proteins known as the small acid soluble proteins or SASPs. We proposed developing techniques that built on our previous expertise in microseparations to rapidly characterize or identify spores. An alternative SASP extraction method was developed that was amenable to both the subsequent fluorescent labeling required for laser-induced fluorescence detection and the low ionic strength requirements for isoelectric focusing. For the microseparations, both capillary isoelectric focusing and chip gel electrophoresis were employed. A variety of methods were evaluated to improve the molecular weight resolution for the SASPs, which are in a molecular weight range that is not well resolved by the current methods. Isoelectric focusing was optimized and employed to resolve the SASPs using UV absorbance detection. Proteomic signatures of native wild type Bacillus spores and clones genetically engineered to produce altered SASP patterns were assessed by slab gel electrophoresis, capillary isoelectric focusing with absorbance detection as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection.

  1. Integrated photometry of globular star clusters in the Vilnius system

    International Nuclear Information System (INIS)

    Zdanavichyus, K.V.

    1983-01-01

    Integrated colour indices in the Vilnius photometric system and newly determined colour excesses Esub(B-V) for 39 globular clusters are presented. It is shown that the coincidence of integrated spectral types are not a sufficient criterion for the identity of intrinsic colour indices of globular clusters. Relation of integrated colour indices with the slope of the giant branch S and with the horizontal branch morphological type D is investigated. Integrated colour indices of clusters with a blue horizontal branch show no correlation with either D or S. The increase of colour indices of the clusters of types D >= 4 correlates with the distribution of stars along the horizontal branch. Integrated photometry of globular star clusters in the Vilnius multicoloured photometric system permits to determine their colour excesses from some Q diagrams and normal colour index. Integral normal colour indexes and Q parameters for I globular star clusters of the Mironov group display small changes as compared to clusters of group 2. Colour indexes among star clusters having only red horizontal branches (D=7) change most considerably

  2. Synthesis of Salt Soluble Proteins in Barley. Pulse-Labeling Study of Grain Filling in Liquid-Cultured Detached Spikes

    DEFF Research Database (Denmark)

    Giese, Nanna Henriette; Hejgaard, Jørn

    1984-01-01

    The accumulation of salt-soluble proteins in the endosperm of developing barley (Hordeum vulgare L.) grains was examined. Detached spikes of barley were cultured at different levels of nitrogen nutrition and pulse-labeled with [14C] sucrose at specific times after anthesis. Proteins were extracted...... to increased nitrogen nutrition. Two major components, β-amylase and protein Z in particular, had a synthesis profile almost identical to that of the endosperm storage protein, hordein....

  3. Near infrared photometry of globular clusters

    International Nuclear Information System (INIS)

    Evans, T.L.; Menzies, J.W.

    1977-01-01

    Photographic photometry on the V, Isub(K) system has been obtained for giant stars in the metal-rich globular clusters NGC 5927, 6171, 6352, 6356, 6388, 6522, 6528, 6712 and 6723. Colour-magnitude diagrams are presented. These data, with earlier observations of NGC 104 (47 Tuc), yield new parameters to describe the giant branch. These are the colour of the red variables, represented by their mean colour (V - Isub(K)) 0 or by the colour (V - Isub(K))sub(BO) of the bluest red variable on the giant branch of a cluster, and ΔV' which is the magnitude difference between the horizontal branch and the highest point on the giant branch. The latter is independent of reddening, since the giant branch of the most metal-rich clusters passes through a maximum in the V, V - Isub(K) plane. These parameters are correlated with the metal content, deduced from integrated photometry: the red variables are redder and the giant branch fainter the higher the metal content. Comparison with theoretical evolutionary tracks suggests that the range in metal content of these clusters is at most a factor of 10, the most metal-rich clusters possibly approaching the solar value. The cluster giant branches and those of open clusters, groups and field stars of the old disk population are compared. The assumption that all the globular clusters have an absolute magnitude on the horizontal branch of Msub(v) = + 0.9, as found recently for 47 Tuc, gives good agreement between the magnitudes of giant stars in the most metal rich of the globular clusters and those of field stars deduced from statistical parallaxes and moving group parallaxes. The values of the parameters ΔV' and (V - Isub(k))sub(BO) also approach those in the moving groups. The globular clusters have a longer horizontal branch, however, and the subgiants are bluer even when the values of ) 7Fe/H{ appear to be the same. (author). )

  4. Spectroscopic studies on the interaction of a water-soluble cationic porphyrin with proteins

    Science.gov (United States)

    Ma, Hong-Min; Chen, Xin; Zhang, Nuo; Han, Yan-Yan; Wu, Dan; Du, Bin; Wei, Qin

    2009-04-01

    The interaction of a water-soluble cationic porphyrin, meso-tetrakis (4- N, N, N-trimethylanilinium) porphyrin (TMAP), with two proteins, bovine serum albumin (BSA) and human serum albumin (HSA), was studied by UV-vis absorption spectroscopy, fluorescence spectroscopy, fluorescence anisotropy and synchronous fluorescence spectroscopy at neutral aqueous solutions. Free base TMAP bound to proteins as monomers and no aggregation was observed. The binding of TMAP quenched the fluorescence of the protein. On the contrary, the fluorescence of TMAP was enhanced and the fluorescence anisotropy increased due to the binding. The direct static binding mechanism could account for the quenching by TMAP and the binding constants were calculated. TMAP showed a higher quenching efficiency and binding constant of HSA than BSA. The binding of TMAP had no obvious effect on the molecular conformation of the protein. There was only one binding site for TMAP and it was located on the surface of the protein molecule. Electrostatic force played an important role in the binding due to the opposite charges on porphyrin and the proteins.

  5. On the evolution of globular clusters and the origin of galactic halo stars

    International Nuclear Information System (INIS)

    Surdin, V.G.

    1978-01-01

    Evolution of globular clusters of galactic halo is considered. It is shown that evolution of massive globular clusters with a greater degree of probability takes place under the effect of dynamic friction, which brings about the cluster fall on the center of galactic and their destruction by tidal forces. Evolution of small massive cluster takes place under the effect of dissipation. All the other reasons, causing the destruction of globular clusters (gravitational tidal forces, mutual cluster collision, outflow of gas from red gigant atmospheres, the change of the radius of the cluster orbit at the expense of the change of the galaxy mass inside the cluster orbit) play a secondary role. The whole mass of the stars lost by globular clusters does not exceed 10 7 M sun. It is concluded that the origin of the star population of galactic halo field can not be explained by destruction of already formed only astral globular clusters

  6. Application of a PEG precipitation method for solubility screening: A tool for developing high protein concentration formulations

    OpenAIRE

    Li, Li; Kantor, Angela; Warne, Nicholas

    2013-01-01

    Previous publications demonstrated that the extrapolated solubility by polyethylene glycol (PEG) precipitation method (Middaugh et al., J Biol Chem 1979; 254:367–370; Juckes, Biochim Biophys Acta 1971; 229:535–546; Foster et al., Biochim Biophys Acta 1973; 317:505; Mahadevan and Hall, AIChE J 1990; 36:1517–1528; Stevenson and Hageman, Pharm Res 1995; 12:1671–1676) has a strong correlation to experimentally measured solubility of proteins. Here, we explored the utility of extrapolated solubili...

  7. Analytical Solution for Stellar Density in Globular Clusters MA Sharaf

    Indian Academy of Sciences (India)

    Introduction. A globular cluster is a spherical collection of stars that orbits a galactic core as a satellite. They are generally composed of hundreds of thousands of low-metal, old stars. The types of stars found in a globular cluster are similar to those in the bulge of a spiral galaxy but confined to a volume of only a few cubic ...

  8. Interplay between Peptide Bond Geometrical Parameters in Nonglobular Structural Contexts

    OpenAIRE

    Esposito, Luciana; Balasco, Nicole; De Simone, Alfonso; Berisio, Rita; Vitagliano, Luigi

    2013-01-01

    Several investigations performed in the last two decades have unveiled that geometrical parameters of protein backbone show a remarkable variability. Although these studies have provided interesting insights into one of the basic aspects of protein structure, they have been conducted on globular and water-soluble proteins. We report here a detailed analysis of backbone geometrical parameters in nonglobular proteins/peptides. We considered membrane proteins and two distinct fibrous systems (am...

  9. Predicting protein folding rate change upon point mutation using residue-level coevolutionary information.

    Science.gov (United States)

    Mallik, Saurav; Das, Smita; Kundu, Sudip

    2016-01-01

    Change in folding kinetics of globular proteins upon point mutation is crucial to a wide spectrum of biological research, such as protein misfolding, toxicity, and aggregations. Here we seek to address whether residue-level coevolutionary information of globular proteins can be informative to folding rate changes upon point mutations. Generating residue-level coevolutionary networks of globular proteins, we analyze three parameters: relative coevolution order (rCEO), network density (ND), and characteristic path length (CPL). A point mutation is considered to be equivalent to a node deletion of this network and respective percentage changes in rCEO, ND, CPL are found linearly correlated (0.84, 0.73, and -0.61, respectively) with experimental folding rate changes. The three parameters predict the folding rate change upon a point mutation with 0.031, 0.045, and 0.059 standard errors, respectively. © 2015 Wiley Periodicals, Inc.

  10. A New Globular Cluster in the Area of VVVX

    Science.gov (United States)

    Bica, E.; Minniti, D.; Bonatto, C.; Hempel, M.

    2018-06-01

    We communicate the discovery of a new globular cluster in the Galaxy that was first detected on WISE/2MASS images and is now confirmed with VVVX photometry. It is a Palomar-like cluster projected at ℓ = 359.15°, b = 5.73°, and may be related to the bulge. We derive an absolute magnitude of MV ≈ -3.3, thus being an underluminous globular cluster. Our analyses provide a reddening of E(B - V) = 1.08 ± 0.18 and a distance to the Sun d⊙ = 6.3 ± 1 kpc, which implies a current position in the bulge volume. The estimated metallicity is [Fe/H] = -1.5 ± 0.25. It adds to the recently discovered faint globular cluster (Minniti 22) and candidates found with VVV, building up expectations of ≈50 globular clusters yet to be discovered in the bulge. We also communicate the discovery of an old open cluster in the same VVVX tile as the globular cluster. The VVVX photometry provided E(B - V) = 0.62 ± 0.1, d⊙ = 7.6 ± 1 kpc, and an age of 1.5 ± 0.3 Gyr. With a height from the plane of ≈0.8 kpc, it adds to nine Gyr-class clusters recently discovered within 0.8 ⩽ Z ⩽ 2.2 kpc, as recently probed in the single VVV tile b201. We suggest that these findings may be disclosing the thick disk at the bulge, which so far has no open cluster counterpart, and hardly any individual star. Thus, the VVV and VVVX surveys are opening new windows for follow-up studies, to employ present and future generations of large aperture telescopes.

  11. Systematic main sequence photometry of globular cluster stars for age determination

    International Nuclear Information System (INIS)

    Alcaino, G.; Liller, W.

    1984-01-01

    The individual photometric study of the coeval stars in globular clusters presents one of the best observational tests of the stellar evolution theory. Our own globular cluster system provides fundamental clues to the dynamical and chemical evolutionary history of the galaxy, and the study of their ages give a lower limit to the age of the galaxy as well as to that of the universe. The authors have undertaken a systematic research program, and discuss the ages deduced by fitting main sequence photometry to theoretical isochrones of six galactic globular clusters: M4, M22, M30, NGC 288, NGC 3201 and NGC 6397. (Auth.)

  12. EVIDENCE FOR AN ACCRETION ORIGIN FOR THE OUTER HALO GLOBULAR CLUSTER SYSTEM OF M31

    International Nuclear Information System (INIS)

    Mackey, A. D.; Huxor, A. P.; Ferguson, A. M. N.; Irwin, M. J.; Chapman, S. C.; Tanvir, N. R.; McConnachie, A. W.; Ibata, R. A.; Lewis, G. F.

    2010-01-01

    We use a sample of newly discovered globular clusters from the Pan-Andromeda Archaeological Survey (PAndAS) in combination with previously cataloged objects to map the spatial distribution of globular clusters in the M31 halo. At projected radii beyond ∼30 kpc, where large coherent stellar streams are readily distinguished in the field, there is a striking correlation between these features and the positions of the globular clusters. Adopting a simple Monte Carlo approach, we test the significance of this association by computing the probability that it could be due to the chance alignment of globular clusters smoothly distributed in the M31 halo. We find that the likelihood of this possibility is low, below 1%, and conclude that the observed spatial coincidence between globular clusters and multiple tidal debris streams in the outer halo of M31 reflects a genuine physical association. Our results imply that the majority of the remote globular cluster system of M31 has been assembled as a consequence of the accretion of cluster-bearing satellite galaxies. This constitutes the most direct evidence to date that the outer halo globular cluster populations in some galaxies are largely accreted.

  13. The central globular domain of the nucleocapsid protein of human immunodeficiency virus type 1 is critical for virion structure and infectivity.

    Science.gov (United States)

    Ottmann, M; Gabus, C; Darlix, J L

    1995-03-01

    The nucleocapsid protein NCp7 of human immunodeficiency virus type 1 (HIV-1) is a 72-amino-acid peptide containing two CCHC-type zinc fingers linked by a short basic sequence, 29RAPRKKG35, which is conserved in HIV-1 and simian immunodeficiency virus. The complete three-dimensional structure of NCp7 has been determined by 1H-nuclear magnetic resonance spectroscopy (N. Morellet, H. de Rocquigny, Y. Mely, N. Jullian, H. Demene, M. Ottmann, D. Gerard, J. L. Darlix, M. C. Fournié-Zaluski, and B. P. Roques, J. Mol. Biol. 235:287-301, 1994) and revealed a central globular domain where the two zinc fingers are brought in close proximity by the RAPRKKG linker. To examine the role of this globular structure and more precisely of the RAPRKKG linker in virion structure and infectivity, we generated HIV-1 DNA mutants in the RAPRKK sequence of NCp7 and analyzed the mutant virions produced by transfected cells. Mutations that probably alter the structure of NCp7 structure led to the formation of very poorly infectious virus (A30P) or noninfectious virus (P31L and R32G). In addition, the P31L mutant did not contain detectable amounts of reverse transcriptase and had an immature core morphology, as determined by electron microscopy. On the other hand, mutations changing the basic nature of NCp7 had poor effect. R29S had a wild-type phenotype, and the replacement of 32RKK34 by SSS (S3 mutant) resulted in a decrease by no more than 100-fold of the virus titer. These results clearly show that the RAPRKKG linker contains residues that are critical for virion structure and infectivity.

  14. Globular and disordered-the non-identical twins in protein-protein interactions

    DEFF Research Database (Denmark)

    Teilum, Kaare; Olsen, Johan Gotthardt; Kragelund, Birthe Brandt

    2015-01-01

    as a strong determinant for their function. This has fostered the notion that IDP's bind with low affinity but high specificity. Here we have analyzed available detailed thermodynamic data for protein-protein interactions to put to the test if the thermodynamic profiles of IDP interactions differ from those...... of other protein-protein interactions. We find that ordered proteins and the disordered ones act as non-identical twins operating by similar principles but where the disordered proteins complexes are on average less stable by 2.5 kcal mol(-1)....

  15. Glucose, fructose and sucrose increase the solubility of protein-tannin complexes and at high concentration, glucose and sucrose interfere with bisulphite bleaching of wine pigments.

    Science.gov (United States)

    Harbertson, James F; Yuan, Chunlong; Mireles, Maria S; Hanlin, Rachel L; Downey, Mark O

    2013-05-01

    Wines were modified with increasing sugar concentrations and decreasing tannin concentrations and analysed by a combination of protein precipitation and bisulphite bleaching. Increasing sugar concentration decreased the precipitation of tannin and protein-precipitable polymeric pigments (PPP). The use of a hydrogen bond disruptor (urea) to reduce protein-tannin and protein-pigment complex formation showed that the effect of sugar concentration occurred by increasing the solubility of the tannin-protein complex, not by interfering with protein-tannin complex formation. By increasing the solubility of pigment-protein complexes, non-protein-precipitable polymeric pigments (nPPP) appeared to increase. There was also an increase in total polymeric pigments at each tannin concentration with increasing glucose and sucrose concentration, indicating that sugar concentration might also affect bisulphite bleaching of wine pigments. While a significant effect of sugar concentration on tannin-protein complex solubility was observed, these effects were greatest at sugar concentrations far in excess of normal wine making conditions. Under normal wine making conditions, sugar concentration will have a negligible effect on protein-precipitable tannin, PPP and nPPP concentrations. Copyright © 2012 Elsevier Ltd. All rights reserved.

  16. Bacterial CpG-DNA activates dendritic cells in vivo: T helper cell-independent cytotoxic T cell responses to soluble proteins.

    Science.gov (United States)

    Sparwasser, T; Vabulas, R M; Villmow, B; Lipford, G B; Wagner, H

    2000-12-01

    Receptors for conserved molecular patterns associated with microbial pathogens induce synthesis of co-stimulatory molecules and cytokines in immature dendritic cells (DC), as do antigen-reactive CD4 T helper cells via CD40 signaling. Once activated, antigen-presenting DC may activate CD8 T cell responses in a T helper cell-independent fashion. Using immunostimulatory CpG-oligonucleotides (ODN) mimicking bacterial CpG-DNA, we tested whether CpG-DNA bypasses the need for T helper cells in CTL responses towards proteins by directly activating antigen-presenting DC to transit into professional APC. We describe that immature DC in situ constitutively process soluble proteins and generate CD8 T cell determinants yet CD8 T cell responses remain abortive. Induction of primary antigen-specific CD8 cytotoxic T lymphocyte (CTL)-mediated responses becomes initiated in wild-type as well as T helper cell-deficient mice, provided soluble protein and CpG-ODN are draining into the same lymph node. Specifically we show that CpG-ODN trigger antigen-presenting immature DC within the draining lymph node to acutely up-regulate co-stimulatory molecules and produce IL-12. These results provide new insights for generating in vivo efficient CTL responses to soluble proteins which may influence vaccination strategies.

  17. Seeing deconvolution of globular clusters in M31

    International Nuclear Information System (INIS)

    Bendinelli, O.; Zavatti, F.; Parmeggiani, G.; Djorgovski, S.

    1990-01-01

    The morphology of six M31 globular clusters is examined using seeing-deconvolved CCD images. The deconvolution techniques developed by Bendinelli (1989) are reviewed and applied to the M31 globular clusters to demonstrate the methodology. It is found that the effective resolution limit of the method is about 0.1-0.3 arcsec for CCD images obtained in FWHM = 1 arcsec seeing, and sampling of 0.3 arcsec/pixel. Also, the robustness of the method is discussed. The implications of the technique for future studies using data from the Hubble Space Telescope are considered. 68 refs

  18. Heat-induced alterations in cashew allergen solubility and IgE binding

    Directory of Open Access Journals (Sweden)

    Christopher P. Mattison

    Full Text Available Cashew nuts are an increasingly common cause of food allergy. We compare the soluble protein profile of cashew nuts following heating. SDS-PAGE indicate that heating can alter the solubility of cashew nut proteins. The 11S legumin, Ana o 2, dominates the soluble protein content in ready to eat and mildly heated cashew nuts. However, we found that in dark-roasted cashew nuts, the soluble protein profile shifts and the 2S albumin Ana o 3 composes up to 40% of the soluble protein. Analysis of trypsin-treated extracts by LC/MS/MS indicate changes in the relative number and intensity of peptides. The relative cumulative intensity of the 5 most commonly observed Ana o 1 and 2 peptides are altered by heating, while those of the 5 most commonly observed Ana o 3 peptides remaine relatively constant. ELISA experiments indicate that there is a decrease in rabbit IgG and human serum IgE binding to soluble cashew proteins following heating. Our findings indicate that heating can alter the solubility of cashew allergens, resulting in altered IgE binding. Our results support the use of both Ana o 2 and Ana o 3 as potential cashew allergen diagnostic targets. Keywords: Cashew nut, Food allergy, Immunoglobulin E, Mass-spectrometry, Peptide, Solubility

  19. Statistical mechanics of protein solutions

    NARCIS (Netherlands)

    Prinsen, P.

    2007-01-01

    We study theoretically thermodynamic properties of spherical globular proteins in aqueous solution with added monovalent salt. We show how one can determine an effective interaction potential between the proteins from experimental data as a function of salt concentration and we apply this to the

  20. Observations of CO and OI in stars in globular clusters

    International Nuclear Information System (INIS)

    Wallerstein, G.; Pilachowski, C.

    1978-01-01

    Since studies at classification dispersion and early analyses of high dispersion spectra have yielded little quantitative data on the abundances of C, N, and O in globular clusters the authors have been endeavoring to establish their abundances in stars in several clusters. The problem has been approached in two ways, by observing the 2.3 micron CO bands and the 6300 A [OI] line in individual stars in globular clusters. (Auth.)

  1. Plasma soluble prion protein, a potential biomarker for sport-related concussions: a pilot study.

    Science.gov (United States)

    Pham, Nam; Akonasu, Hungbo; Shishkin, Rhonda; Taghibiglou, Changiz

    2015-01-01

    Sport-related mild traumatic brain injury (mTBI) or concussion is a significant health concern to athletes with potential long-term consequences. The diagnosis of sport concussion and return to sport decision making is one of the greatest challenges facing health care clinicians working in sports. Blood biomarkers have recently demonstrated their potential in assisting the detection of brain injury particularly, in those cases with no obvious physical injury. We have recently discovered plasma soluble cellular prion protein (PrP(C)) as a potential reliable biomarker for blast induced TBI (bTBI) in a rodent animal model. In order to explore the application of this novel TBI biomarker to sport-related concussion, we conducted a pilot study at the University of Saskatchewan (U of S) by recruiting athlete and non-athlete 18 to 30 year-old students. Using a modified quantitative ELISA method, we first established normal values for the plasma soluble PrP(C) in male and female students. The measured plasma soluble PrP(C) in confirmed concussion cases demonstrated a significant elevation of this analyte in post-concussion samples. Data collected from our pilot study indicates that the plasma soluble PrP(C) is a potential biomarker for sport-related concussion, which may be further developed into a clinical diagnostic tool to assist clinicians in the assessment of sport concussion and return-to-play decision making.

  2. The gamma-ray pulsar population of globular clusters: implications for the GeV excess

    Energy Technology Data Exchange (ETDEWEB)

    Hooper, Dan [Fermi National Accelerator Laboratory, Center for Particle Astrophysics, Batavia, IL 60510 (United States); Linden, Tim, E-mail: dhooper@fnal.gov, E-mail: linden.70@osu.edu [Ohio State University, Center for Cosmology and AstroParticle Physcis (CCAPP), Columbus, OH 43210 (United States)

    2016-08-01

    It has been suggested that the GeV excess, observed from the region surrounding the Galactic Center, might originate from a population of millisecond pulsars that formed in globular clusters. With this in mind, we employ the publicly available Fermi data to study the gamma-ray emission from 157 globular clusters, identifying a statistically significant signal from 25 of these sources (ten of which are not found in existing gamma-ray catalogs). We combine these observations with the predicted pulsar formation rate based on the stellar encounter rate of each globular cluster to constrain the gamma-ray luminosity function of millisecond pulsars in the Milky Way's globular cluster system. We find that this pulsar population exhibits a luminosity function that is quite similar to those millisecond pulsars observed in the field of the Milky Way (i.e. the thick disk). After pulsars are expelled from a globular cluster, however, they continue to lose rotational kinetic energy and become less luminous, causing their luminosity function to depart from the steady-state distribution. Using this luminosity function and a model for the globular cluster disruption rate, we show that millisecond pulsars born in globular clusters can account for only a few percent or less of the observed GeV excess. Among other challenges, scenarios in which the entire GeV excess is generated from such pulsars are in conflict with the observed mass of the Milky Way's Central Stellar Cluster.

  3. A Science Portal and Archive for Extragalactic Globular Cluster Systems Data

    Science.gov (United States)

    Young, Michael; Rhode, Katherine L.; Gopu, Arvind

    2015-01-01

    For several years we have been carrying out a wide-field imaging survey of the globular cluster populations of a sample of giant spiral, S0, and elliptical galaxies with distances of ~10-30 Mpc. We use mosaic CCD cameras on the WIYN 3.5-m and Kitt Peak 4-m telescopes to acquire deep BVR imaging of each galaxy and then analyze the data to derive global properties of the globular cluster system. In addition to measuring the total numbers, specific frequencies, spatial distributions, and color distributions for the globular cluster populations, we have produced deep, high-quality images and lists of tens to thousands of globular cluster candidates for the ~40 galaxies included in the survey.With the survey nearing completion, we have been exploring how to efficiently disseminate not only the overall results, but also all of the relevant data products, to the astronomical community. Here we present our solution: a scientific portal and archive for extragalactic globular cluster systems data. With a modern and intuitive web interface built on the same framework as the WIYN One Degree Imager Portal, Pipeline, and Archive (ODI-PPA), our system will provide public access to the survey results and the final stacked mosaic images of the target galaxies. In addition, the astrometric and photometric data for thousands of identified globular cluster candidates, as well as for all point sources detected in each field, will be indexed and searchable. Where available, spectroscopic follow-up data will be paired with the candidates. Advanced imaging tools will enable users to overlay the cluster candidates and other sources on the mosaic images within the web interface, while metadata charting tools will allow users to rapidly and seamlessly plot the survey results for each galaxy and the data for hundreds of thousands of individual sources. Finally, we will appeal to other researchers with similar data products and work toward making our portal a central repository for data

  4. Effects of non-covalent interactions with 5-O-caffeoylquinic acid (chlorogenic acid) on the heat denaturation and solubility of globular proteins

    NARCIS (Netherlands)

    Prigent, S.V.E.; Gruppen, H.; Visser, A.J.W.G.; Koningsveld, G.A. van; Jong, G.A.H. de; Voragen, A.G.J.

    2003-01-01

    The non-covalent interactions between the monomeric phenolic compound chlorogenic acid (5-CQA) and bovine serum albumin (BSA), lysozyme, and α-lactalbumin were characterized, and their effect on protein properties was examined. 5-CQA had a low affinity for all three proteins, and these interactions

  5. Why fibrous proteins are romantic.

    Science.gov (United States)

    Cohen, C

    1998-01-01

    Here I give a personal account of the great history of fibrous protein structure. I describe how Astbury first recognized the essential simplicity of fibrous proteins and their paradigmatic role in protein structure. The poor diffraction patterns yielded by these proteins were then deciphered by Pauling, Crick, Ramachandran and others (in part by model building) to reveal alpha-helical coiled coils, beta-sheets, and the collagen triple helical coiled coil-all characterized by different local sequence periodicities. Longer-range sequence periodicities (or "magic numbers") present in diverse fibrous proteins, such as collagen, tropomyosin, paramyosin, myosin, and were then shown to account for the characteristic axial repeats observed in filaments of these proteins. More recently, analysis of fibrous protein structure has been extended in many cases to atomic resolution, and some systems, such as "leucine zippers," are providing a deeper understanding of protein design than similar studies of globular proteins. In the last sections, I provide some dramatic examples of fibrous protein dynamics. One example is the so-called "spring-loaded" mechanism for viral fusion by the hemagglutinin protein of influenza. Another is the possible conformational changes in prion proteins, implicated in "mad cow disease," which may be related to similar transitions in a variety of globular and fibrous proteins. Copyright 1998 Academic Press.

  6. Two stellar-mass black holes in the globular cluster M22.

    Science.gov (United States)

    Strader, Jay; Chomiuk, Laura; Maccarone, Thomas J; Miller-Jones, James C A; Seth, Anil C

    2012-10-04

    Hundreds of stellar-mass black holes probably form in a typical globular star cluster, with all but one predicted to be ejected through dynamical interactions. Some observational support for this idea is provided by the lack of X-ray-emitting binary stars comprising one black hole and one other star ('black-hole/X-ray binaries') in Milky Way globular clusters, even though many neutron-star/X-ray binaries are known. Although a few black holes have been seen in globular clusters around other galaxies, the masses of these cannot be determined, and some may be intermediate-mass black holes that form through exotic mechanisms. Here we report the presence of two flat-spectrum radio sources in the Milky Way globular cluster M22, and we argue that these objects are black holes of stellar mass (each ∼10-20 times more massive than the Sun) that are accreting matter. We find a high ratio of radio-to-X-ray flux for these black holes, consistent with the larger predicted masses of black holes in globular clusters compared to those outside. The identification of two black holes in one cluster shows that ejection of black holes is not as efficient as predicted by most models, and we argue that M22 may contain a total population of ∼5-100 black holes. The large core radius of M22 could arise from heating produced by the black holes.

  7. Spectroscopic confirmation of the first symbiotic star in a globular cluster

    Science.gov (United States)

    Zurek, David

    2013-10-01

    We have recently discovered an 18-minute period in the ultraviolet of a star in the globular cluster NGC 1851. In the redder optical bands, this star is red and bright, while it shows a clear UV excess relative to other stars at similar positions in the HR diagram. The system is most likely a symbiotic binary, composed of a cool evolved star and a white dwarf, with an 18 minute spin period, accreting the cool star's wind. The binary would be the first such object ever found in a globular cluster, and only the third in the Galaxy where the white dwarf spin period is measured. The only viable alternatives are that the two components are a chance superposition - something with a nontrivial chance of happening in a globular cluster core. In such a case, the 18 minute period would most likely be the spin period of a magnetic white dwarf in an intermediate polar cataclysmic variable {this would be the first confirmed magnetic CV in a globular cluster}, or the orbital period of a double-degenerate AM CVn binary. Each of these three possibilities show unique {and very different} emission line spectra in the blue wavelength range. Two orbits of HST with STIS/G430L will produce a spectrum of sufficient signal-to-noise to distinguish between these 3 scenarios. The result will be an important constraint on N-body models of globular clusters.

  8. The Blue Hook Populations of Massive Globular Clusters

    Science.gov (United States)

    Brown, Thomas

    2006-07-01

    Blue hook stars are a class of hot { 35,000 K} subluminous horizontal branch stars that have been recently discovered using HST ultraviolet images of the globular clusters omega Cen and NGC 2808. These stars occupy a region of the HR diagram that is unexplained by canonical stellar evolution theory. Using new theoretical evolutionary and atmospheric models, we have shown that the blue hook stars are very likely the progeny of stars that undergo extensive internal mixing during a late helium core flash on the white dwarf cooling curve. This "flash mixing" produces an enormous enhancement of the surface helium and carbon abundances, which suppresses the flux in the far ultraviolet. Although flash mixing is more likely to occur in stars that are born with high helium abundances, a high helium abundance, by itself, does not explain the presence of a blue hook population - flash mixing of the envelope is required. We propose ACS ultraviolet {SBC/F150LP and HRC/F250W} observations of the five additional globular clusters for which the presence of blue hook stars is suspected from longer wavelength observations. Like omega Cen and NGC 2808, these five targets are also among the most massive globular clusters, because less massive clusters show no evidence for blue hook stars. Because our targets span 1.5 dex in metallicity, we will be able to test our prediction that flash-mixing should be less drastic in metal-rich blue hook stars. In addition, our observations will test the hypothesis that blue hook stars only form in globular clusters massive enough to retain the helium-enriched ejecta from the first stellar generation. If this hypothesis is correct, then our observations will yield important constraints on the chemical evolution and early formation history in globular clusters, as well as the role of helium self-enrichment in producing blue horizontal branch morphologies and multiple main sequence turnoffs. Finally, our observations will provide new insight into the

  9. Effect of phospholipid, detergent and protein-protein interaction on stability and phosphoenzyme isomerization of soluble sarcoplasmic reticulum Ca-ATPase.

    Science.gov (United States)

    Vilsen, B; Andersen, J P

    1987-12-30

    The purpose of the present study was to elucidate the separate roles of lipid, detergent and protein-protein interaction for stability and catalytic properties of sarcoplasmic reticulum Ca-ATPase solubilized in the non-ionic detergent octa(ethylene glycol) monododecyl ether (C12E8). The use of large-zone high-performance liquid chromatography permitted us to define the self-association state of Ca-ATPase peptide at various detergent, phospholipid and protein concentrations, and also during enzymatic turnover with ATP. Conditions were established for monomerization of Ca-ATPase in the presence of a high concentration of phospholipid relative to detergent. The lipid-saturated monomeric preparation was relatively resistant to inactivation in the absence of Ca2+, whereas delipidated enzyme in monomeric or in oligomeric form was prone to inactivation. Kinetics of phosphoenzyme turnover were examined in the presence and absence of Mg2+. Dephosphorylation rates were sensitive to Mg2+, irrespective of whether the peptide was present in soluble monomeric form or was membrane-bound. C12E8-solubilized monomer without added phospholipid was, however, characterized by a fast initial phase of dephosphorylation in the absence of Mg2+. This was not observed with monomer saturated with phospholipid or with monomer solubilized in myristoylglycerophosphocholine or deoxycholate. The mechanism underlying this difference was shown to be a C12E8-induced acceleration of conversion of ADP-sensitive phosphoenzyme (E1P) to ADP-insensitive phosphoenzyme (E2P). The phosphoenzyme isomerization rate was also found to be enhanced by low-affinity binding of ATP. This was demonstrated both in membrane-bound and in soluble monomeric Ca-ATPase. Our results indicate that a single peptide chain constitutes the target for modulation of phosphoenzyme turnover by Mg2+ and ATP, and that detergent effects, distinct from those arising from disruption of protein-protein contacts, are the major determinants of

  10. Evolution of redback radio pulsars in globular clusters

    Science.gov (United States)

    Benvenuto, O. G.; De Vito, M. A.; Horvath, J. E.

    2017-02-01

    Context. We study the evolution of close binary systems composed of a normal, intermediate mass star and a neutron star considering a chemical composition typical of that present in globular clusters (Z = 0.001). Aims: We look for similarities and differences with respect to solar composition donor stars, which we have extensively studied in the past. As a definite example, we perform an application on one of the redbacks located in a globular cluster. Methods: We performed a detailed grid of models in order to find systems that represent the so-called redback binary radio pulsar systems with donor star masses between 0.6 and 2.0 solar masses and orbital periods in the range 0.2-0.9 d. Results: We find that the evolution of these binary systems is rather similar to those corresponding to solar composition objects, allowing us to account for the occurrence of redbacks in globular clusters, as the main physical ingredient is the irradiation feedback. Redback systems are in the quasi-RLOF state, that is, almost filling their corresponding Roche lobe. During the irradiation cycle the system alternates between semi-detached and detached states. While detached the system appears as a binary millisecond pulsar, called a redback. Circumstellar material, as seen in redbacks, is left behind after the previous semi-detached phase. Conclusions: The evolution of binary radio pulsar systems considering irradiation successfully accounts for, and provides a way for, the occurrence of redback pulsars in low-metallicity environments such as globular clusters. This is the case despite possible effects of the low metal content of the donor star that could drive systems away from redback configuration.

  11. Immunization with influenza virus hemagglutinin globular region containing the receptor-binding pocket.

    Science.gov (United States)

    Jeon, Sung Ho; Arnon, Ruth

    2002-01-01

    The globular region of hemagglutinin (residues 91-261) membrane glycoprotein of influenza virus that encompasses the binding zone to the oligosaccharide receptor of target cells has been cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). This protein segment (denoted HA91-261 peptide) induced significant immune response in mice. The serum antibodies and lung homogenates from the immunized mice cross-reacted with native virus particles. The cellular immunity was manifested by proliferative splenocyte responses and cytokine release indicating T helper type 1 activity. The plasmid DNA containing this segment (denoted pHA91-261) provoked, in addition, a significant cytotoxic T lymphocyte (CTL) response, whereas the HA91-261 protein fragment led to no such response. Both the DNA and the protein fragment of HA91-261 induced significant protection against viral challenge, although the immune response they induce might be along different pathways. Interestingly, the combined DNA priming-protein boosting immunization regimen did not induce protection against viral challenges even though it led to significant humoral immune responses similar to that induced by the peptide vaccine.

  12. Self-consistent field theory of protein adsorption in a non-Gaussian polyelectrolyte brush

    NARCIS (Netherlands)

    Biesheuvel, P.M.; Leermakers, F.A.M.; Stuart, M.A.C.

    2006-01-01

    To describe adsorption of globular protein molecules in a polyelectrolyte brush we use the strong-stretching approximation of the Edwards self-consistent field equation, combined with corrections for a non-Gaussian brush. To describe chemical potentials in this mixture of (globular) species of

  13. Lipid-protein nanodiscs for cell-free production of integral membrane proteins in a soluble and folded state: comparison with detergent micelles, bicelles and liposomes.

    Science.gov (United States)

    Lyukmanova, E N; Shenkarev, Z O; Khabibullina, N F; Kopeina, G S; Shulepko, M A; Paramonov, A S; Mineev, K S; Tikhonov, R V; Shingarova, L N; Petrovskaya, L E; Dolgikh, D A; Arseniev, A S; Kirpichnikov, M P

    2012-03-01

    Production of integral membrane proteins (IMPs) in a folded state is a key prerequisite for their functional and structural studies. In cell-free (CF) expression systems membrane mimicking components could be added to the reaction mixture that promotes IMP production in a soluble form. Here lipid-protein nanodiscs (LPNs) of different lipid compositions (DMPC, DMPG, POPC, POPC/DOPG) have been compared with classical membrane mimicking media such as detergent micelles, lipid/detergent bicelles and liposomes by their ability to support CF synthesis of IMPs in a folded and soluble state. Three model membrane proteins of different topology were used: homodimeric transmembrane (TM) domain of human receptor tyrosine kinase ErbB3 (TM-ErbB3, 1TM); voltage-sensing domain of K(+) channel KvAP (VSD, 4TM); and bacteriorhodopsin from Exiguobacterium sibiricum (ESR, 7TM). Structural and/or functional properties of the synthesized proteins were analyzed. LPNs significantly enhanced synthesis of the IMPs in a soluble form regardless of the lipid composition. A partial disintegration of LPNs composed of unsaturated lipids was observed upon co-translational IMP incorporation. Contrary to detergents the nanodiscs resulted in the synthesis of ~80% active ESR and promoted correct folding of the TM-ErbB3. None of the tested membrane mimetics supported CF synthesis of correctly folded VSD, and the protocol of the domain refolding was developed. The use of LPNs appears to be the most promising approach to CF production of IMPs in a folded state. NMR analysis of (15)N-Ile-TM-ErbB3 co-translationally incorporated into LPNs shows the great prospects of this membrane mimetics for structural studies of IMPs produced by CF systems. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Enhanced Soluble Protein and Biochemical Methane Potential of Apple Biowaste by Different Pretreatment

    Science.gov (United States)

    Tulun, Şevket; Bilgin, Melayib

    2018-05-01

    The purpose of this research is to evaluate the anaerobic digestion of apple pomace waste in terms of pretreatment. In this study, the main pretreatment strategies for apple pomace include: ultrasound (35 and 53 kHz), thermal and chemical (pH 5 and 10). For each pretreatment method four different temperatures are selected as 25, 40, 50, and 60 °C, and operation times are selected as 5th, 15th, 30th, and 45th minutes. The effects on pretreatment were investigated by measuring changes in the soluble protein concentrations of pretreated wastes and the enhanced anaerobic digestion was investigated by using the biochemical methane potential (BMP) assay. The soluble proteins of ultrasonic (35 kHz at 60 °C, 45th min), ultrasonic (53 kHz at 60 °C, 45th min), chemical (pH 5 at 60 °C, 5th min), chemical (pH 10 at 60 °C, 30th min) and thermal chemical (40 °C, 15th min) pretreatment apple pomace were 74.3, 75.6, 48.7, 85.5 and 58.6% higher, respectively. The results indicated that apple pomace treated with 53 kHz at 60 °C, 45th min had the highest biogas yield of 1519 mL CH4/g VSS.day after anaerobic digestion, which was on average 40.9% higher than raw pomace.

  15. Enhanced Soluble Protein and Biochemical Methane Potential of Apple Biowaste by Different Pretreatment

    Science.gov (United States)

    Tulun, Şevket; Bilgin, Melayib

    2018-01-01

    The purpose of this research is to evaluate the anaerobic digestion of apple pomace waste in terms of pretreatment. In this study, the main pretreatment strategies for apple pomace include: ultrasound (35 and 53 kHz), thermal and chemical (pH 5 and 10). For each pretreatment method four different temperatures are selected as 25, 40, 50, and 60 °C, and operation times are selected as 5th, 15th, 30th, and 45th minutes. The effects on pretreatment were investigated by measuring changes in the soluble protein concentrations of pretreated wastes and the enhanced anaerobic digestion was investigated by using the biochemical methane potential (BMP) assay. The soluble proteins of ultrasonic (35 kHz at 60 °C, 45th min), ultrasonic (53 kHz at 60 °C, 45th min), chemical (pH 5 at 60 °C, 5th min), chemical (pH 10 at 60 °C, 30th min) and thermal chemical (40 °C, 15th min) pretreatment apple pomace were 74.3, 75.6, 48.7, 85.5 and 58.6% higher, respectively. The results indicated that apple pomace treated with 53 kHz at 60 °C, 45th min had the highest biogas yield of 1519 mL CH4/g VSS.day after anaerobic digestion, which was on average 40.9% higher than raw pomace.

  16. The Most Massive Star Clusters: Supermassive Globular Clusters or Dwarf Galaxy Nuclei?

    Science.gov (United States)

    Harris, William

    2004-07-01

    Evidence is mounting that the most massive globular clusters, such as Omega Centauri and M31-G1, may be related to the recently discovered "Ultra-Compact Dwarfs" and the dense nuclei of dE, N galaxies. However, no systematic imaging investigation of these supermassive globular clusters - at the level of Omega Cen and beyond - has been done, and we do not know what fraction of them might bear the signatures {such as large effective radii or tidal tails} of having originated as dE nuclei. We propose to use the ACS/WFC to obtain deep images of 18 such clusters in NGC 5128 and M31, the two nearest rich globular cluster systems. These globulars are the richest star clusters that can be found in nature, the biggest of them reaching 10^7 Solar masses, and they are likely to represent the results of star formation under the densest and most extreme conditions known. Using the profiles of the clusters including their faint outer envelopes, we will carry out state-of-the-art dynamical modelling of their structures, and look for any clear evidence which would indicate that they are associated with stripped satellites. This study will build on our previous work with STIS and WFPC2 imaging designed to study the 'Fundamental Plane' of globular clusters. When our new work is combined with Archival WFPC2, STIS, and ACS material, we will also be able to construct the definitive mapping of the Fundamental Plane of globular clusters at its uppermost mass range, and confirm whether or not the UCD and dE, N objects occupy a different structural parameter space.

  17. Binding of human collectins (SP-A and MBP) to influenza virus.

    OpenAIRE

    Malhotra, R; Haurum, J S; Thiel, S; Sim, R B

    1994-01-01

    Collectins are a group of soluble proteins each of which has collagenous domains and non-collagenous globular domains, the latter containing the consensus residues found in C-type lectins. Members of the collectin family are the serum proteins mannan-binding protein (MBP), conglutinin, CL-43, and the lung-associated proteins surfactant protein A (SP-A) and surfactant protein D (SP-D). MBP and conglutinin have been shown previously to bind to influenza viruses and to inhibit the infectivity an...

  18. Wheat germ cell-free expression: Two detergents with a low critical micelle concentration allow for production of soluble HCV membrane proteins.

    Science.gov (United States)

    Fogeron, Marie-Laure; Badillo, Aurélie; Jirasko, Vlastimil; Gouttenoire, Jérôme; Paul, David; Lancien, Loick; Moradpour, Darius; Bartenschlager, Ralf; Meier, Beat H; Penin, François; Böckmann, Anja

    2015-01-01

    Membrane proteins are notoriously difficult to express in a soluble form. Here, we use wheat germ cell-free expression in the presence of various detergents to produce the non-structural membrane proteins 2, 4B and 5A of the hepatitis C virus (HCV). We show that lauryl maltose neopentyl glycol (MNG-3) and dodecyl octaethylene glycol ether (C12E8) detergents can yield essentially soluble membrane proteins at detergent concentrations that do not inhibit the cell-free reaction. This finding can be explained by the low critical micelle concentration (CMC) of these detergents, which keeps the monomer concentrations low while at the same time providing the necessary excess of detergent concentration above CMC required for full target protein solubilization. We estimate that a tenfold excess of detergent micelles with respect to the protein concentration is sufficient for solubilization, a number that we propose as a guideline for detergent screening assays. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Design of Fusion Proteins for Efficient and Soluble Production of Immunogenic Ebola Virus Glycoprotein in Escherichia coli.

    Science.gov (United States)

    Ji, Yang; Lu, Yuan; Yan, Yishu; Liu, Xinxin; Su, Nan; Zhang, Chong; Bi, Shengli; Xing, Xin-Hui

    2018-03-03

    The Ebola hemorrhagic fever caused by Ebola virus is an extremely dangerous disease, and effective therapeutic agents are still lacking. Platforms for the efficient production of vaccines are crucial to ensure quick response against an Ebola virus outbreak. Ebola virus glycoprotein (EbolaGP) on the virion surface is responsible for membrane binding and virus entry, thus becoming the key target for vaccine development. However, heterologous expression of this protein still faces engineering challenges such as low production levels and insoluble aggregation. Here, the authors design and compare various fusion strategies, attaching great importance to the solubility-enhancing effect, and tag removal process. It is found that a C-terminal intein-based tag greatly enhances the solubility of EbolaGP and allows one-step chromatographic purification of the untagged EbolaGP through thiol-catalyzed self-cleavage. The purified untagged EbolaGP alone or with Freund's adjuvant are highly immunogenic, as confirmed in a mouse model. Consequently, the present study puts forward a new strategy for the efficient and soluble expression of untagged immunogenic EbolaGP. The intein-based protein fusion approach may be of importance for the large-scale production of Ebola virus subunit vaccine. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. A Versatile System for High-Throughput In Situ X-ray Screening and Data Collection of Soluble and Membrane-Protein Crystals

    Energy Technology Data Exchange (ETDEWEB)

    Broecker, Jana; Klingel, Viviane; Ou, Wei-Lin; Balo, Aidin R.; Kissick, David J.; Ogata, Craig M.; Kuo, Anling; Ernst, Oliver P.

    2016-10-12

    In recent years, in situ data collection has been a major focus of progress in protein crystallography. Here, we introduce the Mylar in situ method using Mylar-based sandwich plates that are inexpensive, easy to make and handle, and show significantly less background scattering than other setups. A variety of cognate holders for patches of Mylar in situ sandwich films corresponding to one or more wells makes the method robust and versatile, allows for storage and shipping of entire wells, and enables automated crystal imaging, screening, and goniometerbased X-ray diffraction data-collection at room temperature and under cryogenic conditions for soluble and membrane-protein crystals grown in or transferred to these plates. We validated the Mylar in situ method using crystals of the water-soluble proteins hen egg-white lysozyme and sperm whale myoglobin as well as the 7-transmembrane protein bacteriorhodopsin from Haloquadratum walsbyi. In conjunction with current developments at synchrotrons, this approach promises high-resolution structural studies of membrane proteins to become faster and more routine.

  1. Protein fiber linear dichroism for structure determination and kinetics in a low-volume, low-wavelength couette flow cell.

    Science.gov (United States)

    Dafforn, Timothy R; Rajendra, Jacindra; Halsall, David J; Serpell, Louise C; Rodger, Alison

    2004-01-01

    High-resolution structure determination of soluble globular proteins relies heavily on x-ray crystallography techniques. Such an approach is often ineffective for investigations into the structure of fibrous proteins as these proteins generally do not crystallize. Thus investigations into fibrous protein structure have relied on less direct methods such as x-ray fiber diffraction and circular dichroism. Ultraviolet linear dichroism has the potential to provide additional information on the structure of such biomolecular systems. However, existing systems are not optimized for the requirements of fibrous proteins. We have designed and built a low-volume (200 microL), low-wavelength (down to 180 nm), low-pathlength (100 microm), high-alignment flow-alignment system (couette) to perform ultraviolet linear dichroism studies on the fibers formed by a range of biomolecules. The apparatus has been tested using a number of proteins for which longer wavelength linear dichroism spectra had already been measured. The new couette cell has also been used to obtain data on two medically important protein fibers, the all-beta-sheet amyloid fibers of the Alzheimer's derived protein Abeta and the long-chain assemblies of alpha1-antitrypsin polymers.

  2. Enhancing production and cytotoxic activity of polymeric soluble FasL-based chimeric proteins by concomitant expression of soluble FasL.

    Directory of Open Access Journals (Sweden)

    Aurore Morello

    Full Text Available Membrane FasL is the natural trigger of Fas-mediated apoptosis. A soluble homotrimeric counterpart (sFasL also exists which is very weakly active, and needs oligomerization beyond its trimeric state to induce apoptosis. We recently generated a soluble FasL chimera by fusing the immunoglobulin-like domain of the leukemia inhibitory factor receptor gp190 to the extracellular region of human FasL, which enabled spontaneous dodecameric homotypic polymerization of FasL. This polymeric soluble human FasL (pFasL displayed anti-tumoral activity in vitro and in vivo without systemic cytotoxicity in mouse. In the present work, we focused on the improvement of pFasL, with two complementary objectives. First, we developed more complex pFasL-based chimeras that contained a cell-targeting module. Secondly, we attempted to improve the production and/or the specific activity of pFasL and of the cell-targeting chimeras. We designed two chimeras by fusing to pFasL the extracellular portions of the HLA-A2 molecule or of a human gamma-delta TCR, and analyzed the consequences of co-expressing these molecules or pFasL together with sFasL on their heterotopic cell production. This strategy significantly enhanced the production of pFasL and of the two chimeras, as well as the cytotoxic activity of the two chimeras but not of pFasL. These results provide the proof of concept for an optimization of FasL-based chimeric proteins for a therapeutic use.

  3. Plasma soluble prion protein, a potential biomarker for sport-related concussions: a pilot study.

    Directory of Open Access Journals (Sweden)

    Nam Pham

    Full Text Available Sport-related mild traumatic brain injury (mTBI or concussion is a significant health concern to athletes with potential long-term consequences. The diagnosis of sport concussion and return to sport decision making is one of the greatest challenges facing health care clinicians working in sports. Blood biomarkers have recently demonstrated their potential in assisting the detection of brain injury particularly, in those cases with no obvious physical injury. We have recently discovered plasma soluble cellular prion protein (PrP(C as a potential reliable biomarker for blast induced TBI (bTBI in a rodent animal model. In order to explore the application of this novel TBI biomarker to sport-related concussion, we conducted a pilot study at the University of Saskatchewan (U of S by recruiting athlete and non-athlete 18 to 30 year-old students. Using a modified quantitative ELISA method, we first established normal values for the plasma soluble PrP(C in male and female students. The measured plasma soluble PrP(C in confirmed concussion cases demonstrated a significant elevation of this analyte in post-concussion samples. Data collected from our pilot study indicates that the plasma soluble PrP(C is a potential biomarker for sport-related concussion, which may be further developed into a clinical diagnostic tool to assist clinicians in the assessment of sport concussion and return-to-play decision making.

  4. Globular and disordered – the non-identical twins in protein-protein interactions

    Directory of Open Access Journals (Sweden)

    Kaare eTeilum

    2015-07-01

    Full Text Available In biology proteins from different structural classes interact across and within classes in ways that are optimized to achieve balanced functional outputs. The interactions between intrinsically disordered proteins (IDPs and other proteins rely on changes in flexibility and this is seen as a strong determinant for their function. This has fostered the notion that IDP’s bind with low affinity but high specificity. Here we have analyzed available detailed thermodynamic data for protein-protein interactions to put to the test if the thermodynamic profiles of IDP interactions differ from those of other protein-protein interactions. We find that ordered proteins and the disordered ones act as non identical twins operating by similar principles but where the disordered proteins complexes are on average less stable by 2.5 kcal mol-1.

  5. Globular adiponectin induces a pro-inflammatory response in human astrocytic cells

    International Nuclear Information System (INIS)

    Wan, Zhongxiao; Mah, Dorrian; Simtchouk, Svetlana; Klegeris, Andis; Little, Jonathan P.

    2014-01-01

    Highlights: • Adiponectin receptors are expressed in human astrocytes. • Globular adiponectin induces secretion of IL-6 and MCP-1 from cultured astrocytes. • Adiponectin may play a pro-inflammatory role in astrocytes. - Abstract: Neuroinflammation, mediated in part by activated brain astrocytes, plays a critical role in the development of neurodegenerative disorders, including Alzheimer’s disease (AD). Adiponectin is the most abundant adipokine secreted from adipose tissue and has been reported to exert both anti- and pro-inflammatory effects in peripheral tissues; however, the effects of adiponectin on astrocytes remain unknown. Shifts in peripheral concentrations of adipokines, including adiponectin, could contribute to the observed link between midlife adiposity and increased AD risk. The aim of the present study was to characterize the effects of globular adiponectin (gAd) on pro-inflammatory cytokine mRNA expression and secretion in human U373 MG astrocytic cells and to explore the potential involvement of nuclear factor (NF)-κB, p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK)1/2, c-Jun N-terminal kinase (JNK) and phosphatidylinositide 3-kinases (PI3 K) signaling pathways in these processes. We demonstrated expression of adiponectin receptor 1 (adipoR1) and adipoR2 in U373 MG cells and primary human astrocytes. gAd induced secretion of interleukin (IL)-6 and monocyte chemoattractant protein (MCP)-1, and gene expression of IL-6, MCP-1, IL-1β and IL-8 in U373 MG cells. Using specific inhibitors, we found that NF-κB, p38MAPK and ERK1/2 pathways are involved in gAd-induced induction of cytokines with ERK1/2 contributing the most. These findings provide evidence that gAd may induce a pro-inflammatory phenotype in human astrocytes

  6. Globular adiponectin induces a pro-inflammatory response in human astrocytic cells

    Energy Technology Data Exchange (ETDEWEB)

    Wan, Zhongxiao; Mah, Dorrian; Simtchouk, Svetlana [School of Health and Exercise Sciences, University of British Columbia Okanagan, Kelowna, BC (Canada); Klegeris, Andis [Department of Biology, University of British Columbia Okanagan, Kelowna, BC (Canada); Little, Jonathan P., E-mail: jonathan.little@ubc.ca [School of Health and Exercise Sciences, University of British Columbia Okanagan, Kelowna, BC (Canada)

    2014-03-28

    Highlights: • Adiponectin receptors are expressed in human astrocytes. • Globular adiponectin induces secretion of IL-6 and MCP-1 from cultured astrocytes. • Adiponectin may play a pro-inflammatory role in astrocytes. - Abstract: Neuroinflammation, mediated in part by activated brain astrocytes, plays a critical role in the development of neurodegenerative disorders, including Alzheimer’s disease (AD). Adiponectin is the most abundant adipokine secreted from adipose tissue and has been reported to exert both anti- and pro-inflammatory effects in peripheral tissues; however, the effects of adiponectin on astrocytes remain unknown. Shifts in peripheral concentrations of adipokines, including adiponectin, could contribute to the observed link between midlife adiposity and increased AD risk. The aim of the present study was to characterize the effects of globular adiponectin (gAd) on pro-inflammatory cytokine mRNA expression and secretion in human U373 MG astrocytic cells and to explore the potential involvement of nuclear factor (NF)-κB, p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK)1/2, c-Jun N-terminal kinase (JNK) and phosphatidylinositide 3-kinases (PI3 K) signaling pathways in these processes. We demonstrated expression of adiponectin receptor 1 (adipoR1) and adipoR2 in U373 MG cells and primary human astrocytes. gAd induced secretion of interleukin (IL)-6 and monocyte chemoattractant protein (MCP)-1, and gene expression of IL-6, MCP-1, IL-1β and IL-8 in U373 MG cells. Using specific inhibitors, we found that NF-κB, p38MAPK and ERK1/2 pathways are involved in gAd-induced induction of cytokines with ERK1/2 contributing the most. These findings provide evidence that gAd may induce a pro-inflammatory phenotype in human astrocytes.

  7. Toxoplasma gondii: Biochemical and biophysical characterization of recombinant soluble dense granule proteins GRA2 and GRA6

    International Nuclear Information System (INIS)

    Bittame, Amina; Effantin, Grégory; Pètre, Graciane; Ruffiot, Pauline; Travier, Laetitia; Schoehn, Guy; Weissenhorn, Winfried; Cesbron-Delauw, Marie-France; Gagnon, Jean; Mercier, Corinne

    2015-01-01

    The most prominent structural feature of the parasitophorous vacuole (PV) in which the intracellular parasite Toxoplasma gondii proliferates is a membranous nanotubular network (MNN), which interconnects the parasites and the PV membrane. The MNN function remains unclear. The GRA2 and GRA6 proteins secreted from the parasite dense granules into the PV have been implicated in the MNN biogenesis. Amphipathic alpha-helices (AAHs) predicted in GRA2 and an alpha-helical hydrophobic domain predicted in GRA6 have been proposed to be responsible for their membrane association, thereby potentially molding the MMN in its structure. Here we report an analysis of the recombinant proteins (expressed in detergent-free conditions) by circular dichroism, which showed that full length GRA2 displays an alpha-helical secondary structure while recombinant GRA6 and GRA2 truncated of its AAHs are mainly random coiled. Dynamic light scattering and transmission electron microscopy showed that recombinant GRA6 and truncated GRA2 constitute a homogenous population of small particles (6–8 nm in diameter) while recombinant GRA2 corresponds to 2 populations of particles (∼8–15 nm and up to 40 nm in diameter, respectively). The unusual properties of GRA2 due to its AAHs are discussed. - Highlights: • Toxoplasma gondii: soluble GRA2 forms 2 populations of particles. • T. gondii: the dense granule protein GRA2 folds intrinsically as an alpha-helix. • T. gondii: monomeric soluble GRA6 forms particles of 6–8 nm in diameter. • T. gondii: monomeric soluble GRA6 is random coiled. • Unusual biophysical properties of the dense granule protein GRA2 from T. gondii

  8. Toxoplasma gondii: Biochemical and biophysical characterization of recombinant soluble dense granule proteins GRA2 and GRA6

    Energy Technology Data Exchange (ETDEWEB)

    Bittame, Amina [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France); Effantin, Grégory [Université Grenoble Alpes, Institut de Biologie Structurale (IBS), 38044 Grenoble (France); CNRS, IBS, 38044 Grenoble (France); CEA, IBS, 38044 Grenoble (France); Unit for Virus Host-Cell Interactions (UVHCI), UMI 3265 (UJF-EMBL-CNRS), 38027 Grenoble (France); Pètre, Graciane; Ruffiot, Pauline; Travier, Laetitia [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France); Schoehn, Guy; Weissenhorn, Winfried [Université Grenoble Alpes, Institut de Biologie Structurale (IBS), 38044 Grenoble (France); CNRS, IBS, 38044 Grenoble (France); CEA, IBS, 38044 Grenoble (France); Unit for Virus Host-Cell Interactions (UVHCI), UMI 3265 (UJF-EMBL-CNRS), 38027 Grenoble (France); Cesbron-Delauw, Marie-France; Gagnon, Jean [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France); Mercier, Corinne, E-mail: corinne.mercier@ujf-grenoble.fr [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France)

    2015-03-27

    The most prominent structural feature of the parasitophorous vacuole (PV) in which the intracellular parasite Toxoplasma gondii proliferates is a membranous nanotubular network (MNN), which interconnects the parasites and the PV membrane. The MNN function remains unclear. The GRA2 and GRA6 proteins secreted from the parasite dense granules into the PV have been implicated in the MNN biogenesis. Amphipathic alpha-helices (AAHs) predicted in GRA2 and an alpha-helical hydrophobic domain predicted in GRA6 have been proposed to be responsible for their membrane association, thereby potentially molding the MMN in its structure. Here we report an analysis of the recombinant proteins (expressed in detergent-free conditions) by circular dichroism, which showed that full length GRA2 displays an alpha-helical secondary structure while recombinant GRA6 and GRA2 truncated of its AAHs are mainly random coiled. Dynamic light scattering and transmission electron microscopy showed that recombinant GRA6 and truncated GRA2 constitute a homogenous population of small particles (6–8 nm in diameter) while recombinant GRA2 corresponds to 2 populations of particles (∼8–15 nm and up to 40 nm in diameter, respectively). The unusual properties of GRA2 due to its AAHs are discussed. - Highlights: • Toxoplasma gondii: soluble GRA2 forms 2 populations of particles. • T. gondii: the dense granule protein GRA2 folds intrinsically as an alpha-helix. • T. gondii: monomeric soluble GRA6 forms particles of 6–8 nm in diameter. • T. gondii: monomeric soluble GRA6 is random coiled. • Unusual biophysical properties of the dense granule protein GRA2 from T. gondii.

  9. An observational study of disk-population globular clusters

    International Nuclear Information System (INIS)

    Armandroff, T.E.

    1988-01-01

    Integrated-light spectroscopy was obtained for twenty-seven globular clusters at the Ca II infrared triplet. Line strengths and radial velocities were measured from the spectra. For the well-studied clusters in the sample, the strength of the CA II lines is very well correlated with previous metallicity estimates obtained using a variety of techniques. The greatly reduced effect of interstellar extinction at these wavelengths compared to the blue region of the spectrum has permitted observations of some of the most heavily reddened clusters in the Galaxy. For several such clusters, the Ca II triplet metallicities are in poor agreement with metallicity estimates from infrared photometry by Malkan. Color-magnitude diagrams were constructed for six previously unstudied metal-rich globular clusters and for the well-studied cluster 47 Tuc. The V magnitudes of the horizontal branch stars in the six clusters are in poor agreement with previous estimates based on secondary methods. The horizontal branch morphologies and reddenings of the program clusters were also determined. Using the improved set of metallicities, radial velocities, and distance moduli, the spatial distribution, kinematics, and metallicity distribution of the Galactic globulars were analyzed. The revised data supports Zinn's conclusion that the metal-rich clusters form a highly flattened, rapidly rotating disk system, while the metal-poor clusters make up the familiar, spherically distributed, slowly rotating halo population. The scale height, metallicity distribution, and kinematics of the metal-rich globulars are in good agreement with those of the stellar thick disk. Luminosity functions were constructed, and no significant difference is found between disk and halo samples. Metallicity gradients seem to be present in the disk cluster system. The implications of these results for the formation and evol

  10. Evaluation of a competitive enzyme-linked immunosorbent assay for measurements of soluble HLA-G protein.

    Science.gov (United States)

    Rasmussen, M; Dahl, M; Buus, S; Djurisic, S; Ohlsson, J; Hviid, T V F

    2014-08-01

    The human leukocyte antigen (HLA) class Ib molecule, HLA-G, has gained increased attention because of its assumed important role in immune regulation. The HLA-G protein exists in several soluble isoforms. Most important are the actively secreted HLA-G5 full-length isoform generated by alternative splicing retaining intron 4 with a premature stop codon, and the cleavage of full-length membrane-bound HLA-G1 from the cell surface, so-called soluble HLA-G1 (sHLA-G1). A specific and sensitive immunoassay for measurements of soluble HLA-G is mandatory for conceivable routine testing and research projects. We report a novel method, a competitive immunoassay, for measuring HLA-G5/sHLA-G1 in biological fluids. The sHLA-G immunoassay is based upon a competitive enzyme-linked immunosorbent assay (ELISA) principle. It includes a recombinant sHLA-G1 protein in complex with β2-microglobulin and a peptide as a standard, biotinylated recombinant sHLA-G1 as an indicator, and the MEM-G/9 anti-HLA-G monoclonal antibody (mAb) as the capture antibody. The specificity and sensitivity of the assay were evaluated. Testing with different recombinant HLA class I proteins and different anti-HLA class I mAbs showed that the sHLA-G immunoassay was highly specific. Optimal combinations of competitor sHLA-G1 and capture mAb concentrations were determined. Two versions of the assay were tested. One with a relatively wide dynamic range from 3.1 to 100.0 ng/ml, and another more sensitive version ranging from 1.6 to 12.5 ng/ml. An intra-assay coefficient of variation (CV) of 15.5% at 88 ng/ml and an inter-assay CV of 23.1% at 39 ng/ml were determined. An assay based on the competitive sHLA-G ELISA may be important for measurements of sHLA-G proteins in several conditions: assisted reproduction, organ transplantation, cancer, and certain pregnancy complications, both in research studies and possibly in the future also for clinical routine use. © 2014 John Wiley & Sons A/S. Published by John Wiley

  11. Rates of collapse and evaporation of globular clusters

    Science.gov (United States)

    Hut, Piet; Djorgovski, S.

    1992-01-01

    Observational estimates of the dynamical relaxation times of Galactic globular clusters are used here to estimate the present rate at which core collapse and evaporation are occurring in them. A core collapse rate of 2 +/- 1 per Gyr is found, which for a Galactic age of about 12 Gyr agrees well with the fact that 27 clusters have surface brightness profiles with the morphology expected for the postcollapse phase. A destruction and evaporation rate of 5 +/- 3 per Gyr is found, suggesting that a significant fraction of the Galaxy's original complement of globular clusters have perished through the combined effects of mechanisms such as relaxation-driven evaporation and shocking due to interaction with the Galactic disk and bulge.

  12. Systematic discovery of new recognition peptides mediating protein interaction networks

    DEFF Research Database (Denmark)

    Neduva, Victor; Linding, Rune; Su-Angrand, Isabelle

    2005-01-01

    Many aspects of cell signalling, trafficking, and targeting are governed by interactions between globular protein domains and short peptide segments. These domains often bind multiple peptides that share a common sequence pattern, or "linear motif" (e.g., SH3 binding to PxxP). Many domains...... by interactions between globular protein domains and short peptide segments. These domains often bind multiple peptides that share a common sequence pattern, or "linear motif" (e.g., SH3 binding to PxxP). Many domains are known, though comparatively few linear motifs have been discovered. Their short length...

  13. Delayed Toxicity Associated with Soluble Anthrax Toxin Receptor Decoy-Ig Fusion Protein Treatment

    Science.gov (United States)

    Cote, Christopher; Welkos, Susan; Manchester, Marianne; Young, John A. T.

    2012-01-01

    Soluble receptor decoy inhibitors, including receptor-immunogloubulin (Ig) fusion proteins, have shown promise as candidate anthrax toxin therapeutics. These agents act by binding to the receptor-interaction site on the protective antigen (PA) toxin subunit, thereby blocking toxin binding to cell surface receptors. Here we have made the surprising observation that co-administration of receptor decoy-Ig fusion proteins significantly delayed, but did not protect, rats challenged with anthrax lethal toxin. The delayed toxicity was associated with the in vivo assembly of a long-lived complex comprised of anthrax lethal toxin and the receptor decoy-Ig inhibitor. Intoxication in this system presumably results from the slow dissociation of the toxin complex from the inhibitor following their prolonged circulation. We conclude that while receptor decoy-Ig proteins represent promising candidates for the early treatment of B. anthracis infection, they may not be suitable for therapeutic use at later stages when fatal levels of toxin have already accumulated in the bloodstream. PMID:22511955

  14. Effect of radiation pressure in the cores of globular clusters

    Energy Technology Data Exchange (ETDEWEB)

    Angeletti, L; Capuzzo-Dolcetta, R; Giannone

    1981-10-01

    The possible effects of a presence of a dust cloud in the cores of globular clusters was investigated. Two cluster models were considered together with various models of clouds. The problem of radiation transfer was solved under some simplifying assumptions. Owing to a differential absorption of the star light in the cloud, radiation pressure turned out be inward-directed in some cloud models. This fact may lead to a confinement of some dust in the central regions of globular clusters.

  15. Globular clusters as a source of X-ray emission from the neighbourhood of M87

    International Nuclear Information System (INIS)

    Fabian, A.C.; Pringle, J.E.; Rees, M.J.

    1976-01-01

    It is stated that the X-ray emission from globular clusters may be attributable to accretion on to compact objects, the accreting material being supplied from binary companions, or gas trapped in the potential well of the cluster. Counts of objects in the vicinity of the M87 have revealed that it has an extensive halo of globular clusters, the number of which may exceed 10,000 within a radius of 23 arc min. Most of these clusters may be explicable as a population effect, and the similarity of their optical properties to those of cluster in our own Galaxy suggests that they may also contain X-ray sources. The brighter globular clusters in M87 may, however, be substantially more X-ray luminous, and there may be proportionally more gas available in globular clusters in M87 compared with our Galaxy. The average X-ray luminosity of individual globular clusters may be of the order of 10 38 erg/sec., which raises the possibility that the integrated globular cluster emission may account for a substantial fraction of the X-ray emission observed from the region of M87. In support of this it is noted that the extended X-ray emission from the Virgo cluster is centered on M87, which lies approximately 45 arc min from the cluster centroid, and it is expected that the general X-ray emission from the globular cluster will appear to be smoothly and symmetrically distributed about M87 at moderate spatial resolution. A similar situation may apply to the elliptical galaxy NGC 3311 in Abell 1060 which, as a cluster, has been suggested as the identification for the X-ray source 3 U 1044-40, and it seems possible that that galaxy is surrounded by a similar globular cluster population to that of M87. (U.K.)

  16. Protein- protein interaction detection system using fluorescent protein microdomains

    Science.gov (United States)

    Waldo, Geoffrey S.; Cabantous, Stephanie

    2010-02-23

    The invention provides a protein labeling and interaction detection system based on engineered fragments of fluorescent and chromophoric proteins that require fused interacting polypeptides to drive the association of the fragments, and further are soluble and stable, and do not change the solubility of polypeptides to which they are fused. In one embodiment, a test protein X is fused to a sixteen amino acid fragment of GFP (.beta.-strand 10, amino acids 198-214), engineered to not perturb fusion protein solubility. A second test protein Y is fused to a sixteen amino acid fragment of GFP (.beta.-strand 11, amino acids 215-230), engineered to not perturb fusion protein solubility. When X and Y interact, they bring the GFP strands into proximity, and are detected by complementation with a third GFP fragment consisting of GFP amino acids 1-198 (strands 1-9). When GFP strands 10 and 11 are held together by interaction of protein X and Y, they spontaneous association with GFP strands 1-9, resulting in structural complementation, folding, and concomitant GFP fluorescence.

  17. Identification of Proteins with Potential Osteogenic Activity Present in the Water-Soluble Matrix Proteins from Crassostrea gigas Nacre Using a Proteomic Approach

    Directory of Open Access Journals (Sweden)

    Daniel V. Oliveira

    2012-01-01

    Full Text Available Nacre, when implanted in vivo in bones of dogs, sheep, mice, and humans, induces a biological response that includes integration and osteogenic activity on the host tissue that seems to be activated by a set of proteins present in the nacre water-soluble matrix (WSM. We describe here an experimental approach that can accurately identify the proteins present in the WSM of shell mollusk nacre. Four proteins (three gigasin-2 isoforms and a cystatin A2 were for the first time identified in WSM of Crassostrea gigas nacre using 2DE and LC-MS/MS for protein identification. These proteins are thought to be involved in bone remodeling processes and could be responsible for the biocompatibility shown between bone and nacre grafts. These results represent a contribution to the study of shell biomineralization process and opens new perspectives for the development of new nacre biomaterials for orthopedic applications.

  18. Chemical Abundances of Giants in Globular Clusters

    Science.gov (United States)

    Gratton, Raffaele G.; Bragaglia, Angela; Carretta, Eugenio; D'Orazi, Valentina; Lucatello, Sara

    A large fraction of stars form in clusters. According to a widespread paradigma, stellar clusters are prototypes of single stellar populations. According to this concept, they formed on a very short time scale, and all their stars share the same chemical composition. Recently it has been understood that massive stellar clusters (the globular clusters) rather host various stellar populations, characterized by different chemical composition: these stellar populations have also slightly different ages, stars of the second generations being formed from the ejecta of part of those of an earlier one. Furthermore, it is becoming clear that the efficiency of the process is quite low: many more stars formed within this process than currently present in the clusters. This implies that a significant, perhaps even dominant fraction of the ancient population of galaxies formed within the episodes that lead to formation the globular clusters.

  19. Small surfactant-like peptides can drive soluble proteins into active aggregates

    Directory of Open Access Journals (Sweden)

    Zhou Bihong

    2012-01-01

    Full Text Available Abstract Background Inactive protein inclusion bodies occur commonly in Escherichia coli (E. coli cells expressing heterologous proteins. Previously several independent groups have found that active protein aggregates or pseudo inclusion bodies can be induced by a fusion partner such as a cellulose binding domain from Clostridium cellulovorans (CBDclos when expressed in E. coli. More recently we further showed that a short amphipathic helical octadecapeptide 18A (EWLKAFYEKVLEKLKELF and a short beta structure peptide ELK16 (LELELKLKLELELKLK have a similar property. Results In this work, we explored a third type of peptides, surfactant-like peptides, for performing such a "pulling-down" function. One or more of three such peptides (L6KD, L6K2, DKL6 were fused to the carboxyl termini of model proteins including Aspergillus fumigatus amadoriase II (AMA, all three peptides were used, Bacillus subtilis lipase A (LipA, only L6KD was used, hereinafter the same, Bacillus pumilus xylosidase (XynB, and green fluorescent protein (GFP, and expressed in E. coli. All fusions were found to predominantly accumulate in the insoluble fractions, with specific activities ranging from 25% to 92% of the native counterparts. Transmission electron microscopic (TEM and confocal fluorescence microscopic analyses confirmed the formation of protein aggregates in the cell. Furthermore, binding assays with amyloid-specific dyes (thioflavin T and Cong red to the AMA-L6KD aggregate and the TEM analysis of the aggregate following digestion with protease K suggested that the AMA-L6KD aggregate may contain structures reminiscent of amyloids, including a fibril-like structure core. Conclusions This study shows that the surfactant-like peptides L6KD and it derivatives can act as a pull-down handler for converting soluble proteins into active aggregates, much like 18A and ELK16. These peptide-mediated protein aggregations might have important implications for protein aggregation in

  20. The evoluation of the galactic globular clusters; I Metal abundance calibrations

    International Nuclear Information System (INIS)

    Lee, S.W.; Park, N.K.

    1984-01-01

    Five different calibrations of metal abundances of globular clusters are examined and these are compared with metallicity ranking parameters such as (Sp)sub(c), , Q39 and IR-indices. Except for the calibration *(Fe/H*)sub(H) by the high dispersion echelle analysis, the other calibration scales are correlated with the morphological parameters of red giant branch. In the *(Fe/H*)sub(Hsup(-))scale, the clusters later than approx.F8 have nearly a constant metal abundance, *(Fe/H*)sub(H)approx.-1.05, regardless of morphological characteristics of horizontal branch and red giant branch. By the two fundamental calibration scales of *(Fe/H*)sub(L) (derived by the low dispersion analysis), and *(Fe/H*)sub(delta S) (derived by the spectral analysis of RR Lyrae stars), the globular clusters are divided into the halo clusters with *(Fe/H*)<-1.0 and the disk clusters confined within the galactocentric distance rsub(G)=10 kpc and galactic plane distance absolute z=3 kpc. In this case the abundance gradient is given by d*(Fe/H*)/drsub(G)approx.-0.05kpcsup(-1) and d*(Fe/H*)/d absolute z approx. -0.08 kpcsup(-1) within rsub(G)=20 kpc and absolute z=10 kpc, respectively. According to these characteristics of the spatial distribution of globular clusters, the chemical evolution of the galactic globular clusters can be accounted for by the two-zone (disk-halo) slow collapse model when the *(Fe/H*)sub(Lsup(-)) or *(Fe/H*)sub(DELTA Ssup(-))scale is applied. In the case of *(Fe/H*)sub(Hsup(-))scale, the one-zone fast collapse model is preferred for the evolution of globular clusters. (Author)

  1. A catalogue of masses, structural parameters and velocity dispersion profiles of 112 Milky Way globular clusters

    Science.gov (United States)

    Baumgardt, H.; Hilker, M.

    2018-05-01

    We have determined masses, stellar mass functions and structural parameters of 112 Milky Way globular clusters by fitting a large set of N-body simulations to their velocity dispersion and surface density profiles. The velocity dispersion profiles were calculated based on a combination of more than 15,000 high-precision radial velocities which we derived from archival ESO/VLT and Keck spectra together with ˜20, 000 published radial velocities from the literature. Our fits also include the stellar mass functions of the globular clusters, which are available for 47 clusters in our sample, allowing us to self-consistently take the effects of mass segregation and ongoing cluster dissolution into account. We confirm the strong correlation between the global mass functions of globular clusters and their relaxation times recently found by Sollima & Baumgardt (2017). We also find a correlation of the escape velocity from the centre of a globular cluster and the fraction of first generation stars (FG) in the cluster recently derived for 57 globular clusters by Milone et al. (2017), but no correlation between the FG star fraction and the global mass function of a globular cluster. This could indicate that the ability of a globular cluster to keep the wind ejecta from the polluting star(s) is the crucial parameter determining the presence and fraction of second generation stars and not its later dynamical mass loss.

  2. Binary Black Hole Mergers from Globular Clusters: Implications for Advanced LIGO.

    Science.gov (United States)

    Rodriguez, Carl L; Morscher, Meagan; Pattabiraman, Bharath; Chatterjee, Sourav; Haster, Carl-Johan; Rasio, Frederic A

    2015-07-31

    The predicted rate of binary black hole mergers from galactic fields can vary over several orders of magnitude and is extremely sensitive to the assumptions of stellar evolution. But in dense stellar environments such as globular clusters, binary black holes form by well-understood gravitational interactions. In this Letter, we study the formation of black hole binaries in an extensive collection of realistic globular cluster models. By comparing these models to observed Milky Way and extragalactic globular clusters, we find that the mergers of dynamically formed binaries could be detected at a rate of ∼100 per year, potentially dominating the binary black hole merger rate. We also find that a majority of cluster-formed binaries are more massive than their field-formed counterparts, suggesting that Advanced LIGO could identify certain binaries as originating from dense stellar environments.

  3. A simple model for electrical charge in globular macromolecules and linear polyelectrolytes in solution

    Science.gov (United States)

    Krishnan, M.

    2017-05-01

    We present a model for calculating the net and effective electrical charge of globular macromolecules and linear polyelectrolytes such as proteins and DNA, given the concentration of monovalent salt and pH in solution. The calculation is based on a numerical solution of the non-linear Poisson-Boltzmann equation using a finite element discretized continuum approach. The model simultaneously addresses the phenomena of charge regulation and renormalization, both of which underpin the electrostatics of biomolecules in solution. We show that while charge regulation addresses the true electrical charge of a molecule arising from the acid-base equilibria of its ionizable groups, charge renormalization finds relevance in the context of a molecule's interaction with another charged entity. Writing this electrostatic interaction free energy in terms of a local electrical potential, we obtain an "interaction charge" for the molecule which we demonstrate agrees closely with the "effective charge" discussed in charge renormalization and counterion-condensation theories. The predictions of this model agree well with direct high-precision measurements of effective electrical charge of polyelectrolytes such as nucleic acids and disordered proteins in solution, without tunable parameters. Including the effective interior dielectric constant for compactly folded molecules as a tunable parameter, the model captures measurements of effective charge as well as published trends of pKa shifts in globular proteins. Our results suggest a straightforward general framework to model electrostatics in biomolecules in solution. In offering a platform that directly links theory and experiment, these calculations could foster a systematic understanding of the interrelationship between molecular 3D structure and conformation, electrical charge and electrostatic interactions in solution. The model could find particular relevance in situations where molecular crystal structures are not available or

  4. ORF-selector ESPRIT: a second generation library screen for soluble protein expression employing precise open reading frame selection.

    Science.gov (United States)

    An, Yingfeng; Yumerefendi, Hayretin; Mas, Philippe J; Chesneau, Alban; Hart, Darren J

    2011-08-01

    Here we present ORF-selector ESPRIT, a 9-fold enhanced version of our technology for screening incremental truncation libraries to identify soluble high yielding constructs of challenging proteins. Gene fragments are truncated at both termini to access internal domains and the resulting reading frame problem is addressed by an unbiased, intein-based open reading frame selection yielding only in-frame DNA inserts. This enriched library is then subcloned into a standard high-level expression plasmid where tens of thousands of constructs can be assayed in a two-step process using colony- and liquid-handling robots to isolate rare highly expressing clones useful for production of multi milligram quantities of purifiable proteins. The p85α protein was used to benchmark the system resulting in isolation of all known domains, either alone or in tandem. The human kinase IKK1 was then screened resulting in purification of a predicted internal domain. This strategy provides an integrated, facile route to produce soluble proteins from challenging and poorly understood target genes at quantities compatible with structural biology, screening applications and immunisation studies. The high genetic diversity that can be sampled opens the way to study more diverse systems including multisubunit complexes. Copyright © 2011 Elsevier Inc. All rights reserved.

  5. On tidal radius determination for a globular cluster

    International Nuclear Information System (INIS)

    Ninkovic, S.

    1985-01-01

    A tidal radius determination for a globular cluster based on its density minimum, which is caused by the galactic tidal forces and derivable from a model of the Galaxy, is proposed. Results obtained on the basis of the Schmidt model for two clusters are in a satisfactory agreement with those obtained earlier by means of other methods. A mass determination for the clusters through the tidal radius, when the latter one is identified with the cluster perigalactic distance, yields unusually large mass values. Probably, the tidal radius should be identified with the instantaneous galactocentric distance. Use of models more recent than the Schmidt one indicates that a globular cluster may contain a significant portion of an invisible interstellar matter. (author)

  6. Globular Cluster Candidates for Hosting a Central Black Hole

    Science.gov (United States)

    Noyola, Eva

    2009-07-01

    We are continuing our study of the dynamical properties of globular clusters and we propose to obtain surface brightness profiles for high concentration clusters. Our results to date show that the distribution of central surface brightness slopes do not conform to standard models. This has important implications for how they form and evolve, and suggest the possible presence of central intermediate-mass black holes. From our previous archival proposals {AR-9542 and AR-10315}, we find that many high concentration globular clusters do not have flat cores or steep central cusps, instead they show weak cusps. Numerical simulations suggest that clusters with weak cusps may harbor intermediate-mass black holes and we have one confirmation of this connection with omega Centauri. This cluster shows a shallow cusp in its surface brightness profile, while kinematical measurements suggest the presence of a black hole in its center. Our goal is to extend these studies to a sample containing 85% of the Galactic globular clusters with concentrations higher than 1.7 and look for objects departing from isothermal behavior. The ACS globular cluster survey {GO-10775} provides enough objects to have an excellent coverage of a wide range of galactic clusters, but it contains only a couple of the ones with high concentration. The proposed sample consists of clusters whose light profile can only be adequately measured from space-based imaging. This would take us close to completeness for the high concentration cases and therefore provide a more complete list of candidates for containing a central black hole. The dataset will also be combined with our existing kinematic measurements and enhanced with future kinematic studies to perform detailed dynamical modeling.

  7. HUBBLE PINPOINTS WHITE DWARFS IN GLOBULAR CLUSTER

    Science.gov (United States)

    2002-01-01

    Peering deep inside a cluster of several hundred thousand stars, NASA's Hubble Space Telescope uncovered the oldest burned-out stars in our Milky Way Galaxy. Located in the globular cluster M4, these small, dying stars - called white dwarfs - are giving astronomers a fresh reading on one of the biggest questions in astronomy: How old is the universe? The ancient white dwarfs in M4 are about 12 to 13 billion years old. After accounting for the time it took the cluster to form after the big bang, astronomers found that the age of the white dwarfs agrees with previous estimates for the universe's age. In the top panel, a ground-based observatory snapped a panoramic view of the entire cluster, which contains several hundred thousand stars within a volume of 10 to 30 light-years across. The Kitt Peak National Observatory's 0.9-meter telescope took this picture in March 1995. The box at left indicates the region observed by the Hubble telescope. The Hubble telescope studied a small region of the cluster. A section of that region is seen in the picture at bottom left. A sampling of an even smaller region is shown at bottom right. This region is only about one light-year across. In this smaller region, Hubble pinpointed a number of faint white dwarfs. The blue circles pinpoint the dwarfs. It took nearly eight days of exposure time over a 67-day period to find these extremely faint stars. Globular clusters are among the oldest clusters of stars in the universe. The faintest and coolest white dwarfs within globular clusters can yield a globular cluster's age. Earlier Hubble observations showed that the first stars formed less than 1 billion years after the universe's birth in the big bang. So, finding the oldest stars puts astronomers within arm's reach of the universe's age. M4 is 7,000 light-years away in the constellation Scorpius. Hubble's Wide Field and Planetary Camera 2 made the observations from January through April 2001. These optical observations were combined to

  8. Identification of Hard X-ray Sources in Galactic Globular Clusters: Simbol-X Simulations

    Science.gov (United States)

    Servillat, M.

    2009-05-01

    Globular clusters harbour an excess of X-ray sources compared to the number of X-ray sources in the Galactic plane. It has been proposed that many of these X-ray sources are cataclysmic variables that have an intermediate magnetic field, i.e. intermediate polars, which remains to be confirmed and understood. We present here several methods to identify intermediate polars in globular clusters from multiwavelength analysis. First, we report on XMM-Newton, Chandra and HST observations of the very dense Galactic globular cluster NGC 2808. By comparing UV and X-ray properties of the cataclysmic variable candidates, the fraction of intermediate polars in this cluster can be estimated. We also present the optical spectra of two cataclysmic variables in the globular cluster M 22. The HeII (4868 Å) emission line in these spectra could be related to the presence of a magnetic field in these objects. Simulations of Simbol-X observations indicate that the angular resolution is sufficient to study X-ray sources in the core of close, less dense globular clusters, such as M 22. The sensitivity of Simbol-X in an extended energy band up to 80 keV will allow us to discriminate between hard X-ray sources (such as magnetic cataclysmic variables) and soft X-ray sources (such as chromospherically active binaries).

  9. Depletion-induced instability in protein-DNA mixtures: Influence of protein charge and size

    NARCIS (Netherlands)

    Vries, de R.J.

    2006-01-01

    While there is abundant experimental and theoretical work on polymer-induced DNA condensation, it is still unclear whether globular proteins can condense linear DNA or not. We develop a simple analytical approximation for the depletion attraction between rodlike segments of semiflexible

  10. Integrated-light spectroscopy of globular clusters at the infrared Ca II lines

    Science.gov (United States)

    Armandroff, Taft E.; Zinn, Robert

    1988-01-01

    Integrated-light spectroscopy has been obtained for 27 globular clusters at the Ca II IR triplet. Line strengths and radial velocities have been measured from the spectra. For the well-studied clusters in the sample, the strength of the Ca II lines is very well correlated with previous metallicity estimates. Thus, the triplet is useful as a metallicity indicator in globular cluster integrated-light spectra. The greatly reduced effect of interstellar extinction at these wavelengths (compared to the blue region of the spectrum) has permitted observations of some of the most heavily reddened clusters in the Galaxy. For several such clusters, the Ca II triplet metallicities are in poor agreement with metallicity estimates from IR photometry by Malkan (1981). The strength of an interstellar band at 8621A has been used to estimate the amount of extinction towards these clusters. Using the new metallicity and radial-velocity data, the metallicity distribution, kinematics, and spatial distribution of the disk globular cluster system have been analyzed. Results very similar to those of Zinn (1985) have been found. The relation of the disk globulars to the stellar thick disk is discussed.

  11. Disrupted globular clusters and the gamma-ray excess in the Galactic Centre

    Science.gov (United States)

    Fragione, Giacomo; Antonini, Fabio; Gnedin, Oleg Y.

    2018-04-01

    The Fermi Large Area Telescope has provided the most detailed view towards the Galactic Centre (GC) in high-energy gamma-rays. Besides the interstellar emission and point source contributions, the data suggest a residual diffuse gamma-ray excess. The similarity of its spatial distribution with the expected profile of dark matter has led to claims that this may be evidence for dark matter particle annihilation. Here, we investigate an alternative explanation that the signal originates from millisecond pulsars (MSPs) formed in dense globular clusters and deposited at the GC as a consequence of cluster inspiral and tidal disruption. We use a semi-analytical model to calculate the formation, migration, and disruption of globular clusters in the Galaxy. Our model reproduces the mass of the nuclear star cluster and the present-day radial and mass distribution of globular clusters. For the first time, we calculate the evolution of MSPs from disrupted globular clusters throughout the age of the Galaxy and consistently include the effect of the MSP spin-down due to magnetic-dipole braking. The final gamma-ray amplitude and spatial distribution are in good agreement with the Fermi observations and provide a natural astrophysical explanation for the GC excess.

  12. Application of far-infrared spectroscopy to the structural identification of protein materials.

    Science.gov (United States)

    Han, Yanchen; Ling, Shengjie; Qi, Zeming; Shao, Zhengzhong; Chen, Xin

    2018-05-03

    Although far-infrared (IR) spectroscopy has been shown to be a powerful tool to determine peptide structure and to detect structural transitions in peptides, it has been overlooked in the characterization of proteins. Herein, we used far-IR spectroscopy to monitor the structure of four abundant non-bioactive proteins, namely, soybean protein isolate (SPI), pea protein isolate (PPI) and two types of silk fibroins (SFs), domestic Bombyx mori and wild Antheraea pernyi. The two globular proteins SPI and PPI result in broad and weak far-IR bands (between 50 and 700 cm-1), in agreement with those of some other bioactive globular proteins previously studied (lysozyme, myoglobin, hemoglobin, etc.) that generally only have random amino acid sequences. Interestingly, the two SFs, which are characterized by a structure composed of highly repetitive motifs, show several sharp far-IR characteristic absorption peaks. Moreover, some of these characteristic peaks (such as the peaks at 260 and 428 cm-1 in B. mori, and the peaks at 245 and 448 cm-1 in A. pernyi) are sensitive to conformational changes; hence, they can be directly used to monitor conformational transitions in SFs. Furthermore, since SF absorption bands clearly differ from those of globular proteins and different SFs even show distinct adsorption bands, far-IR spectroscopy can be applied to distinguish and determine the specific SF component within protein blends.

  13. The ellipticities of a sample of globular clusters in M31

    International Nuclear Information System (INIS)

    Lupton, R.H.

    1989-01-01

    Images for a sample of 18 globular clusters in M31 have been obtained. The mean ellipticity on the sky in the range 7-14 pc (2-4 arcsec) is 0.08 + or - 0.02 and 0.12 + or - 0.01 in the range 14-21 pc (4-6 arcsec), with corresponding true ellipticities of 0.12 and 0.18. The difference between the inner and outer parts is significant at a 99 percent level. The flattening of the inner parts is statistically indistinguishable from that of the Galactic globular clusters, while the outer parts are flatter than the Galactic clusters at a 99.8 percent confidence level. There is a significant anticorrelation of ellipticity with line strength; such a correlation may in retrospect also be seen in the Galactic globular cluster system. For the M31 data, this anticorrelation is stronger in the inner parts of the galaxy. 30 refs

  14. The Frequency of Binary Stars in the Globular Cluster M71

    Science.gov (United States)

    Barden, S. C.; Armandroff, T. E.; Pryor, C. P.

    1994-12-01

    The frequency of binary stars is a fundamental property of a stellar population. A comparison of the frequency of binaries in globular clusters with those in the field halo and disk populations tests the similarity of star formation in those environments. Binary stars in globular clusters also act as an energy source which ``heats" the cluster through super-elastic encounters with other stars and binaries. Such encounters can not only profoundly affect the dynamical evolution of the cluster, they can disrupt the widely separated binaries and catalyze the formation of exotic objects such as blue stragglers, x-ray binaries, and milli-second pulsars. We have used the KPNO 4-m and the multi-fiber instruments Nessie and Hydra to measure radial velocities at 4 epochs over two years for a sample of 126 stars in the globular cluster M71. Velocity errors are under 1 km s(-1) for the brighter stars and under 2 km s(-1) for the majority of our data set. These velocities will be valuable for studying the kinematics of M71, but here we focus on searching for binaries. The faintest stars are at V=17, or just above the main sequence turnoff. Our sample is thus deeper than any published globular cluster binary search utilizing spectroscopic techniques. By observing smaller stars, we double the number of decades of binary periods sampled compared to previous studies and come within a factor of 4 of the shortest possible periods for turnoff stars. This wider period window has produced the largest known sample of binaries in a globular cluster. Four stars show velocity ranges larger than 20 km s(-1) , nine have ranges larger than 10 km s(-1) , and others are clearly variable. We will compare the radial distribution of these stars to that predicted by theory and derive the main-sequence binary fraction.

  15. Recombinant Protein Truncation Strategy for Inducing Bactericidal Antibodies to the Macrophage Infectivity Potentiator Protein of Neisseria meningitidis and Circumventing Potential Cross-Reactivity with Human FK506-Binding Proteins

    Science.gov (United States)

    Bielecka, Magdalena K.; Devos, Nathalie; Gilbert, Mélanie; Hung, Miao-Chiu; Weynants, Vincent; Heckels, John E.

    2014-01-01

    A recombinant macrophage infectivity potentiator (rMIP) protein of Neisseria meningitidis induces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity to human FK506-binding proteins (FKBPs) in residues 166 to 252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognize human proteins, we immunized mice with recombinant truncated type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1 to 22) and contained the His purification tag at either the N or C terminus (C-term). The immunogenicity of truncated rMIP proteins was compared to that of full (i.e., full-length) rMIP proteins (containing the globular domain) with either an N- or C-terminal His tag and with or without the LP sequence. By comparing the functional murine antibody responses to these various constructs, we determined that C-term His truncated rMIP (−LP) delivered in liposomes induced high levels of antibodies that bound to the surface of wild-type but not Δmip mutant meningococci and showed bactericidal activity against homologous type I MIP (median titers of 128 to 256) and heterologous type II and III (median titers of 256 to 512) strains, thereby providing at least 82% serogroup B strain coverage. In contrast, in constructs lacking the LP, placement of the His tag at the N terminus appeared to abrogate bactericidal activity. The strategy used in this study would obviate any potential concerns regarding the use of MIP antigens for inclusion in bacterial vaccines. PMID:25452551

  16. Recombinant protein truncation strategy for inducing bactericidal antibodies to the macrophage infectivity potentiator protein of Neisseria meningitidis and circumventing potential cross-reactivity with human FK506-binding proteins.

    Science.gov (United States)

    Bielecka, Magdalena K; Devos, Nathalie; Gilbert, Mélanie; Hung, Miao-Chiu; Weynants, Vincent; Heckels, John E; Christodoulides, Myron

    2015-02-01

    A recombinant macrophage infectivity potentiator (rMIP) protein of Neisseria meningitidis induces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity to human FK506-binding proteins (FKBPs) in residues 166 to 252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognize human proteins, we immunized mice with recombinant truncated type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1 to 22) and contained the His purification tag at either the N or C terminus (C-term). The immunogenicity of truncated rMIP proteins was compared to that of full (i.e., full-length) rMIP proteins (containing the globular domain) with either an N- or C-terminal His tag and with or without the LP sequence. By comparing the functional murine antibody responses to these various constructs, we determined that C-term His truncated rMIP (-LP) delivered in liposomes induced high levels of antibodies that bound to the surface of wild-type but not Δmip mutant meningococci and showed bactericidal activity against homologous type I MIP (median titers of 128 to 256) and heterologous type II and III (median titers of 256 to 512) strains, thereby providing at least 82% serogroup B strain coverage. In contrast, in constructs lacking the LP, placement of the His tag at the N terminus appeared to abrogate bactericidal activity. The strategy used in this study would obviate any potential concerns regarding the use of MIP antigens for inclusion in bacterial vaccines. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. Interdependence of the rad50 hook and globular domain functions.

    Science.gov (United States)

    Hohl, Marcel; Kochańczyk, Tomasz; Tous, Cristina; Aguilera, Andrés; Krężel, Artur; Petrini, John H J

    2015-02-05

    Rad50 contains a conserved Zn(2+) coordination domain (the Rad50 hook) that functions as a homodimerization interface. Hook ablation phenocopies Rad50 deficiency in all respects. Here, we focused on rad50 mutations flanking the Zn(2+)-coordinating hook cysteines. These mutants impaired hook-mediated dimerization, but recombination between sister chromatids was largely unaffected. This may reflect that cohesin-mediated sister chromatid interactions are sufficient for double-strand break repair. However, Mre11 complex functions specified by the globular domain, including Tel1 (ATM) activation, nonhomologous end joining, and DNA double-strand break end resection were affected, suggesting that dimerization exerts a broad influence on Mre11 complex function. These phenotypes were suppressed by mutations within the coiled-coil and globular ATPase domains, suggesting a model in which conformational changes in the hook and globular domains are transmitted via the extended coils of Rad50. We propose that transmission of spatial information in this manner underlies the regulation of Mre11 complex functions. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. The Age of Globular Clusters in Light of Hipparcos: Resolving the Age Problem?

    Science.gov (United States)

    Chaboyer, Brian; Demarque, P.; Kernan, Peter J.; Krauss, Lawrence M.

    1998-02-01

    We review five independent techniques that are used to set the distance scale to globular clusters, including subdwarf main-sequence fitting utilizing the recent Hipparcos parallax catalog. These data together all indicate that globular clusters are farther away than previously believed, implying a reduction in age estimates. We now adopt a best-fit value Mv (RR Lyrae stars) = 0.39 +/- 0.08 (statistical) at [Fe/H] = -1.9 with an additional uniform systematic uncertainty of +0.13-0.18. This new distance scale estimate is combined with a detailed numerical Monte Carlo study (previously reported by Chaboyer et al.) designed to assess the uncertainty associated with the theoretical age-turnoff luminosity relationship in order to estimate both the absolute age and uncertainty in age of the oldest globular clusters. Our best estimate for the mean age of the oldest globular clusters is now 11.5 +/- 1.3 Gyr, with a one-sided 95% confidence level lower limit of 9.5 Gyr. This represents a systematic shift of over 2 σ compared to our earlier estimate, owing completely to the new distance scale--a shift which we emphasize results not only from the Hipparcos data. This now provides a lower limit on the age of the universe that is consistent with either an open universe or with a flat matter-dominated universe (the latter requiring H0 explicitly quantifies how remaining uncertainties in the distance scale and stellar evolution models translate into uncertainties in the derived globular cluster ages. Simple formulae are provided that can be used to update our age estimate as improved determinations for various quantities become available. Formulae are also provided that can be used to derive the age and its uncertainty for a globular cluster, given the absolute magnitude of the turnoff or the point on the subgiant branch 0.05 mag redder than the turnoff.

  19. A sparse autoencoder-based deep neural network for protein solvent accessibility and contact number prediction.

    Science.gov (United States)

    Deng, Lei; Fan, Chao; Zeng, Zhiwen

    2017-12-28

    Direct prediction of the three-dimensional (3D) structures of proteins from one-dimensional (1D) sequences is a challenging problem. Significant structural characteristics such as solvent accessibility and contact number are essential for deriving restrains in modeling protein folding and protein 3D structure. Thus, accurately predicting these features is a critical step for 3D protein structure building. In this study, we present DeepSacon, a computational method that can effectively predict protein solvent accessibility and contact number by using a deep neural network, which is built based on stacked autoencoder and a dropout method. The results demonstrate that our proposed DeepSacon achieves a significant improvement in the prediction quality compared with the state-of-the-art methods. We obtain 0.70 three-state accuracy for solvent accessibility, 0.33 15-state accuracy and 0.74 Pearson Correlation Coefficient (PCC) for the contact number on the 5729 monomeric soluble globular protein dataset. We also evaluate the performance on the CASP11 benchmark dataset, DeepSacon achieves 0.68 three-state accuracy and 0.69 PCC for solvent accessibility and contact number, respectively. We have shown that DeepSacon can reliably predict solvent accessibility and contact number with stacked sparse autoencoder and a dropout approach.

  20. Measuring age differences among globular clusters having similar metallicities - A new method and first results

    International Nuclear Information System (INIS)

    Vandenberg, D.A.; Bolte, M.; Stetson, P.B.

    1990-01-01

    A color-difference technique for estimating the relative ages of globular clusters with similar chemical compositions on the basis of their CM diagrams is described and demonstrated. The theoretical basis and implementation of the procedure are explained, and results for groups of globular clusters with m/H = about -2, -1.6, and -1.3, and for two special cases (Palomar 12 and NGC 5139) are presented in extensive tables and graphs and discussed in detail. It is found that the more metal-deficient globular clusters are nearly coeval (differences less than 0.5 Gyr), whereas the most metal-rich globular clusters exhibit significant age differences (about 2 Gyr). This result is shown to contradict Galactic evolution models postulating halo collapse in less than a few times 100 Myr. 77 refs

  1. Recombinant Protein Truncation Strategy for Inducing Bactericidal Antibodies to the Macrophage Infectivity Potentiator Protein of Neisseria meningitidis and Circumventing Potential Cross-Reactivity with Human FK506-Binding Proteins

    OpenAIRE

    Bielecka, Magdalena K.; Devos, Nathalie; Gilbert, Mélanie; Hung, Miao-Chiu; Weynants, Vincent; Heckels, John E.; Christodoulides, Myron

    2014-01-01

    A recombinant macrophage infectivity potentiator (rMIP) protein of Neisseria meningitidis induces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity to human FK506-binding proteins (FKBPs) in residues 166 to 252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognize human protein...

  2. Water-Soluble Chlorophyll Protein (WSCP) Stably Binds Two or Four Chlorophylls.

    Science.gov (United States)

    Palm, Daniel M; Agostini, Alessandro; Tenzer, Stefan; Gloeckle, Barbara M; Werwie, Mara; Carbonera, Donatella; Paulsen, Harald

    2017-03-28

    Water-soluble chlorophyll proteins (WSCPs) of class IIa from Brassicaceae form tetrameric complexes containing one chlorophyll (Chl) per apoprotein but no carotenoids. The complexes are remarkably stable toward dissociation and protein denaturation even at 100 °C and extreme pH values, and the Chls are partially protected against photooxidation. There are several hypotheses that explain the biological role of WSCPs, one of them proposing that they function as a scavenger of Chls set free upon plant senescence or pathogen attack. The biochemical properties of WSCP described in this paper are consistent with the protein acting as an efficient and flexible Chl scavenger. At limiting Chl concentrations, the recombinant WSCP apoprotein binds substoichiometric amounts of Chl (two Chls per tetramer) to form complexes that are as stable toward thermal dissociation, denaturation, and photodamage as the fully pigmented ones. If more Chl is added, these two-Chl complexes can bind another two Chls to reach the fully pigmented state. The protection of WSCP Chls against photodamage has been attributed to the apoprotein serving as a diffusion barrier for oxygen, preventing its access to triplet excited Chls and, thus, the formation of singlet oxygen. By contrast, the sequential binding of Chls by WSCP suggests a partially open or at least flexible structure, raising the question of how WSCP photoprotects its Chls without the help of carotenoids.

  3. A lactobacillus rhamnosus GG-derived soluble protein, p40, stimulates ligand release from intestinal epithelial cells to transactivate epidermal growth factor receptor

    Science.gov (United States)

    Protein p40, a Lactobacillus rhamnosus GG (LGG)-derived soluble protein, ameliorates intestinal injury and colitis, reduces apoptosis and preserves barrier function by activation of EGF receptor (EGFR) in intestinal epithelial cells. The aim of this study was to determine the mechanisms by which p40...

  4. Coregulation of soybean vegetative storage protein gene expression by methyl jasmonate and soluble sugars.

    Science.gov (United States)

    Mason, H S; Dewald, D B; Creelman, R A; Mullet, J E

    1992-03-01

    The soybean vegetative storage protein genes vspA and vspB are highly expressed in developing leaves, stems, flowers, and pods as compared with roots, seeds, and mature leaves and stems. In this paper, we report that physiological levels of methyl jasmonate (MeJA) and soluble sugars synergistically stimulate accumulation of vsp mRNAs. Treatment of excised mature soybean (Glycine max Merr. cv Williams) leaves with 0.2 molar sucrose and 10 micromolar MeJA caused a large accumulation of vsp mRNAs, whereas little accumulation occurred when these compounds were supplied separately. In soybean cell suspension cultures, the synergistic effect of sucrose and MeJA on the accumulation of vspB mRNA was maximal at 58 millimolar sucrose and was observed with fructose or glucose substituted for sucrose. In dark-grown soybean seedlings, the highest levels of vsp mRNAs occurred in the hypocotyl hook, which also contained high levels of MeJA and soluble sugars. Lower levels of vsp mRNAs, MeJA, and soluble sugars were found in the cotyledons, roots, and nongrowing regions of the stem. Wounding of mature soybean leaves induced a large accumulation of vsp mRNAs when wounded plants were incubated in the light. Wounded plants kept in the dark or illuminated plants sprayed with dichlorophenyldimethylurea, an inhibitor of photosynthetic electron transport, showed a greatly reduced accumulation of vsp mRNAs. The time courses for the accumulation of vsp mRNAs induced by wounding or sucrose/MeJA treatment were similar. These results strongly suggest that vsp expression is coregulated by endogenous levels of MeJA (or jasmonic acid) and soluble carbohydrate during normal vegetative development and in wounded leaves.

  5. A spectroscopic and photometric study of MSP companions in Galactic Globular Clusters

    OpenAIRE

    Cocozza, Gabriele

    2008-01-01

    This Thesis is devoted to the study of the optical companions of Millisecond Pulsars in Galactic Globular Clusters (GCs) as a part of a large project started at the Department of Astronomy of the Bologna University, in collaboration with other institutions (Astronomical Observatory of Cagliari and Bologna, University of Virginia), specifically dedicated to the study of the environmental effects on passive stellar evolution in galactic GCs. Globular Clusters are very efficien...

  6. Fermentation of rapeseed meal, sunflower meal and faba beans in combination with wheat bran increases solubility of protein and phosphorus

    DEFF Research Database (Denmark)

    Poulsen, Hanne Damgaard; Blaabjerg, Karoline

    2017-01-01

    BACKGROUND To increase self-supply of protein and phosphorus (P) in European pig and poultry diets and reduce nitrogen (N) and P excretion, attention is directed to approaches increasing protein and P digestibility of rapeseed, sunflower and faba beans. Wheat bran is rich in enzymes degrading...... and solubilizing protein and phytate. Herein, solubilization of protein, N and P was investigated when increasing ratios of wheat bran were fermented with rapeseed meal (RSM), sunflower meal (SFM), faba beans (FB) or a combination of these (RSM/SFM/FB). RESULTS Protein, N and P solubility was greater, for all...

  7. Structural and functional characterization of the recombinant death domain from death-associated protein kinase.

    Science.gov (United States)

    Dioletis, Evangelos; Dingley, Andrew J; Driscoll, Paul C

    2013-01-01

    Death-associated protein kinase (DAPk) is a calcium/calmodulin-regulated Ser/Thr-protein kinase that functions at an important point of integration for cell death signaling pathways. DAPk has a structurally unique multi-domain architecture, including a C-terminally positioned death domain (DD) that is a positive regulator of DAPk activity. In this study, recombinant DAPk-DD was observed to aggregate readily and could not be prepared in sufficient yield for structural analysis. However, DAPk-DD could be obtained as a soluble protein in the form of a translational fusion protein with the B1 domain of streptococcal protein G. In contrast to other DDs that adopt the canonical six amphipathic α-helices arranged in a compact fold, the DAPk-DD was found to possess surprisingly low regular secondary structure content and an absence of a stable globular fold, as determined by circular dichroism (CD), NMR spectroscopy and a temperature-dependent fluorescence assay. Furthermore, we measured the in vitro interaction between extracellular-regulated kinase-2 (ERK2) and various recombinant DAPk-DD constructs. Despite the low level of structural order, the recombinant DAPk-DD retained the ability to interact with ERK2 in a 1∶1 ratio with a K d in the low micromolar range. Only the full-length DAPk-DD could bind ERK2, indicating that the apparent 'D-motif' located in the putative sixth helix of DAPk-DD is not sufficient for ERK2 recognition. CD analysis revealed that binding of DAPk-DD to ERK2 is not accompanied by a significant change in secondary structure. Taken together our data argue that the DAPk-DD, when expressed in isolation, does not adopt a classical DD fold, yet in this state retains the capacity to interact with at least one of its binding partners. The lack of a stable globular structure for the DAPk-DD may reflect either that its folding would be supported by interactions absent in our experimental set-up, or a limitation in the structural bioinformatics

  8. Is age really the second parameter in globular clusters?

    International Nuclear Information System (INIS)

    Vandenberg, D.A.; Durrell, P.R.

    1990-01-01

    From the close similarity of the magnitude difference between the tip of the red giant branch and the turnoff in the Fe/H = about -1.3 globular cluster NGC 288, NGC 362, and M5, it is inferred that the ages of these three systems (and Palomar 5, whose horizonal branch is used to define its distance relative to the others) are not detectably different. An identical conclusion, by similar means, is reached for the Fe/H = about -2.1 globular clusters M15, M30, M68, and M92. Several recent claims that age is responsible for the wide variation in horizontal-branch morphology among clusters of the same metal abundance are not supported. 73 refs

  9. Ruprecht 106 - A young metal-poor Galactic globular cluster

    International Nuclear Information System (INIS)

    Buonanno, R.; Buscema, G.; Fusi Pecci, F.; Richer, H.B.; Fahlman, G.G.

    1990-01-01

    The first CCD photometric survey in the Galactic globular cluster Ruprecht 106 has been performed. The results show that Ruprecht 106 is a metal-poor cluster with (Fe/H) about -2 located at about 25 kpc from the Galactic center. A sizable, high centrally concentrated population of blue stragglers was detected. Significant differences in the positions of the turnoffs in the color-magnitude diagram are found compared to those in metal-poor clusters. The cluster appears younger than other typical metal-poor Galactic globulars by about 4-5 Gyr; if true, this object would represent the first direct proof of the existence of a significant age spread among old, very metal-poor clusters. 51 refs

  10. Color maps of X-ray globular clusters

    International Nuclear Information System (INIS)

    Bailyn, C.D.; Grindlay, J.E.; Cohn, H.; Lugger, P.M.

    1988-01-01

    The results of a search for optical counterparts to X-ray sources in six globular clusters, 47 Tuc, NGC 1851, NGC 6441, NGC 6624, NGC 6712, and M15, are reported. Maps of the U-B color of the central regions of the clusters were prepared. A candidate for the optical counterpart of the source in NGC 6712 was found, along with a blue region near the X-ray source in 47 Tuc. Upper limits on the colors and magnitudes of possible optical counterparts are reported for the other three clusters. The use of color maps to determine color gradients in globular clusters is explored. It is found that, while such gradients do exist and vary from cluster to cluster, they can be explained by crowding effects. Crude limits are placed on the excess populations of blue objects such as CVs, which have been postulated to be concentrated in the centers of dense clusters. 32 references

  11. Human diploid fibroblasts have receptors for the globular domain of C1Q

    International Nuclear Information System (INIS)

    Bordin, S.; Page, R.C.

    1986-01-01

    The authors showed that mass cultures of fibroblasts grown from gingival explants in DB medium with 10% human serum are enriched in a phenotype that binds C1q with an affinity much higher than the rest of the population. Because of potential biologic importance of C1q receptors, the authors studied whether the interaction between C1q and this phenotype was mediated by the globular or collagenous domains of the molecule. Globular fragments were prepared by digesting C1q with collagenase, and collagenous fragments obtained after pepsin treatment. C1q binding on cells in suspension was determined by reaction with 125 I-C1q as reported. Competition experiments were performed under conditions in which intact 125 I-C1q binding saturated all available receptors. The results showed that collagenous fragments inhibited 20% of the 125 I-C1q binding to high affinity receptors, whereas inhibition by globular fragments was 70%. Unlabeled intact C1q and collagen type 1 were used as controls, and inhibited 92% and 17% of C1q binding, respectively. These studies show that C1q interacts with the fibroblast phenotype expressing high affinity receptors through its globular domain. The authors suggest that at sites of trauma, native C1 may bind to the surface of these cells via the globular domain of C1q, and that this unique phenotype may play an important role in tissue repair

  12. Co-expression of sulphydryl oxidase and protein disulphide isomerase in Escherichia coli allows for production of soluble CRM197

    CSIR Research Space (South Africa)

    Roth, Robyn L

    2017-04-01

    Full Text Available The aim of this article is to investigate the production of soluble cross-reacting material 197 (CRM(sub197)) in Escherichia coli, a safe and effective T-cell-dependent protein carrier for polysaccharides used in the manufacture and application...

  13. Keck Spectroscopy of Globular Clusters in the Elliptical Galaxy NGC 3610

    OpenAIRE

    Strader, Jay; Brodie, Jean P.; Schweizer, Francois; Larsen, Soeren S.; Seitzer, Patrick

    2002-01-01

    We present moderate-resolution Keck spectra of nine candidate globular clusters in the possible merger-remnant elliptical galaxy NGC 3610. Eight of the objects appear to be bona fide globular clusters of NGC 3610. We find that two of the clusters belong to an old metal-poor population, five to an old metal-rich population, and only one to an intermediate-age metal-rich population. The estimated age of the intermediate-age cluster is 1-5 Gyr, which is in agreement with earlier estimates of the...

  14. Interplay between Peptide Bond Geometrical Parameters in Nonglobular Structural Contexts

    Directory of Open Access Journals (Sweden)

    Luciana Esposito

    2013-01-01

    Full Text Available Several investigations performed in the last two decades have unveiled that geometrical parameters of protein backbone show a remarkable variability. Although these studies have provided interesting insights into one of the basic aspects of protein structure, they have been conducted on globular and water-soluble proteins. We report here a detailed analysis of backbone geometrical parameters in nonglobular proteins/peptides. We considered membrane proteins and two distinct fibrous systems (amyloid-forming and collagen-like peptides. Present data show that in these systems the local conformation plays a major role in dictating the amplitude of the bond angle N-Cα-C and the propensity of the peptide bond to adopt planar/nonplanar states. Since the trends detected here are in line with the concept of the mutual influence of local geometry and conformation previously established for globular and water-soluble proteins, our analysis demonstrates that the interplay of backbone geometrical parameters is an intrinsic and general property of protein/peptide structures that is preserved also in nonglobular contexts. For amyloid-forming peptides significant distortions of the N-Cα-C bond angle, indicative of sterical hidden strain, may occur in correspondence with side chain interdigitation. The correlation between the dihedral angles Δω/ψ in collagen-like models may have interesting implications for triple helix stability.

  15. Interplay between peptide bond geometrical parameters in nonglobular structural contexts.

    Science.gov (United States)

    Esposito, Luciana; Balasco, Nicole; De Simone, Alfonso; Berisio, Rita; Vitagliano, Luigi

    2013-01-01

    Several investigations performed in the last two decades have unveiled that geometrical parameters of protein backbone show a remarkable variability. Although these studies have provided interesting insights into one of the basic aspects of protein structure, they have been conducted on globular and water-soluble proteins. We report here a detailed analysis of backbone geometrical parameters in nonglobular proteins/peptides. We considered membrane proteins and two distinct fibrous systems (amyloid-forming and collagen-like peptides). Present data show that in these systems the local conformation plays a major role in dictating the amplitude of the bond angle N-C(α)-C and the propensity of the peptide bond to adopt planar/nonplanar states. Since the trends detected here are in line with the concept of the mutual influence of local geometry and conformation previously established for globular and water-soluble proteins, our analysis demonstrates that the interplay of backbone geometrical parameters is an intrinsic and general property of protein/peptide structures that is preserved also in nonglobular contexts. For amyloid-forming peptides significant distortions of the N-C(α)-C bond angle, indicative of sterical hidden strain, may occur in correspondence with side chain interdigitation. The correlation between the dihedral angles Δω/ψ in collagen-like models may have interesting implications for triple helix stability.

  16. Structure and non-structure of centrosomal proteins.

    Science.gov (United States)

    Dos Santos, Helena G; Abia, David; Janowski, Robert; Mortuza, Gulnahar; Bertero, Michela G; Boutin, Maïlys; Guarín, Nayibe; Méndez-Giraldez, Raúl; Nuñez, Alfonso; Pedrero, Juan G; Redondo, Pilar; Sanz, María; Speroni, Silvia; Teichert, Florian; Bruix, Marta; Carazo, José M; Gonzalez, Cayetano; Reina, José; Valpuesta, José M; Vernos, Isabelle; Zabala, Juan C; Montoya, Guillermo; Coll, Miquel; Bastolla, Ugo; Serrano, Luis

    2013-01-01

    Here we perform a large-scale study of the structural properties and the expression of proteins that constitute the human Centrosome. Centrosomal proteins tend to be larger than generic human proteins (control set), since their genes contain in average more exons (20.3 versus 14.6). They are rich in predicted disordered regions, which cover 57% of their length, compared to 39% in the general human proteome. They also contain several regions that are dually predicted to be disordered and coiled-coil at the same time: 55 proteins (15%) contain disordered and coiled-coil fragments that cover more than 20% of their length. Helices prevail over strands in regions homologous to known structures (47% predicted helical residues against 17% predicted as strands), and even more in the whole centrosomal proteome (52% against 7%), while for control human proteins 34.5% of the residues are predicted as helical and 12.8% are predicted as strands. This difference is mainly due to residues predicted as disordered and helical (30% in centrosomal and 9.4% in control proteins), which may correspond to alpha-helix forming molecular recognition features (α-MoRFs). We performed expression assays for 120 full-length centrosomal proteins and 72 domain constructs that we have predicted to be globular. These full-length proteins are often insoluble: Only 39 out of 120 expressed proteins (32%) and 19 out of 72 domains (26%) were soluble. We built or retrieved structural models for 277 out of 361 human proteins whose centrosomal localization has been experimentally verified. We could not find any suitable structural template with more than 20% sequence identity for 84 centrosomal proteins (23%), for which around 74% of the residues are predicted to be disordered or coiled-coils. The three-dimensional models that we built are available at http://ub.cbm.uam.es/centrosome/models/index.php.

  17. Decreased UV light resistance of spores of Bacillus subtilis strains deficient in pyrimidine dimer repair and small, acid-soluble spore proteins

    International Nuclear Information System (INIS)

    Setlow, B.; Setlow, P.

    1988-01-01

    Loss of small, acid-soluble spore protein alpha reduced spore UV resistance 30- to 50-fold in Bacillus subtilis strains deficient in pyrimidine dimer repair, but gave only a 5- to 8-fold reduction in UV resistance in repair-proficient strains. However, both repair-proficient and -deficient spores lacking this protein had identical heat and gamma-radiation resistance

  18. Speckle imaging of globular clusters

    International Nuclear Information System (INIS)

    Sams, B.J. III

    1990-01-01

    Speckle imaging is a powerful tool for high resolution astronomy. Its application to the core regions of globular clusters produces high resolution stellar maps of the bright stars, but is unable to image the faint stars which are most reliable dynamical indicators. The limits on resolving these faint, extended objects are physical, not algorithmic, and cannot be overcome using speckle. High resolution maps may be useful for resolving multicomponent stellar systems in the cluster centers. 30 refs

  19. Globular cluster seeding by primordial black hole population

    Energy Technology Data Exchange (ETDEWEB)

    Dolgov, A. [ITEP, Bol. Cheremushkinsaya ul., 25, 117218 Moscow (Russian Federation); Postnov, K., E-mail: dolgov@fe.infn.it, E-mail: kpostnov@gmail.com [Sternberg Astronomical Institute, Moscow M.V. Lomonosov State University, Universitetskij pr., 13, Moscow 119234 (Russian Federation)

    2017-04-01

    Primordial black holes (PBHs) that form in the early Universe in the modified Affleck-Dine (AD) mechanism of baryogenesis should have intrinsic log-normal mass distribution of PBHs. We show that the parameters of this distribution adjusted to provide the required spatial density of massive seeds (≥ 10{sup 4} M {sub ⊙}) for early galaxy formation and not violating the dark matter density constraints, predict the existence of the population of intermediate-mass PBHs with a number density of 0∼ 100 Mpc{sup −3}. We argue that the population of intermediate-mass AD PBHs can also seed the formation of globular clusters in galaxies. In this scenario, each globular cluster should host an intermediate-mass black hole with a mass of a few thousand solar masses, and should not obligatorily be immersed in a massive dark matter halo.

  20. Changes in the protein profile of Habanero pepper ( Capsicum ...

    African Journals Online (AJOL)

    Protein profile was studied during the development of Capsicum chinense somatic embryos. The total protein content and profile of polypeptides (by sodium dodecyl sulfate polyacrylamide gel electrophoresis) of somatic embryos at different developmental stages (globular, heart-shaped, torpedo and cotyledonary stages) ...

  1. A tensegrity model for hydrogen bond networks in proteins.

    Science.gov (United States)

    Bywater, Robert P

    2017-05-01

    Hydrogen-bonding networks in proteins considered as structural tensile elements are in balance separately from any other stabilising interactions that may be in operation. The hydrogen bond arrangement in the network is reminiscent of tensegrity structures in architecture and sculpture. Tensegrity has been discussed before in cells and tissues and in proteins. In contrast to previous work only hydrogen bonds are studied here. The other interactions within proteins are either much stronger - covalent bonds connecting the atoms in the molecular skeleton or weaker forces like the so-called hydrophobic interactions. It has been demonstrated that the latter operate independently from hydrogen bonds. Each category of interaction must, if the protein is to have a stable structure, balance out. The hypothesis here is that the entire hydrogen bond network is in balance without any compensating contributions from other types of interaction. For sidechain-sidechain, sidechain-backbone and backbone-backbone hydrogen bonds in proteins, tensegrity balance ("closure") is required over the entire length of the polypeptide chain that defines individually folding units in globular proteins ("domains") as well as within the repeating elements in fibrous proteins that consist of extended chain structures. There is no closure to be found in extended structures that do not have repeating elements. This suggests an explanation as to why globular domains, as well as the repeat units in fibrous proteins, have to have a defined number of residues. Apart from networks of sidechain-sidechain hydrogen bonds there are certain key points at which this closure is achieved in the sidechain-backbone hydrogen bonds and these are associated with demarcation points at the start or end of stretches of secondary structure. Together, these three categories of hydrogen bond achieve the closure that is necessary for the stability of globular protein domains as well as repeating elements in fibrous proteins.

  2. Stabilization of apoglobin by low temperature increases yield of soluble recombinant hemoglobin in Escherichia coli.

    Science.gov (United States)

    Weickert, M J; Pagratis, M; Curry, S R; Blackmore, R

    1997-01-01

    Accumulation of soluble recombinant hemoglobin (rHb1.1) in Escherichia coli requires proper protein folding, prosthetic group (heme) addition, and subunit assembly. This served as a new model system for the study of the effects of temperature, protein synthesis rates, and protein accumulation rates on protein solubility in E. coli. Fermentation expression of rHb1.1 at 30 degrees C from cultures containing a medium or high globin gene dosage (pBR-based or pUC-based plasmids with rHb1.1 genes under the control of the tac promoter) was compared. A medium gene dosage resulted in rHb1.1 accumulating to approximately 7% of the soluble cell protein, of which 78% was soluble. A high globin gene dosage resulted in a > or = 3-fold increase in total globin to 23 to 24% of the soluble cell protein, but 70% was insoluble. Accumulation of insoluble rHb1.1 began immediately upon induction. The proportion of rHb1.1 from the high globin gene dosage that accumulated as insoluble globin was affected by reducing (i) the inducer concentration and (ii) the temperature. Reducing the inducer concentration reduced globin synthesis up to eightfold but increased the proportion of soluble rHb1.1 to 93%. In contrast, total globin protein synthesis was barely affected by reducing the temperature from 30 to 26 degrees C, while soluble globin accumulation increased > 2-fold to approximately 15% of the soluble cell protein. The contrast between the effects of reducing rates of protein synthesis and accumulation and those of reducing temperature suggests that lower temperature stabilizes one or more folding intermediates. We propose a simplified physical model which integrates protein synthesis, folding, and heme association. This model shows that temperature-dependent apoglobin stability is the most critical factor in soluble rHb1.1 accumulation. PMID:9361418

  3. Globular adiponectin controls insulin-mediated vasoreactivity in muscle through AMPKα2

    DEFF Research Database (Denmark)

    de Boer, Michiel P; Meijer, Rick I; Richter, Erik

    2016-01-01

    Decreased tissue perfusion increases the risk of developing insulin resistance and cardiovascular disease in obesity, and decreased levels of globular adiponectin (gAdn) have been proposed to contribute to this risk. We hypothesized that gAdn controls insulin's vasoactive effects through AMP......-activated protein kinase (AMPK), specifically its α2 subunit, and studied the mechanisms involved. In healthy volunteers, we found that decreased plasma gAdn levels in obese subjects associate with insulin resistance and reduced capillary perfusion during hyperinsulinemia. In cultured human microvascular...... endothelial cells (HMEC), gAdn increased AMPK activity. In isolated muscle resistance arteries gAdn uncovered insulin-induced vasodilation by selectively inhibiting insulin-induced activation of ERK1/2, and the AMPK inhibitor compound C as well as genetic deletion of AMPKα2 blunted insulin...

  4. Effect of electrical stunning frequency on meat quality, plasma parameters, and protein solubility of broilers.

    Science.gov (United States)

    Huang, J C; Yang, J; Zhang, B H; Huang, M; Chen, K J; Xu, X L; Zhou, G H

    2017-08-01

    This study was designed to compare the effects of different stunning frequencies of pulsed direct current on meat quality of broilers. This was achieved by investigating plasma parameters, blood loss, carcass damage, meat water-holding capacity, meat color, meat shear value, muscle pH, and protein solubility. A total of 400 broilers was divided into 5 treatment groups and stunned with 500, 600, 700, 800, and 900 Hz at 15 V for 10 seconds. Blood samples were collected immediately after cutting the neck. Pectoralis major muscles were removed from the carcass after chilling and placed in ice. Breast muscle pH and meat color were determined at both 2 and 24 h postmortem. Drip loss, cooking loss, pressing loss, and cooked breast meat-shear values were determined at 24 h postmortem. Treatment at 500 and 900 Hz significantly increased (P meat color were not affected by stunning frequency. In the 500 and 900 Hz groups, the protein solubility and shear force values were significantly lower (P < 0.05) and drip loss was significantly higher (P < 0.05) than in the 700 Hz group. This study indicates that the waveform of the pulsed direct current is acceptable for stunning broilers at a stunning frequency of 700 Hz. © 2017 Poultry Science Association Inc.

  5. New bound on neutrino dipole moments from globular-cluster stars

    Science.gov (United States)

    Raffelt, Georg G.

    1990-01-01

    Neutrino dipole moments mu(nu) would increase the core mass of red giants at the helium flash by delta(Mc) = 0.015 solar mass x mu(nu)/10 to the -12th muB (where muB is the Bohr magneton) because of enhanced neutrino losses. Existing measurements of the bolometric magnitudes of the brightest red giants in 26 globular clusters, number counts of horizontal-branch stars and red giants in 15 globular clusters, and statistical parallax determinations of field RR Lyr luminosities yield delta(Mc) = 0.009 + or - 0.012 solar mass, so that conservatively mu(nu) is less than 3 x 10 to the -12th muB.

  6. Efficient expression of a soluble lipid transfer protein (LTP) of Platanus orientalis using short peptide tags and structural comparison with the natural form.

    Science.gov (United States)

    Salari, Farhad; Vahedi, Fatemeh; Chamani, Jamshidkhan; Varasteh, Abdolreza; Ketabdar, Hanieh; Sankian, Mojtaba

    2015-01-01

    Successful recombinant allergen-based immunotherapy has drawn a great deal of attention to use recombinant allergens for new therapeutic and/or diagnostic strategies. The Escherichia coli expression system is frequently used to produce recombinant allergens; however, protein expression in E. coli often results in inclusion bodies. Here, we focused on the expression of two recombinant soluble forms of Pla or 3 using solubility-enhancing peptide tags, human immune deficiency virus type 1 transactivator of transcription core domain and poly-arginine-lysine: rTAT-Pla or 3 and rPoly-Arg-Lys-Pla or 3. Structural characteristics and IgE reactivity of purified recombinant proteins were compared with natural Pla or 3 (nPla or 3) isolated from Platanus orientalis using circular dichroism spectra, fluorescence spectroscopy, and immunoblotting. Likewise, intrinsic viscosity and Stokes radius of the natural and recombinant Pla or 3 allergens were determined to analyze structural compactness in aqueous media. The results indicate high-level solubility and efficient expression of the fusion proteins (rTAT-Pla or 3 and rPoly-Arg-Lys-Pla or 3) compared with the wild-type recombinant. Furthermore, the similar structural characteristics and IgE-binding activities of the fusion proteins to nPla or 3 provide a promising tool for allergy diagnosis and treatment. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  7. Scoring function to predict solubility mutagenesis

    Directory of Open Access Journals (Sweden)

    Deutsch Christopher

    2010-10-01

    Full Text Available Abstract Background Mutagenesis is commonly used to engineer proteins with desirable properties not present in the wild type (WT protein, such as increased or decreased stability, reactivity, or solubility. Experimentalists often have to choose a small subset of mutations from a large number of candidates to obtain the desired change, and computational techniques are invaluable to make the choices. While several such methods have been proposed to predict stability and reactivity mutagenesis, solubility has not received much attention. Results We use concepts from computational geometry to define a three body scoring function that predicts the change in protein solubility due to mutations. The scoring function captures both sequence and structure information. By exploring the literature, we have assembled a substantial database of 137 single- and multiple-point solubility mutations. Our database is the largest such collection with structural information known so far. We optimize the scoring function using linear programming (LP methods to derive its weights based on training. Starting with default values of 1, we find weights in the range [0,2] so that predictions of increase or decrease in solubility are optimized. We compare the LP method to the standard machine learning techniques of support vector machines (SVM and the Lasso. Using statistics for leave-one-out (LOO, 10-fold, and 3-fold cross validations (CV for training and prediction, we demonstrate that the LP method performs the best overall. For the LOOCV, the LP method has an overall accuracy of 81%. Availability Executables of programs, tables of weights, and datasets of mutants are available from the following web page: http://www.wsu.edu/~kbala/OptSolMut.html.

  8. MOCK OBSERVATIONS OF BLUE STRAGGLERS IN GLOBULAR CLUSTER MODELS

    International Nuclear Information System (INIS)

    Sills, Alison; Glebbeek, Evert; Chatterjee, Sourav; Rasio, Frederic A.

    2013-01-01

    We created artificial color-magnitude diagrams of Monte Carlo dynamical models of globular clusters and then used observational methods to determine the number of blue stragglers in those clusters. We compared these blue stragglers to various cluster properties, mimicking work that has been done for blue stragglers in Milky Way globular clusters to determine the dominant formation mechanism(s) of this unusual stellar population. We find that a mass-based prescription for selecting blue stragglers will select approximately twice as many blue stragglers than a selection criterion that was developed for observations of real clusters. However, the two numbers of blue stragglers are well-correlated, so either selection criterion can be used to characterize the blue straggler population of a cluster. We confirm previous results that the simplified prescription for the evolution of a collision or merger product in the BSE code overestimates their lifetimes. We show that our model blue stragglers follow similar trends with cluster properties (core mass, binary fraction, total mass, collision rate) as the true Milky Way blue stragglers as long as we restrict ourselves to model clusters with an initial binary fraction higher than 5%. We also show that, in contrast to earlier work, the number of blue stragglers in the cluster core does have a weak dependence on the collisional parameter Γ in both our models and in Milky Way globular clusters

  9. Discovery of a ~205 Hz X-ray pulsar in the globular cluster NGC 6440

    NARCIS (Netherlands)

    Altamirano, D.; Strohmayer, T.E.; Heinke, C.O.; Markwardt, C.B.; Swank, J.H.; Pereira, D.; Smith, E.; Wijnands, R.; Linares, M.; Patruno, A.; Casella, P.; van der Klis, M.

    2009-01-01

    Discovery of a 205 Hz X-ray pulsar in the globular cluster NGC 6440 The globular cluster NGC 6440 was observed by the PCA instrument aboard RXTE on August 30, 2009 at 01:42 (UTC). The observation lasted for approximately 3000 seconds and the source was detected with an intensity of ~7 mCrab (2-10

  10. VARIABLE STARS IN LARGE MAGELLANIC CLOUD GLOBULAR CLUSTERS. II. NGC 1786

    International Nuclear Information System (INIS)

    Kuehn, Charles A.; Smith, Horace A.; De Lee, Nathan; Catelan, Márcio; Pritzl, Barton J.; Borissova, Jura

    2012-01-01

    This is the second in a series of papers studying the variable stars in Large Magellanic Cloud globular clusters. The primary goal of this series is to study how RR Lyrae stars in Oosterhoff-intermediate systems compare to their counterparts in Oosterhoff I/II systems. In this paper, we present the results of our new time-series B–V photometric study of the globular cluster NGC 1786. A total of 65 variable stars were identified in our field of view. These variables include 53 RR Lyraes (27 RRab, 18 RRc, and 8 RRd), 3 classical Cepheids, 1 Type II Cepheid, 1 Anomalous Cepheid, 2 eclipsing binaries, 3 Delta Scuti/SX Phoenicis variables, and 2 variables of undetermined type. Photometric parameters for these variables are presented. We present physical properties for some of the RR Lyrae stars, derived from Fourier analysis of their light curves. We discuss several different indicators of Oosterhoff type which indicate that the Oosterhoff classification of NGC 1786 is not as clear cut as what is seen in most globular clusters.

  11. A 5.75-millisecond pulsar in the globular cluster 47 Tucanae

    International Nuclear Information System (INIS)

    Manchester, R.N.; Lyne, A.G.; Johnston, S.; D'Amico, N.; Lim, J.; Kniffen, D.A.

    1990-01-01

    Millisecond pulsars are generally believed to be old pulsars that have been spun up ('recycled') as a result of accretion of matter from a companion in a low-mass X-ray binary system. As there is a high incidence of such systems in globular clusters, these are good places to search for millisecond pulsars; so far, ten globular-cluster pulsars have been detected unambiguously. Using the Parkes radiotelescope in Australia, we have found a pulsar with a period of 5.75 ms and a dispersion measure of 25 cm -3 pc in the direction of 47 Tucanae. Despite its probable origin as a member of a binary system, timing measurements show that the pulsar is now single. The observed dispersion measure is consistent with the pulsar lying outside the galactic electron layer and within 47 Tucanae; but it is very different from the value of 67 cm -3 pc for the pulsars that were reported recently as being in this globular cluster, and we suggest that the latter pulsars probably do not in fact lie within 47 Tucanae. (author)

  12. THE SIZE DIFFERENCE BETWEEN RED AND BLUE GLOBULAR CLUSTERS IS NOT DUE TO PROJECTION EFFECTS

    International Nuclear Information System (INIS)

    Webb, Jeremy J.; Harris, William E.; Sills, Alison

    2012-01-01

    Metal-rich (red) globular clusters in massive galaxies are, on average, smaller than metal-poor (blue) globular clusters. One of the possible explanations for this phenomenon is that the two populations of clusters have different spatial distributions. We test this idea by comparing clusters observed in unusually deep, high signal-to-noise images of M87 with a simulated globular cluster population in which the red and blue clusters have different spatial distributions, matching the observations. We compare the overall distribution of cluster effective radii as well as the relationship between effective radius and galactocentric distance for both the observed and simulated red and blue sub-populations. We find that the different spatial distributions does not produce a significant size difference between the red and blue sub-populations as a whole or at a given galactocentric distance. These results suggest that the size difference between red and blue globular clusters is likely due to differences during formation or later evolution.

  13. The Size Difference between Red and Blue Globular Clusters is not due to Projection Effects

    Science.gov (United States)

    Webb, Jeremy J.; Harris, William E.; Sills, Alison

    2012-11-01

    Metal-rich (red) globular clusters in massive galaxies are, on average, smaller than metal-poor (blue) globular clusters. One of the possible explanations for this phenomenon is that the two populations of clusters have different spatial distributions. We test this idea by comparing clusters observed in unusually deep, high signal-to-noise images of M87 with a simulated globular cluster population in which the red and blue clusters have different spatial distributions, matching the observations. We compare the overall distribution of cluster effective radii as well as the relationship between effective radius and galactocentric distance for both the observed and simulated red and blue sub-populations. We find that the different spatial distributions does not produce a significant size difference between the red and blue sub-populations as a whole or at a given galactocentric distance. These results suggest that the size difference between red and blue globular clusters is likely due to differences during formation or later evolution.

  14. Overcoming the solubility limit with solubility-enhancement tags: successful applications in biomolecular NMR studies

    International Nuclear Information System (INIS)

    Zhou Pei; Wagner, Gerhard

    2010-01-01

    Although the rapid progress of NMR technology has significantly expanded the range of NMR-trackable systems, preparation of NMR-suitable samples that are highly soluble and stable remains a bottleneck for studies of many biological systems. The application of solubility-enhancement tags (SETs) has been highly effective in overcoming solubility and sample stability issues and has enabled structural studies of important biological systems previously deemed unapproachable by solution NMR techniques. In this review, we provide a brief survey of the development and successful applications of the SET strategy in biomolecular NMR. We also comment on the criteria for choosing optimal SETs, such as for differently charged target proteins, and recent new developments on NMR-invisible SETs.

  15. Effect of urdbean leaf crinkle virus infection on total soluble protein and antioxidant enzymes in blackgram plants

    International Nuclear Information System (INIS)

    Ashfaq, M.; Mughal, S.M.; Khan, A.; Javed, N.; Sahi, S.T.; Shahid, M.

    2010-01-01

    Urdbean leaf crinkle virus (ULCV) is a common, wide spread, destructive and economically important disease causing systemic infection in blackgram (Vigna mungo (L.) Hepper), resulting in extreme crinkling, curling, puckering and rugosity of leaves, and yield reductions. Effect of viral infection was investigated on total soluble proteins and antioxidant enzymes activity in two genotypes viz., Mash-88-susceptible and CM-2002-resistant, at different growth stages under both the inoculated and un-inoculated conditions. ULCV infection resulted in significant increase in total soluble protein contents of the leaves in both genotypes. In healthy plant, super oxide dismutase (SOD), catalase (CAT) and peroxidase (PO) showed similar activity levels. In inoculated plants of Mash-88, SOD and PO activities decreased and increased non-significantly at all growth stages, respectively. The activities of PO and SOD increased and decreased significantly after 15 and 30 days of inoculation in resistant genotype, respectively. No significant changes in catalase (CAT) activity were detected in ULCV-infected leaves over the control. It was concluded that the super oxide dismutase and peroxidases might be associated with resistance/susceptibility to ULCV infection. (author)

  16. Bee venom phospholipase A2 as a membrane-binding vector for cell surface display or internalization of soluble proteins.

    Science.gov (United States)

    Babon, Aurélie; Wurceldorf, Thibault; Almunia, Christine; Pichard, Sylvain; Chenal, Alexandre; Buhot, Cécile; Beaumelle, Bruno; Gillet, Daniel

    2016-06-15

    We showed that bee venom phospholipase A2 can be used as a membrane-binding vector to anchor to the surface of cells a soluble protein fused to its C-terminus. ZZ, a two-domain derivative of staphylococcal protein A capable of binding constant regions of antibodies was fused to the C-terminus of the phospholipase or to a mutant devoid of enzymatic activity. The fusion proteins bound to the surface of cells and could themselves bind IgGs. Their fate depended on the cell type to which they bound. On the A431 carcinoma cell line the proteins remained exposed on the cell surface. In contrast, on human dendritic cells the proteins were internalized into early endosomes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Synthesis of giant globular multivalent glycofullerenes as potent inhibitors in a model of Ebola virus infection

    Science.gov (United States)

    Muñoz, Antonio; Sigwalt, David; Illescas, Beatriz M.; Luczkowiak, Joanna; Rodríguez-Pérez, Laura; Nierengarten, Iwona; Holler, Michel; Remy, Jean-Serge; Buffet, Kevin; Vincent, Stéphane P.; Rojo, Javier; Delgado, Rafael; Nierengarten, Jean-François; Martín, Nazario

    2016-01-01

    The use of multivalent carbohydrate compounds to block cell-surface lectin receptors is a promising strategy to inhibit the entry of pathogens into cells and could lead to the discovery of novel antiviral agents. One of the main problems with this approach, however, is that it is difficult to make compounds of an adequate size and multivalency to mimic natural systems such as viruses. Hexakis adducts of [60]fullerene are useful building blocks in this regard because they maintain a globular shape at the same time as allowing control over the size and multivalency. Here we report water-soluble tridecafullerenes decorated with 120 peripheral carbohydrate subunits, so-called ‘superballs’, that can be synthesized efficiently from hexakis adducts of [60]fullerene in one step by using copper-catalysed azide-alkyne cycloaddition click chemistry. Infection assays show that these superballs are potent inhibitors of cell infection by an artificial Ebola virus with half-maximum inhibitory concentrations in the subnanomolar range.

  18. NGC 6273: Towards Defining A New Class of Galactic Globular Clusters?

    Science.gov (United States)

    Johnson, Christian I.; Rich, Robert Michael; Pilachowski, Catherine A.; Caldwell, Nelson; Mateo, Mario L.; Ira Bailey, John; Crane, Jeffrey D.

    2016-01-01

    A growing number of observations have found that several Galactic globular clusters exhibit abundance dispersions beyond the well-known light element (anti-)correlations. These clusters tend to be very massive, have >0.1 dex intrinsic metallicity dispersions, have complex sub-giant branch morphologies, and have correlated [Fe/H] and s-process element enhancements. Interestingly, nearly all of these clusters discovered so far have [Fe/H]~-1.7. In this context, we have examined the chemical composition of 18 red giant branch (RGB) stars in the massive, metal-poor Galactic bulge globular cluster NGC 6273 using high signal-to-noise, high resolution (R~27,000) spectra obtained with the Michigan/Magellan Fiber System (M2FS) and MSpec spectrograph mounted on the Magellan-Clay 6.5m telescope at Las Campanas Observatory. We find that the cluster exhibits a metallicity range from [Fe/H]=-1.80 to -1.30 and is composed of two dominant populations separated in [Fe/H] and [La/Fe] abundance. The increase in [La/Eu] as a function of [La/H] suggests that the increase in [La/Fe] with [Fe/H] is due to almost pure s-process enrichment. The most metal-rich star in our sample is not strongly La-enhanced, but is α-poor and may belong to a third "anomalous" stellar population. The two dominant populations exhibit the same [Na/Fe]-[Al/Fe] correlation found in other "normal" globular clusters. Therefore, NGC 6273 joins ω Centauri, M 22, M 2, and NGC 5286 as a possible new class of Galactic globular clusters.

  19. Cytokine-like factor-1, a novel soluble protein, shares homology with members of the cytokine type I receptor family.

    Science.gov (United States)

    Elson, G C; Graber, P; Losberger, C; Herren, S; Gretener, D; Menoud, L N; Wells, T N; Kosco-Vilbois, M H; Gauchat, J F

    1998-08-01

    In this report we describe the identification, cloning, and expression pattern of human cytokine-like factor 1 (hCLF-1) and the identification and cloning of its murine homologue. They were identified from expressed sequence tags using amino acid sequences from conserved regions of the cytokine type I receptor family. Human CLF-1 and murine CLF-1 shared 96% amino acid identity and significant homology with many cytokine type I receptors. CLF-1 is a secreted protein, suggesting that it is either a soluble subunit within a cytokine receptor complex, like the soluble form of the IL-6R alpha-chain, or a subunit of a multimeric cytokine, e.g., IL-12 p40. The highest levels of hCLF-1 mRNA were observed in lymph node, spleen, thymus, appendix, placenta, stomach, bone marrow, and fetal lung, with constitutive expression of CLF-1 mRNA detected in a human kidney fibroblastic cell line. In fibroblast primary cell cultures, CLF-1 mRNA was up-regulated by TNF-alpha, IL-6, and IFN-gamma. Western blot analysis of recombinant forms of hCLF-1 showed that the protein has the tendency to form covalently linked di- and tetramers. These results suggest that CLF-1 is a novel soluble cytokine receptor subunit or part of a novel cytokine complex, possibly playing a regulatory role in the immune system and during fetal development.

  20. Reduction in lipophilicity improved the solubility, plasma–protein binding, and permeability of tertiary sulfonamide RORc inverse agonists

    Energy Technology Data Exchange (ETDEWEB)

    Fauber, Benjamin P.; René, Olivier; de Leon Boenig, Gladys; Burton, Brenda; Deng, Yuzhong; Eidenschenk, Céline; Everett, Christine; Gobbi, Alberto; Hymowitz, Sarah G.; Johnson, Adam R.; La, Hank; Liimatta, Marya; Lockey, Peter; Norman, Maxine; Ouyang, Wenjun; Wang, Weiru; Wong, Harvey (Genentech); (Argenta)

    2014-08-01

    Using structure-based drug design principles, we identified opportunities to reduce the lipophilicity of our tertiary sulfonamide RORc inverse agonists. The new analogs possessed improved RORc cellular potencies with >77-fold selectivity for RORc over other nuclear receptors in our cell assay suite. The reduction in lipophilicity also led to an increased plasma–protein unbound fraction and improvements in cellular permeability and aqueous solubility.

  1. Action of mercurials on activity of partially purified soluble protein kinase C from mice brain

    International Nuclear Information System (INIS)

    Inoue, Y.; Saijoh, K.; Sumino, K.

    1988-01-01

    The enzymatic activity of soluble protein kinase C from mice brain was inhibited by mercuric chloride (II) (HgCl 2 ) and organic mercurials, i.e. methyl mercury, phenyl mercury and p-chloromercuribenzoic acid (PCMB). The IC50 was 0.08 μM for HgCl 2 and about 1 μM for organic mercurials. Sulfhydryl blocking reagents such as 5.5'-dithiobis-2-nitrobenzoic acid (DTNB) and N-ethylmaleimide (NEM) were less potent but nevertheless inhibited the enzymic activity of protein kinase C. The Hill coefficients of HgCl 2 , DTNB and NEM were close to unity whereas the values for organic mercurials were 1.3 to 1.5. The inhibition was of a non-competitive type with respect to Hl histone. 3 H-PDBu binding activity was also inhibited by all of the reagents in a non-competitive manner. Mercurials apparently bind to sulfhydryl groups of protein kinase C to inhibit the enzymatic activity. (author)

  2. [Interconnection between architecture of protein globule and disposition of conformational conservative oligopeptides in proteins from one protein family].

    Science.gov (United States)

    Batianovskiĭ, A V; Filatov, I V; Namiot, V A; Esipova, N G; Volotovskiĭ, I D

    2012-01-01

    It was shown that selective interactions between helical segments of macromolecules can realize in globular proteins in the segments characterized by the same periodicities of charge distribution i.e. between conformationally conservative oligopeptides. It was found that in the macromolecules of alpha-helical proteins conformationally conservative oligopeptides are disposed at a distance being characteristic of direct interactions. For representatives of many structural families of alpha-type proteins specific disposition of conformationally conservative segments is observed. This disposition is inherent to a particular structural family. Disposition of conformationally conservative segments is not related to homology of the amino acid sequence but reflects peculiarities of native 3D-architectures of protein globules.

  3. Soluble CD163

    DEFF Research Database (Denmark)

    Møller, Holger J

    2012-01-01

    CD163 is an endocytic receptor for haptoglobin-hemoglobin complexes and is expressed solely on macrophages and monocytes. As a result of ectodomain shedding, the extracellular portion of CD163 circulates in blood as a soluble protein (sCD163) at 0.7-3.9 mg/l in healthy individuals. The function o...

  4. Protein conformational transitions at the liquid-gas interface as studied by dilational surface rheology.

    Science.gov (United States)

    Noskov, Boris A

    2014-04-01

    Experimental results on the dynamic dilational surface elasticity of protein solutions are analyzed and compared. Short reviews of the protein behavior at the liquid-gas interface and the dilational surface rheology precede the main sections of this work. The kinetic dependencies of the surface elasticity differ strongly for the solutions of globular and non-globular proteins. In the latter case these dependencies are similar to those for solutions of non-ionic amphiphilic polymers and have local maxima corresponding to the formation of the distal region of the surface layer (type I). In the former case the dynamic surface elasticity is much higher (>60 mN/m) and the kinetic dependencies are monotonical and similar to the data for aqueous dispersions of solid nanoparticles (type II). The addition of strong denaturants to solutions of bovine serum albumin and β-lactoglobulin results in an abrupt transition from the type II to type I dependencies if the denaturant concentration exceeds a certain critical value. These results give a strong argument in favor of the preservation of the protein globular structure in the course of adsorption without any denaturants. The addition of cationic surfactants also can lead to the non-monotonical kinetic dependencies of the dynamic surface elasticity indicating destruction of the protein tertiary and secondary structures. The addition of anionic surfactants gives similar results only for the protein solutions of high ionic strength. The influence of cationic surfactants on the local maxima of the kinetic dependencies of the dynamic surface elasticity for solutions of a non-globular protein (β-casein) differs from the influence of anionic surfactants due to the heterogeneity of the charge distribution along the protein chain. In this case one can use small admixtures of ionic surfactants as probes of the adsorption mechanism. The effect of polyelectrolytes on the kinetic dependencies of the dynamic surface elasticity of protein

  5. High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Huang Yadong

    2010-02-01

    Full Text Available Abstract Background Fibroblast growth factor 21 (FGF21 is a promising drug candidate to combat metabolic diseases. However, high-level expression and purification of recombinant FGF21 (rFGF21 in Escherichia coli (E. coli is difficult because rFGF21 forms inclusion bodies in the bacteria making it difficult to purify and obtain high concentrations of bioactive rFGF21. To overcome this problem, we fused the FGF21 with SUMO (Small ubiquitin-related modifier by polymerase chain reaction (PCR, and expressed the fused gene in E. coli BL21(DE3. Results By inducing with IPTG, SUMO-FGF21 was expressed at a high level. Its concentration reached 30% of total protein, and exceeded 95% of all soluble proteins. The fused protein was purified by DEAE sepharose FF and Ni-NTA affinity chromatography. Once cleaved by the SUMO protease, the purity of rFGF21 by high performance liquid chromatography (HPLC was shown to be higher than 96% with low endotoxin level (in vivo animal experiments showed that rFGF21 produced by using this method, could decrease the concentration of plasma glucose in diabetic rats by streptozotocin (STZ injection. Conclusions This study demonstrated that SUMO, when fused with FGF21, was able to promote its soluble expression of the latter in E. coli, making it more convenient to purify rFGF21 than previously. This may be a better method to produce rFGF21 for pharmaceutical research and development.

  6. Edades relativas de cúmulos globulares

    Science.gov (United States)

    Miller Bertolami, M.; Forte, J. C.

    El trabajo de Rossemberg et al (1999), estudia las edades relativas de cúmulos globulares galácticos mediante el análisis de ciertos parámetros morfológicos de los diagramas color-magnitud de dichos cúmulos. Este trabajo se centra en tres puntos: analizar la consistencia de los resultados obtenidos por Rossemberg et al (1999) al emplear observaciones en el sistema fotométrico de Washington, más precisamente, las magnitudes C y T1 en lugar de las magnitudes V e I utilizadas por dichos autores. De la existencia de colores integrados, metalicidad y edad (relativa) para 21 de los cúmulos utilizados en dicho trabajo, se analiza la consistencia de estos resultados con las dependencias de color integrado como función de la edad y la metalicidad que se desprenden de los modelos teóricos de luz integrada por Worthey (1994), Schulz (2002) y Lee et al (2002). Por último se lleva a cabo una breve comparación de la morfología de los diagramas color-magnitud de los cúmulos globulares y de las isocronas utilizadas, a fin de intentar identificar algunas de las posibles causas de las diferencias observadas en los incisos anteriores.

  7. Theoretical stellar luminosity functions and globular cluster ages and compositions

    International Nuclear Information System (INIS)

    Ratcliff, S.J.

    1985-01-01

    The ages and chemical compositions of the stars in globular clusters are of great interest, particularly because age estimates from the well-known exercise of fitting observed color-magnitude diagrams to theoretical predictions tend to yield ages in excess of the Hubble time (an estimate to the age of the Universe) in standard cosmological models, for currently proposed high values of Hubble's constant (VandenBerg 1983). Relatively little use has been made of stellar luminosity functions of the globular clusters, for which reliable observations are now becoming available, to constrain the ages or compositions. The comparison of observed luminosity functions to theoretical ones allows one to take advantage of information not usually used, and has the advantage of being relatively insensitive to our lack of knowledge of the detailed structure of stellar envelopes and atmospheres. A computer program was developed to apply standard stellar evolutionary theory, using the most recently available input physics (opacities, nuclear reaction rates), to the calculation of the evolution of low-mass Population II stars. An algorithm for computing luminosity functions from the evolutionary tracks was applied to sets of tracks covering a broad range of chemical compositions and ages, such as may be expected for globular clusters

  8. The Observational and Theoretical Tidal Radii of Globular Clusters in M87

    Science.gov (United States)

    Webb, Jeremy J.; Sills, Alison; Harris, William E.

    2012-02-01

    Globular clusters have linear sizes (tidal radii) which theory tells us are determined by their masses and by the gravitational potential of their host galaxy. To explore the relationship between observed and expected radii, we utilize the globular cluster population of the Virgo giant M87. Unusually deep, high signal-to-noise images of M87 are used to measure the effective and limiting radii of approximately 2000 globular clusters. To compare with these observations, we simulate a globular cluster population that has the same characteristics as the observed M87 cluster population. Placing these simulated clusters in the well-studied tidal field of M87, the orbit of each cluster is solved and the theoretical tidal radius of each cluster is determined. We compare the predicted relationship between cluster size and projected galactocentric distance to observations. We find that for an isotropic distribution of cluster velocities, theoretical tidal radii are approximately equal to observed limiting radii for R gc < 10 kpc. However, the isotropic simulation predicts a steep increase in cluster size at larger radii, which is not observed in large galaxies beyond the Milky Way. To minimize the discrepancy between theory and observations, we explore the effects of orbital anisotropy on cluster sizes, and suggest a possible orbital anisotropy profile for M87 which yields a better match between theory and observations. Finally, we suggest future studies which will establish a stronger link between theoretical tidal radii and observed radii.

  9. THE OBSERVATIONAL AND THEORETICAL TIDAL RADII OF GLOBULAR CLUSTERS IN M87

    International Nuclear Information System (INIS)

    Webb, Jeremy J.; Sills, Alison; Harris, William E.

    2012-01-01

    Globular clusters have linear sizes (tidal radii) which theory tells us are determined by their masses and by the gravitational potential of their host galaxy. To explore the relationship between observed and expected radii, we utilize the globular cluster population of the Virgo giant M87. Unusually deep, high signal-to-noise images of M87 are used to measure the effective and limiting radii of approximately 2000 globular clusters. To compare with these observations, we simulate a globular cluster population that has the same characteristics as the observed M87 cluster population. Placing these simulated clusters in the well-studied tidal field of M87, the orbit of each cluster is solved and the theoretical tidal radius of each cluster is determined. We compare the predicted relationship between cluster size and projected galactocentric distance to observations. We find that for an isotropic distribution of cluster velocities, theoretical tidal radii are approximately equal to observed limiting radii for R gc < 10 kpc. However, the isotropic simulation predicts a steep increase in cluster size at larger radii, which is not observed in large galaxies beyond the Milky Way. To minimize the discrepancy between theory and observations, we explore the effects of orbital anisotropy on cluster sizes, and suggest a possible orbital anisotropy profile for M87 which yields a better match between theory and observations. Finally, we suggest future studies which will establish a stronger link between theoretical tidal radii and observed radii.

  10. Short-term X-ray variability of the globular cluster source 4U 1820 - 30 (NGC 6624)

    Science.gov (United States)

    Stella, L.; Kahn, S. M.; Grindlay, J. E.

    1984-01-01

    Analytical techniques for improved identification of the temporal and spectral variability properties of globular cluster and galactic bulge X-ray sources are described in terms of their application to a large set of observations of the source 4U 1820 - 30 in the globular cluster NGC 6624. The autocorrelation function, cross-correlations, time skewness function, erratic periodicities, and pulse trains are examined. The results are discussed in terms of current models with particular emphasis on recent accretion disk models. It is concluded that the analyzed observations provide the first evidence for shot-noise variability in a globular cluster X-ray source.

  11. Tidal stripping stellar substructures around four metal-poor globular clusters in the galactic bulge

    International Nuclear Information System (INIS)

    Chun, Sang-Hyun; Kang, Minhee; Jung, DooSeok; Sohn, Young-Jong

    2015-01-01

    We investigate the spatial density configuration of stars around four metal-poor globular clusters (NGC 6266, NGC 6626, NGC 6642, and NGC 6723) in the Galactic bulge region using wide-field deep J, H, and K imaging data obtained with the Wide Field Camera near-infrared array on the United Kingdom Infrared Telescope. A statistical weighted filtering algorithm for the stars on the color–magnitude diagram is applied in order to sort cluster member candidates from the field star contamination. In two-dimensional isodensity contour maps of the clusters, we find that all four of the globular clusters exhibit strong evidence of tidally stripped stellar features beyond the tidal radius in the form of tidal tails or small density lobes/chunks. The orientations of the extended stellar substructures are likely to be associated with the effect of dynamic interaction with the Galaxy and the cluster's space motion. The observed radial density profiles of the four globular clusters also describe the extended substructures; they depart from theoretical King and Wilson models and have an overdensity feature with a break in the slope of the profile at the outer region of clusters. The observed results could imply that four globular clusters in the Galactic bulge region have experienced strong environmental effects such as tidal forces or bulge/disk shocks of the Galaxy during the dynamical evolution of globular clusters. These observational results provide further details which add to our understanding of the evolution of clusters in the Galactic bulge region as well as the formation of the Galaxy.

  12. Testing modified gravity with globular clusters: the case of NGC 2419

    Science.gov (United States)

    Llinares, Claudio

    2018-05-01

    The dynamics of globular clusters has been studied in great detail in the context of general relativity as well as with modifications of gravity that strongly depart from the standard paradigm such as Modified Newtonian Dynamics. However, at present there are no studies that aim to test the impact that less extreme modifications of gravity (e.g. models constructed as alternatives to dark energy) have on the behaviour of globular clusters. This Letter presents fits to the velocity dispersion profile of the cluster NGC 2419 under the symmetron-modified gravity model. The data show an increase in the velocity dispersion towards the centre of the cluster which could be difficult to explain within general relativity. By finding the best-fitting solution associated with the symmetron model, we show that this tension does not exist in modified gravity. However, the best-fitting parameters give a model that is inconsistent with the dynamics of the Solar system. Exploration of different screening mechanisms should give us the chance to understand if it is possible to maintain the appealing properties of the symmetron model when it comes to globular clusters and at the same time recover the Solar system dynamics properly.

  13. Syntheses of nucleic acid and protein in somatic embryos of Fritillaria ussuriensis maxim in different development stages

    International Nuclear Information System (INIS)

    Wang Shuyu; Tang Wei; Wang Hui

    1993-09-01

    After developing a procedure for somatic embryogenesis in Fritillaria ussuriensis, dynamics on the syntheses of DNA, RNA, and protein during globular, heart-shaped, torpedo-shaped, cotyledonary, and mature somatic embryo stages was demonstrated by both autoradiography and scintillation counting. The rates of syntheses of DNA, RNA, and protein gradually increase between the globular and cotyledonary somatic embryos stages. DNA, RNA, and protein synthesis rates are in peak at the cotyledonary later stage, precotyledonary stage, and cotyledonary stage, respectively. It appears that more DNA, RNA, and protein are synthesized in the cotyledonary somatic embryo stage than in other stages. All these results indicate that an increased syntheses of DNA, RNA, and protein is associated with the differentiation of embryogenic cells and organogenesis in somatic embryos

  14. Chloroquine inhibits accessory cell presentation of soluble natural and synthetic protein antigens

    DEFF Research Database (Denmark)

    Buus, S; Werdelin, O

    1984-01-01

    We have studied the in vitro effect of the lysosomotrophic agent, chloroquine, on the presentation of soluble protein antigens by guinea pig accessory cells. Chloroquine inhibited the capacity of antigen-pulsed accessory cells to stimulate proliferation in appropriately primed T cells. The effect...... was time- and dose-dependent. A brief treatment solely of the accessory cells with the drug compromised their ability to stimulate primed T cells in a subsequent culture provided the accessory cells were treated with chloroquine before their exposure to the antigen. These results suggest that chloroquine...... acts on an early event in the antigen handling by accessory cells. Chloroquine is a well known inhibitor of lysosomal proteolysis, and it is likely that its effect on antigen presentation is caused by an inhibition of antigen degradation....

  15. In vitro study of proteins surface activity by tritium probe

    International Nuclear Information System (INIS)

    Chernysheva, M.G.; Badun, G.A.

    2010-01-01

    A new technique for in vitro studies of biomacromolecules interactions, their adsorption at aqueous/organic liquid interfaces and distribution in the bulk of liquid/liquid systems was developed. The method includes (1) tritium labeling of biomolecules by tritium thermal activation method and (2) scintillation phase step with organic phase, which can be concerned as a model of cellular membrane. Two globular proteins lysozyme and human serum albumin tested. We have determined the conditions of tritium labeling when labeled by-products can be easy separated by means of dialysis and size-exclusion chromatography. Scintillation phase experiments were conducted for three types of organic liquids. Thus, the influences of the nature of organic phase on proteins adsorption and its distribution in the bulk of aqueous/organic liquid system were determined. It was found that proteins possess high surface activity at aqueous/organic liquid interface. Furthermore, values of hydrophobicity of globular proteins were found by the experiment. (author)

  16. Coregulation of Soybean Vegetative Storage Protein Gene Expression by Methyl Jasmonate and Soluble Sugars 1

    Science.gov (United States)

    Mason, Hugh S.; DeWald, Daryll B.; Creelman, Robert A.; Mullet, John E.

    1992-01-01

    The soybean vegetative storage protein genes vspA and vspB are highly expressed in developing leaves, stems, flowers, and pods as compared with roots, seeds, and mature leaves and stems. In this paper, we report that physiological levels of methyl jasmonate (MeJA) and soluble sugars synergistically stimulate accumulation of vsp mRNAs. Treatment of excised mature soybean (Glycine max Merr. cv Williams) leaves with 0.2 molar sucrose and 10 micromolar MeJA caused a large accumulation of vsp mRNAs, whereas little accumulation occurred when these compounds were supplied separately. In soybean cell suspension cultures, the synergistic effect of sucrose and MeJA on the accumulation of vspB mRNA was maximal at 58 millimolar sucrose and was observed with fructose or glucose substituted for sucrose. In dark-grown soybean seedlings, the highest levels of vsp mRNAs occurred in the hypocotyl hook, which also contained high levels of MeJA and soluble sugars. Lower levels of vsp mRNAs, MeJA, and soluble sugars were found in the cotyledons, roots, and nongrowing regions of the stem. Wounding of mature soybean leaves induced a large accumulation of vsp mRNAs when wounded plants were incubated in the light. Wounded plants kept in the dark or illuminated plants sprayed with dichlorophenyldimethylurea, an inhibitor of photosynthetic electron transport, showed a greatly reduced accumulation of vsp mRNAs. The time courses for the accumulation of vsp mRNAs induced by wounding or sucrose/MeJA treatment were similar. These results strongly suggest that vsp expression is coregulated by endogenous levels of MeJA (or jasmonic acid) and soluble carbohydrate during normal vegetative development and in wounded leaves. ImagesFigure 1Figure 4Figure 5 PMID:16668757

  17. Gravitational microlensing by low-mass objects in the globular cluster M22.

    Science.gov (United States)

    Sahu, K C; Casertano, S; Livio, M; Gilliland, R L; Panagia, N; Albrow, M D; Potter, M

    2001-06-28

    Gravitational microlensing offers a means of determining directly the masses of objects ranging from planets to stars, provided that the distances and motions of the lenses and sources can be determined. A globular cluster observed against the dense stellar field of the Galactic bulge presents ideal conditions for such observations because the probability of lensing is high and the distances and kinematics of the lenses and sources are well constrained. The abundance of low-mass objects in a globular cluster is of particular interest, because it may be representative of the very early stages of star formation in the Universe, and therefore indicative of the amount of dark baryonic matter in such clusters. Here we report a microlensing event associated with the globular cluster M22. We determine the mass of the lens to be 0.13(+0.03)(-0.02) solar masses. We have also detected six events that are unresolved in time. If these are also microlensing events, they imply that a non-negligible fraction of the cluster mass resides in the form of free-floating planetary-mass objects.

  18. The Luminosity Functions of Old and Intermediate-Age Globular Clusters in NGC 3610

    OpenAIRE

    Whitmore, B. C.; Schweizer, F.; Kundu, A.; Miller, B. W.

    2002-01-01

    The WFPC2 Camera on board HST has been used to obtain high-resolution images of NGC 3610, a dynamically young elliptical galaxy. These observations supersede shorter, undithered HST observations where an intermediate-age population of globular clusters was first discovered. The new observations show the bimodal color distribution of globular clusters more clearly, with peaks at (V-I)o = 0.95 and 1.17. The luminosity function (LF) of the blue, metal-poor population of clusters in NGC 3610 turn...

  19. A tensegrity model for hydrogen bond networks in proteins

    Directory of Open Access Journals (Sweden)

    Robert P. Bywater

    2017-05-01

    Full Text Available Hydrogen-bonding networks in proteins considered as structural tensile elements are in balance separately from any other stabilising interactions that may be in operation. The hydrogen bond arrangement in the network is reminiscent of tensegrity structures in architecture and sculpture. Tensegrity has been discussed before in cells and tissues and in proteins. In contrast to previous work only hydrogen bonds are studied here. The other interactions within proteins are either much stronger − covalent bonds connecting the atoms in the molecular skeleton or weaker forces like the so-called hydrophobic interactions. It has been demonstrated that the latter operate independently from hydrogen bonds. Each category of interaction must, if the protein is to have a stable structure, balance out. The hypothesis here is that the entire hydrogen bond network is in balance without any compensating contributions from other types of interaction. For sidechain-sidechain, sidechain-backbone and backbone-backbone hydrogen bonds in proteins, tensegrity balance (“closure” is required over the entire length of the polypeptide chain that defines individually folding units in globular proteins (“domains” as well as within the repeating elements in fibrous proteins that consist of extended chain structures. There is no closure to be found in extended structures that do not have repeating elements. This suggests an explanation as to why globular domains, as well as the repeat units in fibrous proteins, have to have a defined number of residues. Apart from networks of sidechain-sidechain hydrogen bonds there are certain key points at which this closure is achieved in the sidechain-backbone hydrogen bonds and these are associated with demarcation points at the start or end of stretches of secondary structure. Together, these three categories of hydrogen bond achieve the closure that is necessary for the stability of globular protein domains as well as repeating

  20. A stochastic finite element model for the dynamics of globular macromolecules

    Science.gov (United States)

    Oliver, Robin C.; Read, Daniel J.; Harlen, Oliver G.; Harris, Sarah A.

    2013-04-01

    We describe a novel coarse-grained simulation method for modelling the dynamics of globular macromolecules, such as proteins. The macromolecule is treated as a continuum that is subject to thermal fluctuations. The model includes a non-linear treatment of elasticity and viscosity with thermal noise that is solved using finite element analysis. We have validated the method by demonstrating that the model provides average kinetic and potential energies that are in agreement with the classical equipartition theorem and that the nodal velocities have the correct Gaussian distribution. In addition, we have performed Fourier analysis on the simulation trajectories obtained for a series of linear beams to confirm that the correct average energies are present in the first two Fourier bending modes and that the probability distribution of the amplitudes of the first two Fourier modes match the theoretical results. We demonstrate spatial convergence of the model by showing that the anisotropy of the inertia tensor for a cubic mesh converges as a function of the mesh resolution. We have then used the new modelling method to simulate the thermal fluctuations of a representative protein over 500 ns timescales. Using reasonable parameters for the material properties, we have demonstrated that the overall deformation of the biomolecule is consistent with the results obtained for proteins in general from atomistic molecular dynamics simulations.

  1. Trazando la materia oscura con cúmulos globulares

    Science.gov (United States)

    Forte, J. C.

    Se describe la estrategia adoptada para mapear la distribución de materia oscura y bariónica en galaxias elípticas cuyos cúmulos globulares están siendo observados con los telescopios VLT y Gemini. Se ejemplifican los resultados con los datos obtenidos en el cúmulo de Fornax.

  2. Dating the Tidal Disruption of Globular Clusters with GAIA Data on Their Stellar Streams

    Science.gov (United States)

    Bose, Sownak; Ginsburg, Idan; Loeb, Abraham

    2018-05-01

    The Gaia mission promises to deliver precision astrometry at an unprecedented level, heralding a new era for discerning the kinematic and spatial coordinates of stars in our Galaxy. Here, we present a new technique for estimating the age of tidally disrupted globular cluster streams using the proper motions and parallaxes of tracer stars. We evolve the collisional dynamics of globular clusters within the evolving potential of a Milky Way-like halo extracted from a cosmological ΛCDM simulation and analyze the resultant streams as they would be observed by Gaia. The simulations sample a variety of globular cluster orbits, and account for stellar evolution and the gravitational influence of the disk of the Milky Way. We show that a characteristic timescale, obtained from the dispersion of the proper motions and parallaxes of stars within the stream, is a good indicator for the time elapsed since the stream has been freely expanding away due to the tidal disruption of the globular cluster. This timescale, in turn, places a lower limit on the age of the cluster. The age can be deduced from astrometry using a modest number of stars, with the error on this estimate depending on the proximity of the stream and the number of tracer stars used.

  3. On the importance of polar interactions for complexes containing intrinsically disordered proteins.

    Directory of Open Access Journals (Sweden)

    Eric T C Wong

    Full Text Available There is a growing recognition for the importance of proteins with large intrinsically disordered (ID segments in cell signaling and regulation. ID segments in these proteins often harbor regions that mediate molecular recognition. Coupled folding and binding of the recognition regions has been proposed to confer high specificity to interactions involving ID segments. However, researchers recently questioned the origin of the interaction specificity of ID proteins because of the overrepresentation of hydrophobic residues in their interaction interfaces. Here, we focused on the role of polar and charged residues in interactions mediated by ID segments. Making use of the extended nature of most ID segments when in complex with globular proteins, we first identified large numbers of complexes between globular proteins and ID segments by using radius-of-gyration-based selection criteria. Consistent with previous studies, we found the interfaces of these complexes to be enriched in hydrophobic residues, and that these residues contribute significantly to the stability of the interaction interface. However, our analyses also show that polar interactions play a larger role in these complexes than in structured protein complexes. Computational alanine scanning and salt-bridge analysis indicate that interfaces in ID complexes are highly complementary with respect to electrostatics, more so than interfaces of globular proteins. Follow-up calculations of the electrostatic contributions to the free energy of binding uncovered significantly stronger Coulombic interactions in complexes harbouring ID segments than in structured protein complexes. However, they are counter-balanced by even higher polar-desolvation penalties. We propose that polar interactions are a key contributing factor to the observed high specificity of ID segment-mediated interactions.

  4. Degradation of some representative polycyclic aromatic hydrocarbons by the water-soluble protein extracts from Zea mays L. cv PR32-B10.

    Science.gov (United States)

    Barone, Roberto; de Biasi, Margherita-Gabriella; Piccialli, Vincenzo; de Napoli, Lorenzo; Oliviero, Giorgia; Borbone, Nicola; Piccialli, Gennaro

    2016-10-01

    The ability of the water-soluble protein extracts from Zea mais L. cv. PR32-B10 to degrade some representative polycyclic aromatic hydrocarbons (PAHs), has been evaluated. Surface sterilized seeds of corn (Zea mais L. Pioneer cv. PR32-B10) were hydroponically cultivated in a growth chamber under no-stressful conditions. The water-soluble protein extracts isolated from maize tissues showed peroxidase, polyphenol oxidase and catalase activities. Incubation of the extracts with naphthalene, fluorene, phenanthrene and pyrene, led to formation of oxidized and/or degradation products. GC-MS and TLC monitoring of the processes showed that naphthalene, phenanthrene, fluorene and pyrene underwent 100%, 78%, 92% and 65% oxidative degradation, respectively, after 120 min. The chemical structure of the degradation products were determined by (1)H NMR and ESI-MS spectrometry. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Molecular sieving action of the cell membrane during gradual osmotic hemolysis

    Energy Technology Data Exchange (ETDEWEB)

    MacGregor, R.D. II

    1977-05-01

    Rat erythrocytes were hemolyzed by controlled gradual osmotic hemolysis to study cell morphology and hemoglobin loss from individual cells. Results suggest that each increase in the rate of loss of a protein from the cells during the initial phases of controlled gradual osmotic hemolysis is caused by the passage of a previously impermeable species across the stressed membrane. Similarly, during the final stages of controlled gradual osmotic hemolysis, each sharp decrease in the rate of loss of a protein corresponds to the termination of a molecular flow. A theoretical model is described that predicts the molecular sieving of soluble globular proteins across the stressed red cell membrane. Hydrophobic interactions occur between the soluble proteins and the lipid bilayer portion of the cell membrane. A spectrin network subdivides the bilayer into domains that restrict the insertion of large molecules into the membrane. Other membrane proteins affect soluble protein access to the membrane. Changes in the loss curves caused by incubation of red cells are discussed in terms of the model.

  6. Unique Features of Halophilic Proteins.

    Science.gov (United States)

    Arakawa, Tsutomu; Yamaguchi, Rui; Tokunaga, Hiroko; Tokunaga, Masao

    2017-01-01

    Proteins from moderate and extreme halophiles have unique characteristics. They are highly acidic and hydrophilic, similar to intrinsically disordered proteins. These characteristics make the halophilic proteins soluble in water and fold reversibly. In addition to reversible folding, the rate of refolding of halophilic proteins from denatured structure is generally slow, often taking several days, for example, for extremely halophilic proteins. This slow folding rate makes the halophilic proteins a novel model system for folding mechanism analysis. High solubility and reversible folding also make the halophilic proteins excellent fusion partners for soluble expression of recombinant proteins.

  7. TIME-SERIES PHOTOMETRY OF GLOBULAR CLUSTERS: M62 (NGC 6266), THE MOST RR LYRAE-RICH GLOBULAR CLUSTER IN THE GALAXY?

    International Nuclear Information System (INIS)

    Contreras, R.; Catelan, M.; Smith, H. A.; Kuehn, C. A.; Pritzl, B. J.; Borissova, J.

    2010-01-01

    We present new time-series CCD photometry, in the B and V bands, for the moderately metal-rich ([Fe/H] ≅ -1.3) Galactic globular cluster M62 (NGC 6266). The present data set is the largest obtained so far for this cluster and consists of 168 images per filter, obtained with the Warsaw 1.3 m telescope at the Las Campanas Observatory and the 1.3 m telescope of the Cerro Tololo Inter-American Observatory, in two separate runs over the time span of 3 months. The procedure adopted to detect the variable stars was the optimal image subtraction method (ISIS v2.2), as implemented by Alard. The photometry was performed using both ISIS and Stetson's DAOPHOT/ALLFRAME package. We have identified 245 variable stars in the cluster fields that have been analyzed so far, of which 179 are new discoveries. Of these variables, 133 are fundamental mode RR Lyrae stars (RRab), 76 are first overtone (RRc) pulsators, 4 are type II Cepheids, 25 are long-period variables (LPVs), 1 is an eclipsing binary, and 6 are not yet well classified. Such a large number of RR Lyrae stars places M62 among the top two most RR Lyrae-rich (in the sense of total number of RR Lyrae stars present) globular clusters known in the Galaxy, second only to M3 (NGC 5272) with a total of 230 known RR Lyrae stars. Since this study covers most but not all of the cluster area, it is not unlikely that M62 is in fact the most RR Lyrae-rich globular cluster in the Galaxy. In like vein, thanks to the time coverage of our data sets, we were also able to detect the largest sample of LPVs known so far in a Galactic globular cluster. We analyze a variety of Oosterhoff type indicators for the cluster, including mean periods, period distribution, Bailey diagrams, and Fourier decomposition parameters (as well as the physical parameters derived therefrom). All of these indicators clearly show that M62 is an Oosterhoff type I system. This is in good agreement with the moderately high metallicity of the cluster, in spite of its

  8. A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes

    International Nuclear Information System (INIS)

    Caffrey, Martin

    2015-01-01

    A comprehensive and up-to-date review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes is reported. Recent applications of the method for in situ serial crystallography at X-ray free-electron lasers and synchrotrons are described. The lipid cubic phase or in meso method is a robust approach for crystallizing membrane proteins for structure determination. The uptake of the method is such that it is experiencing what can only be described as explosive growth. This timely, comprehensive and up-to-date review introduces the reader to the practice of in meso crystallogenesis, to the associated challenges and to their solutions. A model of how crystallization comes about mechanistically is presented for a more rational approach to crystallization. The possible involvement of the lamellar and inverted hexagonal phases in crystallogenesis and the application of the method to water-soluble, monotopic and lipid-anchored proteins are addressed. How to set up trials manually and automatically with a robot is introduced with reference to open-access online videos that provide a practical guide to all aspects of the method. These range from protein reconstitution to crystal harvesting from the hosting mesophase, which is noted for its viscosity and stickiness. The sponge phase, as an alternative medium in which to perform crystallization, is described. The compatibility of the method with additive lipids, detergents, precipitant-screen components and materials carried along with the protein such as denaturants and reducing agents is considered. The powerful host and additive lipid-screening strategies are described along with how samples that have low protein concentration and cell-free expressed protein can be used. Assaying the protein reconstituted in the bilayer of the cubic phase for function is an important element of quality control and is detailed. Host lipid design for crystallization at low temperatures and for

  9. A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes

    Energy Technology Data Exchange (ETDEWEB)

    Caffrey, Martin, E-mail: martin.caffrey@tcd.ie [Trinity College Dublin, Dublin (Ireland)

    2015-01-01

    A comprehensive and up-to-date review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes is reported. Recent applications of the method for in situ serial crystallography at X-ray free-electron lasers and synchrotrons are described. The lipid cubic phase or in meso method is a robust approach for crystallizing membrane proteins for structure determination. The uptake of the method is such that it is experiencing what can only be described as explosive growth. This timely, comprehensive and up-to-date review introduces the reader to the practice of in meso crystallogenesis, to the associated challenges and to their solutions. A model of how crystallization comes about mechanistically is presented for a more rational approach to crystallization. The possible involvement of the lamellar and inverted hexagonal phases in crystallogenesis and the application of the method to water-soluble, monotopic and lipid-anchored proteins are addressed. How to set up trials manually and automatically with a robot is introduced with reference to open-access online videos that provide a practical guide to all aspects of the method. These range from protein reconstitution to crystal harvesting from the hosting mesophase, which is noted for its viscosity and stickiness. The sponge phase, as an alternative medium in which to perform crystallization, is described. The compatibility of the method with additive lipids, detergents, precipitant-screen components and materials carried along with the protein such as denaturants and reducing agents is considered. The powerful host and additive lipid-screening strategies are described along with how samples that have low protein concentration and cell-free expressed protein can be used. Assaying the protein reconstituted in the bilayer of the cubic phase for function is an important element of quality control and is detailed. Host lipid design for crystallization at low temperatures and for

  10. The nature of the globular- to fibrous-actin transition.

    Science.gov (United States)

    Oda, Toshiro; Iwasa, Mitsusada; Aihara, Tomoki; Maéda, Yuichiro; Narita, Akihiro

    2009-01-22

    Actin plays crucial parts in cell motility through a dynamic process driven by polymerization and depolymerization, that is, the globular (G) to fibrous (F) actin transition. Although our knowledge about the actin-based cellular functions and the molecules that regulate the G- to F-actin transition is growing, the structural aspects of the transition remain enigmatic. We created a model of F-actin using X-ray fibre diffraction intensities obtained from well oriented sols of rabbit skeletal muscle F-actin to 3.3 A in the radial direction and 5.6 A along the equator. Here we show that the G- to F-actin conformational transition is a simple relative rotation of the two major domains by about 20 degrees. As a result of the domain rotation, the actin molecule in the filament is flat. The flat form is essential for the formation of stable, helical F-actin. Our F-actin structure model provides the basis for understanding actin polymerization as well as its molecular interactions with actin-binding proteins.

  11. Close stellar encounters in globular clusters

    International Nuclear Information System (INIS)

    Bailyn, C.D.

    1989-01-01

    Stellar encounters are expected to produce a variety of interesting objects in the cores of globular clusters, either through the formation of binaries by tidal capture, or direct collisions. Here, I describe several attempts to observe the products of stellar encounters. In particular, the use of color maps has demonstrated the existence of a color gradient in the core of M15, which seems to be caused by a population of faint blue objects concentrated towards the cluster center. (author)

  12. Color Gradients Within Globular Clusters: Restricted Numerical Simulation

    Directory of Open Access Journals (Sweden)

    Young-Jong Sohn

    1997-06-01

    Full Text Available The results of a restricted numerical simulation for the color gradients within globular clusters have been presented. The standard luminosity function of M3 and Salpeter's initial mass functions were used to generate model clusters as a fundamental population. Color gradients with the sample clusters for both King and power law cusp models of surface brightness distributions are discussed in the case of using the standard luminosity function. The dependence of color gradients on several parameters for the simulations with Salpeter's initial mass functions, such as slope of initial mass functions, cluster ages, metallicities, concentration parameters of King model, and slopes of power law, are also discussed. No significant radial color gradients are shown to the sample clusters which are regenerated by a random number generation technique with various parameters in both of King and power law cusp models of surface brightness distributions. Dynamical mass segregation and stellar evolution of horizontal branch stars and blue stragglers should be included for the general case of model simulations to show the observed radial color gradients within globular clusters.

  13. Colon-specific delivery of a probiotic-derived soluble protein ameliorates intestinal inflammation in mice through an EGFR-dependent mechanism

    Science.gov (United States)

    Yan, Fang; Cao, Hanwei; Cover, Timothy L.; Washington, M. Kay; Shi, Yan; Liu, LinShu; Chaturvedi, Rupesh; Peek, Richard M.; Wilson, Keith T.; Polk, D. Brent

    2011-01-01

    Probiotic bacteria can potentially have beneficial effects on the clinical course of several intestinal disorders, but our understanding of probiotic action is limited. We have identified a probiotic bacteria–derived soluble protein, p40, from Lactobacillus rhamnosus GG (LGG), which prevents cytokine-induced apoptosis in intestinal epithelial cells. In the current study, we analyzed the mechanisms by which p40 regulates cellular responses in intestinal epithelial cells and p40’s effects on experimental colitis using mouse models. We show that the recombinant p40 protein activated EGFR, leading to Akt activation. Activation of EGFR by p40 was required for inhibition of cytokine-induced apoptosis in intestinal epithelial cells in vitro and ex vivo. Furthermore, we developed a pectin/zein hydrogel bead system to specifically deliver p40 to the mouse colon, which activated EGFR in colon epithelial cells. Administration of p40-containing beads reduced intestinal epithelial apoptosis and disruption of barrier function in the colon epithelium in an EGFR-dependent manner, thereby preventing and treating DSS-induced intestinal injury and acute colitis. Furthermore, p40 activation of EGFR was required for ameliorating colon epithelial cell apoptosis and chronic inflammation in oxazolone-induced colitis. These data define what we believe to be a previously unrecognized mechanism of probiotic-derived soluble proteins in protecting the intestine from injury and inflammation. PMID:21606592

  14. Identification of the hemoglobin scavenger receptor/CD163 as a natural soluble protein in plasma

    DEFF Research Database (Denmark)

    Møller, Holger Jon; Peterslund, Niels Anker; Graversen, Jonas Heilskov

    2002-01-01

    enabled identification of a soluble plasma form of HbSR (sHbSR) having an electrophoretic mobility equal to that of recombinant HbSR consisting of the extracellular domain (scavenger receptor cysteine-rich 1-9). A sandwich enzyme-linked immunosorbent assay was established and used to measure the s...... a level of sHbSR above the range of healthy persons. Patients with myelomonocytic leukemias and pneumonia/sepsis exhibited the highest levels (up to 67.3 mg/L). In conclusion, sHbSR is an abundant plasma protein potentially valuable in monitoring patients with infections and myelomonocytic leukemia....

  15. The structure of the Myo4p globular tail and its function in ASH1 mRNA localization.

    Science.gov (United States)

    Heuck, Alexander; Fetka, Ingrid; Brewer, Daniel N; Hüls, Daniela; Munson, Mary; Jansen, Ralf-Peter; Niessing, Dierk

    2010-05-03

    Type V myosin (MyoV)-dependent transport of cargo is an essential process in eukaryotes. Studies on yeast and vertebrate MyoV showed that their globular tails mediate binding to the cargo complexes. In Saccharomyces cerevisiae, the MyoV motor Myo4p interacts with She3p to localize asymmetric synthesis of HO 1 (ASH1) mRNA into the bud of dividing cells. A recent study showed that localization of GFP-MS2-tethered ASH1 particles does not require the Myo4p globular tail, challenging the supposed role of this domain. We assessed ASH1 mRNA and Myo4p distribution more directly and found that their localization is impaired in cells expressing globular tail-lacking Myo4p. In vitro studies further show that the globular tail together with a more N-terminal linker region is required for efficient She3p binding. We also determined the x-ray structure of the Myo4p globular tail and identify a conserved surface patch important for She3p binding. The structure shows pronounced similarities to membrane-tethering complexes and indicates that Myo4p may not undergo auto-inhibition of its motor domain.

  16. Extremely stable soluble high molecular mass multi-protein complex with DNase activity in human placental tissue.

    Directory of Open Access Journals (Sweden)

    Evgeniya E Burkova

    Full Text Available Human placenta is an organ which protects, feeds, and regulates the grooving of the embryo. Therefore, identification and characterization of placental components including proteins and their multi-protein complexes is an important step to understanding the placenta function. We have obtained and analyzed for the first time an extremely stable multi-protein complex (SPC, ∼ 1000 kDa from the soluble fraction of three human placentas. By gel filtration on Sepharose-4B, the SPC was well separated from other proteins of the placenta extract. Light scattering measurements and gel filtration showed that the SPC is stable in the presence of NaCl, MgCl2, acetonitrile, guanidinium chloride, and Triton in high concentrations, but dissociates efficiently in the presence of 8 M urea, 50 mM EDTA, and 0.5 M NaCl. Such a stable complex is unlikely to be a casual associate of different proteins. According to SDS-PAGE and MALDI mass spectrometry data, this complex contains many major glycosylated proteins with low and moderate molecular masses (MMs 4-14 kDa and several moderately abundant (79.3, 68.5, 52.8, and 27.2 kDa as well as minor proteins with higher MMs. The SPC treatment with dithiothreitol led to a disappearance of some protein bands and revealed proteins with lower MMs. The SPCs from three placentas efficiently hydrolyzed plasmid supercoiled DNA with comparable rates and possess at least two DNA-binding sites with different affinities for a 12-mer oligonucleotide. Progress in study of placental protein complexes can promote understanding of their biological functions.

  17. Effect of the combinations between pea proteins and soluble fibres on cholesterolaemia and cholesterol metabolism in rats.

    Science.gov (United States)

    Parolini, Cinzia; Manzini, Stefano; Busnelli, Marco; Rigamonti, Elena; Marchesi, Marta; Diani, Erika; Sirtori, Cesare R; Chiesa, Giulia

    2013-10-01

    Many functional foods and dietary supplements have been reported to be beneficial for the management of dyslipidaemia, one of the major risk factors for CVD. Soluble fibres and legume proteins are known to be a safe and practical approach for cholesterol reduction. The present study aimed at investigating the hypocholesterolaemic effect of the combinations of these bioactive vegetable ingredients and their possible effects on the expression of genes regulating cholesterol homeostasis. A total of six groups of twelve rats each were fed, for 28 d, Nath's hypercholesterolaemic diets, differing in protein and fibre sources, being, respectively, casein and cellulose (control), pea proteins and cellulose (pea), casein and oat fibres (oat), casein and apple pectin (pectin), pea proteins and oat fibres (pea+oat) and pea proteins and apple pectin (pea+pectin). Administration of each vegetable-containing diet was associated with lower total cholesterol concentrations compared with the control. The combinations (pea+oat and pea+pectin) were more efficacious than fibres alone in modulating cholesterolaemia ( - 53 and - 54%, respectively, at 28 d; Ppea proteins, a lower hepatic cholesterol content (Ppea proteins and oat fibres or apple pectin are extremely effective in lowering plasma cholesterol concentrations in rats and affect cellular cholesterol homeostasis by up-regulating genes involved in hepatic cholesterol turnover.

  18. Formation of soluble amyloid oligomers and amyloid fibrils by the multifunctional protein vitronectin

    Directory of Open Access Journals (Sweden)

    Langen Ralf

    2008-10-01

    Full Text Available Abstract Background The multifunctional protein vitronectin is present within the deposits associated with Alzheimer disease (AD, age-related macular degeneration (AMD, atherosclerosis, systemic amyloidoses, and glomerulonephritis. The extent to which vitronectin contributes to amyloid formation within these plaques, which contain misfolded, amyloidogenic proteins, and the role of vitronectin in the pathophysiology of the aforementioned diseases is currently unknown. The investigation of vitronectin aggregation is significant since the formation of oligomeric and fibrillar structures are common features of amyloid proteins. Results We observed vitronectin immunoreactivity in senile plaques of AD brain, which exhibited overlap with the amyloid fibril-specific OC antibody, suggesting that vitronectin is deposited at sites of amyloid formation. Of particular interest is the growing body of evidence indicating that soluble nonfibrillar oligomers may be responsible for the development and progression of amyloid diseases. In this study we demonstrate that both plasma-purified and recombinant human vitronectin readily form spherical oligomers and typical amyloid fibrils. Vitronectin oligomers are toxic to cultured neuroblastoma and retinal pigment epithelium (RPE cells, possibly via a membrane-dependent mechanism, as they cause leakage of synthetic vesicles. Oligomer toxicity was attenuated in RPE cells by the anti-oligomer A11 antibody. Vitronectin fibrils contain a C-terminal protease-resistant fragment, which may approximate the core region of residues essential to amyloid formation. Conclusion These data reveal the propensity of vitronectin to behave as an amyloid protein and put forth the possibilities that accumulation of misfolded vitronectin may contribute to aggregate formation seen in age-related amyloid diseases.

  19. Design and performance of an ultraviolet resonance Raman spectrometer for proteins and nucleic acids.

    Science.gov (United States)

    Russell, M P; Vohník, S; Thomas, G J

    1995-04-01

    We describe an ultraviolet resonance Raman (UVRR) spectrometer appropriate for structural studies of biological macromolecules and their assemblies. Instrument design includes the following features: a continuous wave, intracavity doubled, ultraviolet laser source for excitation of the Raman spectrum; a rotating cell (or jet source) for presentation of the sample to the laser beam; a Cassegrain optic with f/1.0 aperture for collection of the Raman scattering; a quartz prism dispersing element for rejection of stray light and Rayleigh scattering; a 0.75-m single grating monochromator for dispersion of the Raman scattering; and a liquid-nitrogen-cooled, charge-coupled device for detection of the Raman photons. The performance of this instrument, assessed on the basis of the observed signal-to-noise ratios, the apparent resolution of closely spaced spectral bands, and the wide spectrometer bandpass of 2200 cm-1, is believed superior to previously described UVRR spectrometers of similar design. Performance characteristics of the instrument are demonstrated in UVRR spectra obtained from standard solvents, p-ethylphenol, which serves as a model for the tyrosine side chain, the DNA nucleotide deoxyguanosine-5'-monophosphate, and the human tumor necrosis factor binding protein, which is considered representative of soluble globular proteins.

  20. The fragmentation of proto-globular clusters. I. Thermal instabilities

    International Nuclear Information System (INIS)

    Murray, S.D.; Lin, D.N.C.

    1989-01-01

    The metal abundances among the stars within a typical globular cluster are remarkably homogeneous. This indicates that star formation in these systems was a globally coordinated event which occurred over a time span less than or comparable to the collapse time scale of the cluster. This issue is addressed by assuming that the fragmentation of a proto-globular cluster cloud proceeded in two steps. In the first step, thermal instability led to the rapid growth of initial fluctuations. This led to a large contrast in the dynamical time scales between the perturbations and the parent cloud, and the perturbations then underwent gravitational instabilities on short time scales. This process is modeled using one-dimensional hydrodynamic simulations of clouds both with and without external heat sources and self-gravity. The models include the effects of a non-equilibrium H2 abundance. The results indicate that fragmentation can occur on time scales significantly less than the dynamical time scale of the parent cloud. 21 refs

  1. Globular hepatic amyloid is highly sensitive and specific for LECT2 amyloidosis.

    Science.gov (United States)

    Chandan, Vishal S; Shah, Sejal S; Lam-Himlin, Dora M; Petris, Giovanni De; Mereuta, Oana M; Dogan, Ahmet; Torbenson, Michael S; Wu, Tsung-Teh

    2015-04-01

    Globular hepatic amyloid (GHA) is rare, and its clinical significance remains unclear. Recently, leukocyte chemotactic factor-associated amyloidosis (ALECT2) has been reported to involve the liver, showing a globular pattern. We reviewed 70 consecutive cases of hepatic amyloidosis to determine the prevalence and morphology of hepatic amyloid subtypes, especially ALECT2 and its association with GHA. Each case was reviewed for amyloid subtype (immunohistochemistry and/or mass spectrometry), its pattern (linear or globular), and distribution (vascular, perisinusoidal, or stromal). In addition, 24 cases of confirmed hepatic ALECT2 on mass spectrometry from our consultation files were also reviewed. LECT2 immunostaining was performed in 49 cases. Of the 70 cases, immunoglobulin light chain (AL) type was most common with 41 cases (59%), followed by transthyretin (ATTR) 15 cases (22%), 3 cases each of fibrinogen A (AFib) (4%), serum amyloid A (AA) (4%), and ALECT2 (4%), 2 cases of apolipoproteins (AApoA1) (3%), and 3 cases (4%) were unclassified. Three of our 70 cases (4%), with ALECT2, and all 24 cases (100%) of mass spectrometry-confirmed hepatic ALECT2 showed only GHA deposits in the hepatic sinusoids and portal tracts. Three (4%) other cases of AL type showed a focal globular pattern admixed with prominent linear amyloid. None of the other amyloid subtypes showed GHA. LECT2 immunostain was positive in all 27 cases (100%) of ALECT2 and negative in the other 22 non-ALECT2 cases (100%) (14 AL, 5 ATTR, 1 AA, 1 AFib, 1 AApoA1). Pure GHA is uncommon (4%) but is highly specific for ALECT2, and LECT2 immunostain is helpful in confirming this amyloid type.

  2. The binding affinity of a soluble TCR-Fc fusion protein is significantly improved by crosslinkage with an anti-C{beta} antibody

    Energy Technology Data Exchange (ETDEWEB)

    Ozawa, Tatsuhiko; Horii, Masae; Kobayashi, Eiji [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan); Jin, Aishun [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan); Department of Immunology, College of Basic Medical Sciences, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081 (China); Kishi, Hiroyuki, E-mail: immkishi@med.u-toyama.ac.jp [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan); Muraguchi, Atsushi [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan)

    2012-06-01

    Highlights: Black-Right-Pointing-Pointer A novel soluble TCR composed of TCR V and C regions with Ig Fc region is generated. Black-Right-Pointing-Pointer TCR-Fc protein immobilized by an anti-C{beta} antibody bound to a p/MHC tetramer. Black-Right-Pointing-Pointer Binding affinity of TCR-Fc was markedly increased by binding with anti-C{beta} antibody. -- Abstract: The identification and cloning of tumor antigen-specific T cell receptors (TCRs) and the production of the soluble form of the TCR (sTCR) contributed to the development of diagnostic and therapeutic tools for cancer. Recently, several groups have reported the development of technologies for the production of sTCRs. The native sTCR has a very low binding affinity for the antigenic peptide/MHC (p/MHC) complex. In this study, we established a technology to produce high affinity, functional sTCRs. We generated a novel sTCR-Fc fusion protein composed of the TCR V and C regions of the TCR linked to the immunoglobulin (Ig) Fc region. A Western blot analysis revealed that the molecular weight of the fusion protein was approximately 60 kDa under reducing conditions and approximately 100-200 kDa under non-reducing conditions. ELISAs using various antibodies showed that the structure of each domain of the TCR-Fc protein was intact. The TCR-Fc protein immobilized by an anti-C{beta} antibody effectively bound to a p/MHC tetramer. An SPR analysis showed that the TCR-Fc protein had a low binding affinity (KD; 1.1 Multiplication-Sign 10{sup -5} M) to the p/MHC monomer. Interestingly, when the TCR-Fc protein was pre-incubated with an anti-C{beta} antibody, its binding affinity for p/MHC increased by 5-fold (2.2 Multiplication-Sign 10{sup -6} M). We demonstrated a novel method for constructing a functional soluble TCR using the Ig Fc region and showed that the binding affinity of the functional sTCR-Fc was markedly increased by an anti-C{beta} antibody, which is probably due to the stabilization of the V

  3. Globular Cluster Variable Stars—Atlas and Coordinate Improvement using AAVSOnet Telescopes (Abstract)

    Science.gov (United States)

    Welch, D.; Henden, A.; Bell, T.; Suen, C.; Fare, I.; Sills, A.

    2015-12-01

    (Abstract only) The variable stars of globular clusters have played and continue to play a significant role in our understanding of certain classes of variable stars. Since all stars associated with a cluster have the same age, metallicity, distance and usually very similar (if not identical reddenings), such variables can produce uniquely powerful constraints on where certain types of pulsation behaviors are excited. Advanced amateur astronomers are increasingly well-positioned to provide long-term CCD monitoring of globular cluster variable star but are hampered by a long history of poor or inaccessible finder charts and coordinates. Many of variable-rich clusters have published photographic finder charts taken in relatively poor seeing with blue-sensitive photographic plates. While useful signal-to-noise ratios are relatively straightforward to achieve for RR Lyrae, Type 2 Cepheids, and red giant variables, correct identification remains a difficult issue—particularly when images are taken at V or longer wavelengths. We describe the project and report its progress using the OC61, TMO61, and SRO telescopes of AAVSOnet after the first year of image acquisition and demonstrate several of the data products being developed for globular cluster variables.

  4. Parâmetros astrofísicos de estrelas gigantes do aglomerado globular 47 Tucanae

    Science.gov (United States)

    Alves-Brito, A.; Barbuy, B.

    2003-08-01

    Os aglomerados globulares são considerados laboratórios astrofísicos para a verificação da teoria de evolução estelar, bem como a trajetória químio-dinâmica das galáxias hospedeiras. Em particular, 47 Tucanae (NGC 104) configura-se como um dos mais extensivamente estudados aglomerados globulares da Galáxia devido a relativa proximidade ao Sol (R¤ = 4.5 kpc) e alta latitute galáctica (b = -44°,89). Neste trabalho, apresentamos a velocidade radial heliocêntrica e os parâmetros atmosféricos (Teff, logg, [Fe/H]) de 5 estrelas gigantes do aglomerado globular 47 Tucanae. Os espectros foram obtidos pelo espectrógrafo UVES (Ultaviolet Visual Echelle Spectrograph) de alta resolução (R = 60000) e alta razão sinal-ruído (S/N > 200), acoplado ao telescópio de 8,2m Kueyen do VLT (Very Large Telescope). Nós encontramos = -22,43 +/- 3,97 km/s, [Fe/H] ~ -0.7, 1,2 estrelas cobrem um intervalo de magnitude 12,2 < V < 14,2. Os parâmetros atmosféricos são fundamentais para a construção de espectros sintéticos de outros aglomerados globulares ricos em metais. Trabalho financiado pela FAPESP e pelo CNPq.

  5. Effect of soluble calcium and lactose on limiting flux and serum protein removal during skim milk microfiltration.

    Science.gov (United States)

    Adams, Michael C; Hurt, Emily E; Barbano, David M

    2015-11-01

    The tendency of calcium to promote microfiltration (MF) membrane fouling is well documented, but the role of lactose has not been studied. Milk protein concentrate that is 85% protein on a dry basis (MPC85) contains less calcium and lactose than skim milk. Our objectives were to determine the effects of skim milk soluble calcium and lactose concentrations on the limiting fluxes (LF) and serum protein (SP) removal factors of 0.1-µm ceramic graded permeability membranes. The MF was fed with 3 different milks: skim milk, liquid MPC85 that had been standardized to the protein content of skim milk with reverse osmosis water (MPC), and liquid MPC85 that had been standardized to the protein and lactose contents of skim milk with reverse osmosis water and lactose monohydrate (MPC+L). Retentate and permeate were continuously recycled to the feed tank. The LF for each feed was determined by increasing flux once per hour from 55 kg·m(-2)·h(-1) until flux did not increase with increasing transmembrane pressure. Temperature, pressure drop across the membrane length, and protein concentration in the retentate recirculation loop were maintained at 50°C, 220 kPa, and 8.77 ± 0.2%, respectively. Experiments were replicated 3 times and the Proc GLM procedure of SAS was used for statistical analysis. An increase in LF between skim milk (91 kg·m(-2)·h(-1)) and MPC+L (124 kg·m(-2)·h(-1)) was associated with a reduction in soluble calcium. The LF of MPC+L was lower than the LF of MPC (137 kg·m(-2)·h(-1)) due to the higher viscosity contributed by lactose. Permeates produced from the MPC and MPC+L contained more protein than the skim milk permeate due to the transfer of caseins from the micelles into the reduced-calcium sera of the MPC and MPC+L. A SP removal factor was calculated by dividing true protein in the permeate by SP in the permeate portion of the feed to describe the ease of SP passage through the membrane. No differences in SP removal factors were detected among the

  6. Conformational energy calculations on polypeptides and proteins: use of a statistical mechanical procedure for evaluating structure and properties.

    Science.gov (United States)

    Scheraga, H A; Paine, G H

    1986-01-01

    We are using a variety of theoretical and computational techniques to study protein structure, protein folding, and higher-order structures. Our earlier work involved treatments of liquid water and aqueous solutions of nonpolar and polar solutes, computations of the stabilities of the fundamental structures of proteins and their packing arrangements, conformations of small cyclic and open-chain peptides, structures of fibrous proteins (collagen), structures of homologous globular proteins, introduction of special procedures as constraints during energy minimization of globular proteins, and structures of enzyme-substrate complexes. Recently, we presented a new methodology for predicting polypeptide structure (described here); the method is based on the calculation of the probable and average conformation of a polypeptide chain by the application of equilibrium statistical mechanics in conjunction with an adaptive, importance sampling Monte Carlo algorithm. As a test, it was applied to Met-enkephalin.

  7. My 65 years in protein chemistry.

    Science.gov (United States)

    Scheraga, Harold A

    2015-05-01

    This is a tour of a physical chemist through 65 years of protein chemistry from the time when emphasis was placed on the determination of the size and shape of the protein molecule as a colloidal particle, with an early breakthrough by James Sumner, followed by Linus Pauling and Fred Sanger, that a protein was a real molecule, albeit a macromolecule. It deals with the recognition of the nature and importance of hydrogen bonds and hydrophobic interactions in determining the structure, properties, and biological function of proteins until the present acquisition of an understanding of the structure, thermodynamics, and folding pathways from a linear array of amino acids to a biological entity. Along the way, with a combination of experiment and theoretical interpretation, a mechanism was elucidated for the thrombin-induced conversion of fibrinogen to a fibrin blood clot and for the oxidative-folding pathways of ribonuclease A. Before the atomic structure of a protein molecule was determined by x-ray diffraction or nuclear magnetic resonance spectroscopy, experimental studies of the fundamental interactions underlying protein structure led to several distance constraints which motivated the theoretical approach to determine protein structure, and culminated in the Empirical Conformational Energy Program for Peptides (ECEPP), an all-atom force field, with which the structures of fibrous collagen-like proteins and the 46-residue globular staphylococcal protein A were determined. To undertake the study of larger globular proteins, a physics-based coarse-grained UNited-RESidue (UNRES) force field was developed, and applied to the protein-folding problem in terms of structure, thermodynamics, dynamics, and folding pathways. Initially, single-chain and, ultimately, multiple-chain proteins were examined, and the methodology was extended to protein-protein interactions and to nucleic acids and to protein-nucleic acid interactions. The ultimate results led to an understanding

  8. Evidence from stellar abundances for a large age difference between two globular clusters

    International Nuclear Information System (INIS)

    Dickens, R.J.; Croke, B.F.W.; Cannon, R.D.; Bell, R.A.

    1991-01-01

    The globular clusters NGC288 and NGC362 are central to recent claims of large age differences (∼3 Gyr) between globular clusters associated with our Galaxy. According to standard models for the formation of the Galaxy, the system of globular clusters formed during the dynamical collapse of the protogalactic cloud, a process which should have lasted no more than 1 Gyr. But the claimed age differences are derived from stellar evolution models using assumed CNO abundances, and uncertainty in the actual CNO abundances of about a factor of three could account for an apparent 2-Gyr age difference. We have accurately measured abundances in red giants in NGC288 and NGC362, and find that the Fe abundance and the sum of the C, N and O abundances are essentially the same in every star studied. By eliminating compositional differences and thus confirming the reality of the age difference, these results imply a cluster formation period that is hard to reconcile with the standard collapse model. (author)

  9. The endothelial protein C receptor rs867186-GG genotype is associated with increased soluble EPCR and could mediate protection against severe malaria

    DEFF Research Database (Denmark)

    Shabani, Estela; Opoka, Robert O; Bangirana, Paul

    2016-01-01

    The endothelial protein C receptor (EPCR) appears to play an important role in Plasmodium falciparum endothelial cell binding in severe malaria (SM). Despite consistent findings of elevated soluble EPCR (sEPCR) in other infectious diseases, field studies to date have provided conflicting data abo...

  10. The SLUGGS survey: globular cluster stellar population trends from weak absorption lines in stacked spectra

    Science.gov (United States)

    Usher, Christopher; Forbes, Duncan A.; Brodie, Jean P.; Romanowsky, Aaron J.; Strader, Jay; Conroy, Charlie; Foster, Caroline; Pastorello, Nicola; Pota, Vincenzo; Arnold, Jacob A.

    2015-01-01

    As part of the SAGES Legacy Unifying Globulars and GalaxieS (SLUGGS) survey, we stack 1137 Keck DEIMOS (Deep Imaging Multi-Object Spectrograph) spectra of globular clusters from 10 galaxies to study their stellar populations in detail. The stacked spectra have median signal-to-noise ratios of ˜90 Å-1. Besides the calcium triplet, we study weaker sodium, magnesium, titanium and iron lines as well as the Hα and higher order Paschen hydrogen lines. In general, the stacked spectra are consistent with old ages and a Milky Way-like initial mass function. However, we see different metal line index strengths at fixed colour and magnitude, and differences in the calcium triplet-colour relation from galaxy to galaxy. We interpret this as strong evidence for variations in the globular cluster colour-metallicity relation between galaxies. Two possible explanations for the colour-metallicity relation variations are that the average ages of globular clusters vary from galaxy to galaxy or that the average abundances of light elements (i.e. He, C, N and O) differ between galaxies. Stacking spectra by magnitude, we see that the colours become redder and metal line indices stronger with brighter magnitudes. These trends are consistent with the previously reported `blue tilts' being mass-metallicity relations.

  11. Contribution to the study of proteins and peptides structure by hydrogen isotopic exchange

    International Nuclear Information System (INIS)

    Nabedryk-Viala, Eliane.

    1978-01-01

    Development of hydrogen exchange measurement methods to study the structure and the molecular interaction of globular protein molecules in aqueous solution (ribonuclease A, cytochrome c, coupling factors of chloroplasts), in peptide hormones in trifluoroethanol solution (angiotensin II, corticotropin) and in proteins of membranes (rhodopsin) [fr

  12. Blockade of CD26-mediated T cell costimulation with soluble caveolin-1-Ig fusion protein induces anergy in CD4{sup +}T cells

    Energy Technology Data Exchange (ETDEWEB)

    Ohnuma, Kei [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Division of Clinical Immunology, The Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Uchiyama, Masahiko [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Computational Intelligence and System Science, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8503 (Japan); Hatano, Ryo; Takasawa, Wataru; Endo, Yuko [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Dang, Nam H. [Department of Hematologic Malignancies, Nevada Cancer Institute, Las Vegas, NV 89135 (United States); Morimoto, Chikao, E-mail: morimoto@ims.u-tokyo.ac.jp [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Division of Clinical Immunology, The Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan)

    2009-08-21

    CD26 binds to caveolin-1 in antigen-presenting cells (APC), and that ligation of CD26 by caveolin-1 induces T cell proliferation in a TCR/CD3-dependent manner. We report herein the effects of CD26-caveolin-1 costimulatory blockade by fusion protein caveolin-1-Ig (Cav-Ig). Soluble Cav-Ig inhibits T cell proliferation and cytokine production in response to recall antigen, or allogeneic APC. Our data hence suggest that blocking of CD26-associated signaling by soluble Cav-Ig may be an effective approach as immunosuppressive therapy.

  13. Mapping Soluble Guanylyl Cyclase and Protein Disulfide Isomerase Regions of Interaction.

    Directory of Open Access Journals (Sweden)

    Erin J Heckler

    Full Text Available Soluble guanylyl cyclase (sGC is a heterodimeric nitric oxide (NO receptor that produces cyclic GMP. This signaling mechanism is a key component in the cardiovascular system. NO binds to heme in the β subunit and stimulates the catalytic conversion of GTP to cGMP several hundred fold. Several endogenous factors have been identified that modulate sGC function in vitro and in vivo. In previous work, we determined that protein disulfide isomerase (PDI interacts with sGC in a redox-dependent manner in vitro and that PDI inhibited NO-stimulated activity in cells. To our knowledge, this was the first report of a physical interaction between sGC and a thiol-redox protein. To characterize this interaction between sGC and PDI, we first identified peptide linkages between sGC and PDI, using a lysine cross-linking reagent and recently developed mass spectrometry analysis. Together with Flag-immunoprecipitation using sGC domain deletions, wild-type (WT and mutated PDI, regions of sGC involved in this interaction were identified. The observed data were further explored with computational modeling to gain insight into the interaction mechanism between sGC and oxidized PDI. Our results indicate that PDI interacts preferentially with the catalytic domain of sGC, thus providing a mechanism for PDI inhibition of sGC. A model in which PDI interacts with either the α or the β catalytic domain is proposed.

  14. Structure and Dynamics of the Globular Cluster Palomar 13

    Science.gov (United States)

    Bradford, J. D.; Geha, M.; Muñoz, R. R.; Santana, F. A.; Simon, J. D.; Côté, P.; Stetson, P. B.; Kirby, E.; Djorgovski, S. G.

    2011-12-01

    We present Keck/DEIMOS spectroscopy and Canada-France-Hawaii Telescope/MegaCam photometry for the Milky Way globular cluster Palomar 13. We triple the number of spectroscopically confirmed members, including many repeat velocity measurements. Palomar 13 is the only known globular cluster with possible evidence for dark matter, based on a Keck/High Resolution Echelle Spectrometer 21 star velocity dispersion of σ = 2.2 ± 0.4 km s-1. We reproduce this measurement, but demonstrate that it is inflated by unresolved binary stars. For our sample of 61 stars, the velocity dispersion is σ = 0.7+0.6 -0.5 km s-1. Combining our DEIMOS data with literature values, our final velocity dispersion is σ = 0.4+0.4 -0.3 km s-1. We determine a spectroscopic metallicity of [Fe/H] = -1.6 ± 0.1 dex, placing a 1σ upper limit of σ[Fe/H] ~ 0.2 dex on any internal metallicity spread. We determine Palomar 13's total luminosity to be MV = -2.8 ± 0.4, making it among the least luminous known globular clusters. The photometric isophotes are regular out to the half-light radius and mildly irregular outside this radius. The outer surface brightness profile slope is shallower than typical globular clusters (Σvpropr η, η = -2.8 ± 0.3). Thus at large radius, tidal debris is likely affecting the appearance of Palomar 13. Combining our luminosity with the intrinsic velocity dispersion, we find a dynamical mass of M 1/2 = 1.3+2: 7 -1.3 × 103 M ⊙ and a mass-to-light ratio of M/LV = 2.4+5.0 -2.4 M ⊙/L ⊙. Within our measurement errors, the mass-to-light ratio agrees with the theoretical predictions for a single stellar population. We conclude that, while there is some evidence for tidal stripping at large radius, the dynamical mass of Palomar 13 is consistent with its stellar mass and neither significant dark matter, nor extreme tidal heating, is required to explain the cluster dynamics. The data presented herein were obtained at the W. M. Keck Observatory, which is operated as a

  15. Template mediated protein self-assembly as a valuable tool in regenerative therapy.

    Science.gov (United States)

    Kundu, B; Eltohamy, M; Yadavalli, V K; Reis, R L; Kim, H W

    2018-04-11

    The assembly of natural proteinaceous biopolymers into macro-scale architectures is of great importance in synthetic biology, soft-material science and regenerative therapy. The self-assembly of protein tends to be limited due to anisotropic interactions among protein molecules, poor solubility and stability. Here, we introduce a unique platform to self-immobilize diverse proteins (fibrous and globular, positively and negatively charged, low and high molecular weight) using silicon surfaces with pendant -NH 2 groups via a facile one step diffusion limited aggregation (DLA) method. All the experimental proteins (type I collagen, bovine serum albumin and cytochrome C) self-assemble into seaweed-like branched dendritic architectures via classical DLA in the absence of any electrolytes. The notable differences in branching architectures are due to dissimilarities in protein colloidal sub-units, which is typical for each protein type, along with the heterogeneous distribution of surface -NH 2 groups. Fractal analysis of assembled structures is used to explain the underlying route of fractal deposition; which concludes how proteins with different functionality can yield similar assembly. Further, the nano-micro-structured surfaces can be used to provide functional topographical cues to study cellular responses, as demonstrated using rat bone marrow stem cells. The results indicate that the immobilization of proteins via DLA does not affect functionality, instead serving as topographical cues to guide cell morphology. This indicates a promising design strategy at the tissue-material interface and is anticipated to guide future surface modifications. A cost-effective standard templating strategy is therefore proposed for fundamental and applied particle aggregation studies, which can be used at multiple length scales for biomaterial design and surface reformation.

  16. The Functional Quality of Soluble Recombinant Polypeptides Produced in Escherichia coli Is Defined by a Wide Conformational Spectrum▿

    Science.gov (United States)

    Martínez-Alonso, Mónica; González-Montalbán, Nuria; García-Fruitós, Elena; Villaverde, Antonio

    2008-01-01

    We have observed that a soluble recombinant green fluorescent protein produced in Escherichia coli occurs in a wide conformational spectrum. This results in differently fluorescent protein fractions in which morphologically diverse soluble aggregates abound. Therefore, the functional quality of soluble versions of aggregation-prone recombinant proteins is defined statistically rather than by the prevalence of a canonical native structure. PMID:18836021

  17. The SUMO project I. A survey of multiple populations in globular clusters

    Science.gov (United States)

    Monelli, M.; Milone, A. P.; Stetson, P. B.; Marino, A. F.; Cassisi, S.; del Pino Molina, A.; Salaris, M.; Aparicio, A.; Asplund, M.; Grundahl, F.; Piotto, G.; Weiss, A.; Carrera, R.; Cebrián, M.; Murabito, S.; Pietrinferni, A.; Sbordone, L.

    2013-05-01

    We present a general overview and the first results of the SUMO project (a SUrvey of Multiple pOpulations in Globular Clusters). The objective of this survey is the study of multiple stellar populations in the largest sample of globular clusters homogeneously analysed to date. To this aim we obtained high signal-to-noise (S/N > 50) photometry for main sequence stars with mass down to ˜0.5 M⊙ in a large sample of clusters using both archival and proprietary U, B, V and I data from ground-based telescopes. In this paper, we focus on the occurrence of multiple stellar populations in 23 clusters. We define a new photometric index, cU, B, I = (U - B) - (B - I), which turns out to be very effective for identifying multiple sequences along the red giant branch (RGB). We found that in the V-cU, B, I diagram all clusters presented in this paper show broadened or multimodal RGBs, with the presence of two or more components. We found a direct connection with the chemical properties of different sequences, which display different abundances of light elements (O, Na, C, N and Al). The cU, B, I index is also a powerful tool for identifying distinct sequences of stars along the horizontal branch and, for the first time in the case of NGC 104 (47 Tuc), along the asymptotic giant branch. Our results demonstrate that (i) the presence of more than two stellar populations is a common feature amongst globular clusters, as already highlighted in previous work; (ii) multiple sequences with different chemical contents can be easily identified by using standard Johnson photometry obtained with ground-based facilities; (iii) in the study of globular cluster multiple stellar populations the cU, B, I index is an alternative to spectroscopy, and has the advantage of larger statistics.

  18. Structural features that predict real-value fluctuations of globular proteins.

    Science.gov (United States)

    Jamroz, Michal; Kolinski, Andrzej; Kihara, Daisuke

    2012-05-01

    It is crucial to consider dynamics for understanding the biological function of proteins. We used a large number of molecular dynamics (MD) trajectories of nonhomologous proteins as references and examined static structural features of proteins that are most relevant to fluctuations. We examined correlation of individual structural features with fluctuations and further investigated effective combinations of features for predicting the real value of residue fluctuations using the support vector regression (SVR). It was found that some structural features have higher correlation than crystallographic B-factors with fluctuations observed in MD trajectories. Moreover, SVR that uses combinations of static structural features showed accurate prediction of fluctuations with an average Pearson's correlation coefficient of 0.669 and a root mean square error of 1.04 Å. This correlation coefficient is higher than the one observed in predictions by the Gaussian network model (GNM). An advantage of the developed method over the GNMs is that the former predicts the real value of fluctuation. The results help improve our understanding of relationships between protein structure and fluctuation. Furthermore, the developed method provides a convienient practial way to predict fluctuations of proteins using easily computed static structural features of proteins. Copyright © 2012 Wiley Periodicals, Inc.

  19. Globular adiponectin ameliorates metabolic insulin resistance via AMPK-mediated restoration of microvascular insulin responses

    Science.gov (United States)

    Zhao, Lina; Fu, Zhuo; Wu, Jing; Aylor, Kevin W; Barrett, Eugene J; Cao, Wenhong; Liu, Zhenqi

    2015-01-01

    Abstract Hypoadiponectinaemia is closely associated with endothelial dysfunction and insulin resistance, and microvasculature plays a critical role in the regulation of insulin action in muscle. Here we tested whether adiponectin replenishment could improve metabolic insulin sensitivity in male rats fed a high-fat diet (HFD) via the modulation of microvascular insulin responses. Male Sprague–Dawley rats were fed either a HFD or low-fat diet (LFD) for 4 weeks. Small resistance artery myograph changes in tension, muscle microvascular recruitment and metabolic response to insulin were determined. Compared with rats fed a LFD, HFD feeding abolished the vasodilatory actions of globular adiponectin (gAd) and insulin on pre-constricted distal saphenous arteries. Pretreatment with gAd improved insulin responses in arterioles isolated from HFD rats, which was blocked by AMP-activated protein kinase (AMPK) inhibition. Similarly, HFD abolished microvascular responses to either gAd or insulin and decreased insulin-stimulated glucose disposal by ∼60%. However, supplementing gAd fully rescued insulin’s microvascular action and significantly improved the metabolic responses to insulin in HFD male rats and these actions were abolished by inhibition of either AMPK or nitric oxide production. We conclude that HFD induces vascular adiponectin and insulin resistance but gAd administration can restore vascular insulin responses and improve insulin’s metabolic action via an AMPK- and nitric oxide-dependent mechanism in male rats. Key points Adiponectin is an adipokine with anti-inflammatory and anti-diabetic properties. Hypoadiponectinaemia is closely associated with endothelial dysfunction and insulin resistance in obesity and diabetes. Insulin resistance is present in muscle microvasculature and this may contribute to decreased insulin delivery to, and action in, muscle. In this study we examined whether adiponectin ameliorates metabolic insulin resistance by affecting muscle

  20. Production of a highly active, soluble form of the cytochrome P450 reductase (CPR A) from Candida tropicalis

    Science.gov (United States)

    Donnelly, Mark

    2006-08-01

    The present invention provides soluble cytochrome p450 reductase (CPR) proteins from Candida sp. having an altered N-terminal region which results in reduced hydrophobicity of the N-terminal region. Also provided are host cells comprising the subject soluble CPR proteins. In addition, the present invention provides nucleotide and corresponding amino acid sequences for soluble CPR proteins and vectors comprising the nucleotide sequences. Methods for producing a soluble CPR, for increasing production of a dicarboxylic acid, and for detecting a cytochrome P450 are also provided.

  1. ESPRIT: A Method for Defining Soluble Expression Constructs in Poorly Understood Gene Sequences.

    Science.gov (United States)

    Mas, Philippe J; Hart, Darren J

    2017-01-01

    Production of soluble, purifiable domains or multi-domain fragments of proteins is a prerequisite for structural biology and other applications. When target sequences are poorly annotated, or when there are few similar sequences available for alignments, identification of domains can be problematic. A method called expression of soluble proteins by random incremental truncation (ESPRIT) addresses this problem by high-throughput automated screening of tens of thousands of enzymatically truncated gene fragments. Rare soluble constructs are identified by experimental screening, and the boundaries revealed by DNA sequencing.

  2. Abalone water-soluble matrix for self-healing biomineralization of tooth defects

    Energy Technology Data Exchange (ETDEWEB)

    Wen, Zhenliang [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Chen, Jingdi, E-mail: ibptcjd@fzu.edu.cn [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Wang, Hailiang [The Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou 350002 (China); Zhong, Shengnan; Hu, Yimin; Wang, Zhili [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Zhang, Qiqing, E-mail: zhangqiq@126.com [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192 (China)

    2016-10-01

    Enamel cannot heal by itself if damaged. Hydroxyapatite (HAP) is main component of human enamel. Formation of enamel-like materials for healing enamel defects remains a challenge. In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% the abalone water-soluble protein (AWSPro) and 2.04 wt% the abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro. Based on X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), hot field emission scanning electron microscopy (HFESEM) and energy dispersive spectrometer (EDS) analysis, the results showed that the AWSM can efficiently induce remineralization of HAP. The enamel-like HAP was successfully achieved onto etched enamel's surface due to the presence of the AWSM. Moreover, the remineralized effect of eroded enamel was growing with the increase of the AWSM. This study provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell, and we provides a new method for self-healing remineralization of enamel defects by AWSM and develops a novel dental material for potential clinical dentistry application. - Graphical abstract: In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% abalone water-soluble protein (AWSPro) and 2.04 wt% abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro by self-organized. Display Omitted - Highlights: • Provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell. • The abalone shell water-soluble matrix contains protein and polysaccharide. • The abalone water-soluble matrix can efficiently induce remineralization of HAP by self-organized. • Achieved self-healing biomineralization of tooth defects in

  3. Abalone water-soluble matrix for self-healing biomineralization of tooth defects

    International Nuclear Information System (INIS)

    Wen, Zhenliang; Chen, Jingdi; Wang, Hailiang; Zhong, Shengnan; Hu, Yimin; Wang, Zhili; Zhang, Qiqing

    2016-01-01

    Enamel cannot heal by itself if damaged. Hydroxyapatite (HAP) is main component of human enamel. Formation of enamel-like materials for healing enamel defects remains a challenge. In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% the abalone water-soluble protein (AWSPro) and 2.04 wt% the abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro. Based on X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), hot field emission scanning electron microscopy (HFESEM) and energy dispersive spectrometer (EDS) analysis, the results showed that the AWSM can efficiently induce remineralization of HAP. The enamel-like HAP was successfully achieved onto etched enamel's surface due to the presence of the AWSM. Moreover, the remineralized effect of eroded enamel was growing with the increase of the AWSM. This study provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell, and we provides a new method for self-healing remineralization of enamel defects by AWSM and develops a novel dental material for potential clinical dentistry application. - Graphical abstract: In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% abalone water-soluble protein (AWSPro) and 2.04 wt% abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro by self-organized. Display Omitted - Highlights: • Provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell. • The abalone shell water-soluble matrix contains protein and polysaccharide. • The abalone water-soluble matrix can efficiently induce remineralization of HAP by self-organized. • Achieved self-healing biomineralization of tooth defects in vitro.

  4. Segmental isotope labeling of proteins for NMR structural study using a protein S tag for higher expression and solubility

    International Nuclear Information System (INIS)

    Kobayashi, Hiroshi; Swapna, G. V. T.; Wu, Kuen-Phon; Afinogenova, Yuliya; Conover, Kenith; Mao, Binchen; Montelione, Gaetano T.; Inouye, Masayori

    2012-01-01

    A common obstacle to NMR studies of proteins is sample preparation. In many cases, proteins targeted for NMR studies are poorly expressed and/or expressed in insoluble forms. Here, we describe a novel approach to overcome these problems. In the protein S tag-intein (PSTI) technology, two tandem 92-residue N-terminal domains of protein S (PrS 2 ) from Myxococcus xanthus is fused at the N-terminal end of a protein to enhance its expression and solubility. Using intein technology, the isotope-labeled PrS 2 -tag is replaced with non-isotope labeled PrS 2 -tag, silencing the NMR signals from PrS 2 -tag in isotope-filtered 1 H-detected NMR experiments. This method was applied to the E. coli ribosome binding factor A (RbfA), which aggregates and precipitates in the absence of a solubilization tag unless the C-terminal 25-residue segment is deleted (RbfAΔ25). Using the PrS 2 -tag, full-length well-behaved RbfA samples could be successfully prepared for NMR studies. PrS 2 (non-labeled)-tagged RbfA (isotope-labeled) was produced with the use of the intein approach. The well-resolved TROSY-HSQC spectrum of full-length PrS 2 -tagged RbfA superimposes with the TROSY-HSQC spectrum of RbfAΔ25, indicating that PrS 2 -tag does not affect the structure of the protein to which it is fused. Using a smaller PrS-tag, consisting of a single N-terminal domain of protein S, triple resonance experiments were performed, and most of the backbone 1 H, 15 N and 13 C resonance assignments for full-length E. coli RbfA were determined. Analysis of these chemical shift data with the Chemical Shift Index and heteronuclear 1 H– 15 N NOE measurements reveal the dynamic nature of the C-terminal segment of the full-length RbfA protein, which could not be inferred using the truncated RbfAΔ25 construct. CS-Rosetta calculations also demonstrate that the core structure of full-length RbfA is similar to that of the RbfAΔ25 construct.

  5. Efficient soluble expression of disulfide bonded proteins in the cytoplasm of Escherichia coli in fed-batch fermentations on chemically defined minimal media.

    Science.gov (United States)

    Gąciarz, Anna; Khatri, Narendar Kumar; Velez-Suberbie, M Lourdes; Saaranen, Mirva J; Uchida, Yuko; Keshavarz-Moore, Eli; Ruddock, Lloyd W

    2017-06-15

    The production of recombinant proteins containing disulfide bonds in Escherichia coli is challenging. In most cases the protein of interest needs to be either targeted to the oxidizing periplasm or expressed in the cytoplasm in the form of inclusion bodies, then solubilized and re-folded in vitro. Both of these approaches have limitations. Previously we showed that soluble expression of disulfide bonded proteins in the cytoplasm of E. coli is possible at shake flask scale with a system, known as CyDisCo, which is based on co-expression of a protein of interest along with a sulfhydryl oxidase and a disulfide bond isomerase. With CyDisCo it is possible to produce disulfide bonded proteins in the presence of intact reducing pathways in the cytoplasm. Here we scaled up production of four disulfide bonded proteins to stirred tank bioreactors and achieved high cell densities and protein yields in glucose fed-batch fermentations, using an E. coli strain (BW25113) with the cytoplasmic reducing pathways intact. Even without process optimization production of purified human single chain IgA 1 antibody fragment reached 139 mg/L and hen avidin 71 mg/L, while purified yields of human growth hormone 1 and interleukin 6 were around 1 g/L. Preliminary results show that human growth hormone 1 was also efficiently produced in fermentations of W3110 strain and when glucose was replaced with glycerol as the carbon source. Our results show for the first time that efficient production of high yields of soluble disulfide bonded proteins in the cytoplasm of E. coli with the reducing pathways intact is feasible to scale-up to bioreactor cultivations on chemically defined minimal media.

  6. A NEW DISTANT MILKY WAY GLOBULAR CLUSTER IN THE PAN-STARRS1 3π SURVEY

    International Nuclear Information System (INIS)

    Laevens, Benjamin P. M.; Martin, Nicolas F.; Sesar, Branimir; Rix, Hans-Walter; Schlafly, Edward F.; Bernard, Edouard J.; Ferguson, Annette M. N.; Slater, Colin T.; Bell, Eric F.; Burgett, William S.; Chambers, Kenneth C.; Denneau, Larry; Kaiser, Nicholas; Kudritzki, Rolf-Peter; Magnier, Eugene A.; Morgan, Jeffrey S.; Sweeney, William E.; Draper, Peter W.; Metcalfe, Nigel; Price, Paul A.

    2014-01-01

    We present a new satellite in the outer halo of the Galaxy, the first Milky Way satellite found in the stacked photometric catalog of the Panoramic Survey Telescope and Rapid Response System 1 (Pan-STARRS1) Survey. From follow-up photometry obtained with WFI on the MPG/ESO 2.2 m telescope, we argue that the object, located at a heliocentric distance of 145 ± 17 kpc, is the most distant Milky Way globular cluster yet known. With a total magnitude of M V = –4.3 ± 0.2 and a half-light radius of 20 ± 2 pc, it shares the properties of extended globular clusters found in the outer halo of our Galaxy and the Andromeda galaxy. The discovery of this distant cluster shows that the full spatial extent of the Milky Way globular cluster system has not yet been fully explored

  7. NO EVIDENCE FOR INTERMEDIATE-MASS BLACK HOLES IN GLOBULAR CLUSTERS: STRONG CONSTRAINTS FROM THE JVLA

    International Nuclear Information System (INIS)

    Strader, Jay; Chomiuk, Laura; Maccarone, Thomas J.; Miller-Jones, James C. A.; Seth, Anil C.; Heinke, Craig O.; Sivakoff, Gregory R.

    2012-01-01

    With a goal of searching for accreting intermediate-mass black holes (IMBHs), we report the results of ultra-deep Jansky Very Large Array radio continuum observations of the cores of three Galactic globular clusters: M15, M19, and M22. We reach rms noise levels of 1.5-2.1 μJy beam –1 at an average frequency of 6 GHz. No sources are observed at the center of any of the clusters. For a conservative set of assumptions about the properties of the accretion, we set 3σ upper limits on IMBHs from 360 to 980 M ☉ . These limits are among the most stringent obtained for any globular cluster. They add to a growing body of work that suggests either (1) IMBHs ∼> 1000 M ☉ are rare in globular clusters or (2) when present, IMBHs accrete in an extraordinarily inefficient manner.

  8. No Evidence for Intermediate-mass Black Holes in Globular Clusters: Strong Constraints from the JVLA

    Science.gov (United States)

    Strader, Jay; Chomiuk, Laura; Maccarone, Thomas J.; Miller-Jones, James C. A.; Seth, Anil C.; Heinke, Craig O.; Sivakoff, Gregory R.

    2012-05-01

    With a goal of searching for accreting intermediate-mass black holes (IMBHs), we report the results of ultra-deep Jansky Very Large Array radio continuum observations of the cores of three Galactic globular clusters: M15, M19, and M22. We reach rms noise levels of 1.5-2.1 μJy beam-1 at an average frequency of 6 GHz. No sources are observed at the center of any of the clusters. For a conservative set of assumptions about the properties of the accretion, we set 3σ upper limits on IMBHs from 360 to 980 M ⊙. These limits are among the most stringent obtained for any globular cluster. They add to a growing body of work that suggests either (1) IMBHs >~ 1000 M ⊙ are rare in globular clusters or (2) when present, IMBHs accrete in an extraordinarily inefficient manner.

  9. pH dependence of the kinetics of interfacial tension changes during protein adsorption from sessile droplets on FEP-Teflon

    NARCIS (Netherlands)

    VanderVegt, W; Norde, W; VanderMei, HC; Busscher, HJ

    Interfacial tension changes during protein adsorption at both the solid-liquid and the liquid-vapor interface were measured simultaneously by ADSA-P from sessile droplets of protein solutions on fluoroethylenepropylene-Teflon. Four globular proteins of similar size, viz. lysozyme, ribonuclease,

  10. Expression by Streptomyces lividans of the Rat α Integrin CD11b A-Domain as a Secreted and Soluble Recombinant Protein

    Directory of Open Access Journals (Sweden)

    Dorra Zouari Ayadi

    2007-01-01

    Full Text Available We already reported the use of a long synthetic signal peptide (LSSP to secrete the Streptomyces sp. TO1 amylase by Streptomyces lividans strain. We herein report the expression and secretion of the rat CD11b A-domain using the same LSSP and S. lividans as host strain. We have used the Escherichia coli/Streptomyces shuttle vector pIJ699 for the cloning of the A-domain DNA sequence downstream of LSSP and under the control of the constitutive ermE-up promoter of Streptomyces erythraeus. Using this construct and S. lividans as a host strain, we achieved the expression of 8 mg/L of soluble secreted recombinant form of the A-domain of the rat leukocyte β2 integrin CD11/CD18 alpha M subunit (CD11b. This secreted recombinant CD11b A-domain reacted with a function blocking antibody showing that this protein is properly folded and probably functional. These data support the capability of Streptomyces to produce heterologous recombinant proteins as soluble secreted form using the “LSSP” synthetic signal peptide.

  11. Analysis of long-range correlation in sequences data of proteins

    OpenAIRE

    ADRIANA ISVORAN; LAURA UNIPAN; DANA CRACIUN; VASILE MORARIU

    2007-01-01

    The results presented here suggest the existence of correlations in the sequence data of proteins. 32 proteins, both globular and fibrous, both monomeric and polymeric, were analyzed. The primary structures of these proteins were treated as time series. Three spatial series of data for each sequence of a protein were generated from numerical correspondences between each amino acid and a physical property associated with it, i.e., its electric charge, its polar character and its dipole moment....

  12. An AO-assisted Variability Study of Four Globular Clusters

    Science.gov (United States)

    Salinas, R.; Contreras Ramos, R.; Strader, J.; Hakala, P.; Catelan, M.; Peacock, M. B.; Simunovic, M.

    2016-09-01

    The image-subtraction technique applied to study variable stars in globular clusters represented a leap in the number of new detections, with the drawback that many of these new light curves could not be transformed to magnitudes due to severe crowding. In this paper, we present observations of four Galactic globular clusters, M 2 (NGC 7089), M 10 (NGC 6254), M 80 (NGC 6093), and NGC 1261, taken with the ground-layer adaptive optics module at the SOAR Telescope, SAM. We show that the higher image quality provided by SAM allows for the calibration of the light curves of the great majority of the variables near the cores of these clusters as well as the detection of new variables, even in clusters where image-subtraction searches were already conducted. We report the discovery of 15 new variables in M 2 (12 RR Lyrae stars and 3 SX Phe stars), 12 new variables in M 10 (11 SX Phe and 1 long-period variable), and 1 new W UMa-type variable in NGC 1261. No new detections are found in M 80, but previous uncertain detections are confirmed and the corresponding light curves are calibrated into magnitudes. Additionally, based on the number of detected variables and new Hubble Space Telescope/UVIS photometry, we revisit a previous suggestion that M 80 may be the globular cluster with the richest population of blue stragglers in our Galaxy. Based on observations obtained at the Southern Astrophysical Research (SOAR) telescope, which is a joint project of the Ministério da Ciência, Tecnologia, e Inovação (MCTI) da República Federativa do Brasil, the U.S. National Optical Astronomy Observatory (NOAO), the University of North Carolina at Chapel Hill (UNC), and Michigan State University (MSU).

  13. alpha isoforms of soluble and membrane-linked folate-binding protein in human blood

    DEFF Research Database (Denmark)

    Hoier-Madsen, M.; Holm, J.; Hansen, S.I.

    2008-01-01

    supported the hypothesis that serum FBP (29 kDa) mainly originates from neutrophils. The presence of FBP/FR alpha isoforms were established for the first time in human blood using antibodies specifically directed against human milk FBP alpha. The alpha isoforms identified on erythrocyte membranes......, and in granulocytes and serum, only constituted an almost undetectable fraction of the functional FBP The FBP alpha in neutrophil granulocytes was identified as a cytoplasmic component by indirect immunofluorescence. Gel filtration of serum revealed a peak of FBP alpha (>120 kDa), which could represent receptor...... fragments from decomposed erythrocytes and granulocytes. The soluble FBPs may exert bacteriostatic effects and protect folates in plasma from biological degradation, whereas FRs on the surface of blood cells could be involved in intracellular folate uptake or serve as signal proteins. The latter receptors...

  14. Course 12: Proteins: Structural, Thermodynamic and Kinetic Aspects

    Science.gov (United States)

    Finkelstein, A. V.

    1 Introduction 2 Overview of protein architectures and discussion of physical background of their natural selection 2.1 Protein structures 2.2 Physical selection of protein structures 3 Thermodynamic aspects of protein folding 3.1 Reversible denaturation of protein structures 3.2 What do denatured proteins look like? 3.3 Why denaturation of a globular protein is the first-order phase transition 3.4 "Gap" in energy spectrum: The main characteristic that distinguishes protein chains from random polymers 4 Kinetic aspects of protein folding 4.1 Protein folding in vivo 4.2 Protein folding in vitro (in the test-tube) 4.3 Theory of protein folding rates and solution of the Levinthal paradox

  15. A Visual Language for Protein Design

    KAUST Repository

    Cox, Robert Sidney

    2017-02-08

    As protein engineering becomes more sophisticated, practitioners increasingly need to share diagrams for communicating protein designs. To this end, we present a draft visual language, Protein Language, that describes the high-level architecture of an engineered protein with easy-to draw glyphs, intended to be compatible with other biological diagram languages such as SBOL Visual and SBGN. Protein Language consists of glyphs for representing important features (e.g., globular domains, recognition and localization sequences, sites of covalent modification, cleavage and catalysis), rules for composing these glyphs to represent complex architectures, and rules constraining the scaling and styling of diagrams. To support Protein Language we have implemented an extensible web-based software diagram tool, Protein Designer, that uses Protein Language in a

  16. A Visual Language for Protein Design

    KAUST Repository

    Cox, Robert Sidney; McLaughlin, James Alastair; Grunberg, Raik; Beal, Jacob; Wipat, Anil; Sauro, Herbert M.

    2017-01-01

    As protein engineering becomes more sophisticated, practitioners increasingly need to share diagrams for communicating protein designs. To this end, we present a draft visual language, Protein Language, that describes the high-level architecture of an engineered protein with easy-to draw glyphs, intended to be compatible with other biological diagram languages such as SBOL Visual and SBGN. Protein Language consists of glyphs for representing important features (e.g., globular domains, recognition and localization sequences, sites of covalent modification, cleavage and catalysis), rules for composing these glyphs to represent complex architectures, and rules constraining the scaling and styling of diagrams. To support Protein Language we have implemented an extensible web-based software diagram tool, Protein Designer, that uses Protein Language in a

  17. Revealing the mechanisms of protein disorder and N-glycosylation in CD44-hyaluronan binding using molecular simulation

    Directory of Open Access Journals (Sweden)

    Olgun eGuvench

    2015-06-01

    Full Text Available The extracellular N-terminal hyaluronan binding domain (HABD of CD44 is a small globular domain that confers hyaluronan (HA binding functionality to this large transmembrane glycoprotein. When recombinantly expressed by itself, HABD exists as a globular water-soluble protein that retains the capacity to bind HA. This has enabled atomic-resolution structural biology experiments that have revealed the structure of HABD and its binding mode with oligomeric HA. Such experiments have also pointed to an order-to-disorder transition in HABD that is associated with HA binding. However, it had remained unclear how this structural transition was involved in binding since it occurs in a region of HABD distant from the HA-binding site. Furthermore, HABD is known to be N-glycosylated, and such glycosylation can diminish HA binding when the associated N-glycans are capped with sialic acid residues. The intrinsic flexibility of disordered proteins and of N-glycans makes it difficult to apply experimental structural biology approaches to probe the molecular mechanisms of how the order-to-disorder transition and N-glycosylation can modulate HA binding by HABD. We review recent results from molecular dynamics simulations that provide atomic-resolution mechanistic understanding of such modulation to help bridge gaps between existing experimental binding and structural biology data. Findings from these simulations include: Tyr42 may function as a molecular switch that converts the HA binding site from a low affinity to a high affinity state; in the partially-disordered form of HABD, basic amino acids in the C-terminal region can gain sufficient mobility to form direct contacts with bound HA to further stabilize binding; and terminal sialic acids on covalently-attached N-glycans can form charge-paired hydrogen bonding interactions with basic amino acids that could otherwise bind to HA, thereby blocking HA binding to glycosylated CD44 HABD.

  18. BVRI CCD photometry of the globular cluster NGC 2808

    International Nuclear Information System (INIS)

    Alcaino, G.; Liller, W.; Alvarado, F.; Wenderoth, E.

    1990-01-01

    As a part of a continuing program, CCD color-magnitude diagrams are presented for the bright globular cluster NGC 2808 in the four colors comprising BVRI. From a comparison of four different CMDs with theoretical isochrones, an age of 16 + or - 2 Gyr is obtained, assuming a value for Fe/H near -1.3. 28 refs

  19. Expression and purification of short hydrophobic elastin-like polypeptides with maltose-binding protein as a solubility tag.

    Science.gov (United States)

    Bataille, Laure; Dieryck, Wilfrid; Hocquellet, Agnès; Cabanne, Charlotte; Bathany, Katell; Lecommandoux, Sébastien; Garbay, Bertrand; Garanger, Elisabeth

    2015-06-01

    Elastin-like polypeptides (ELPs) are biodegradable polymers with interesting physico-chemical properties for biomedical and biotechnological applications. The recombinant expression of hydrophobic elastin-like polypeptides is often difficult because they possess low transition temperatures, and therefore form aggregates at sub-ambient temperatures. To circumvent this difficulty, we expressed in Escherichia coli three hydrophobic ELPs (VPGIG)n with variable lengths (n=20, 40, and 60) in fusion with the maltose-binding protein (MBP). Fusion proteins were soluble and yields of purified MBP-ELP ranged between 66 and 127mg/L culture. After digestion of the fusion proteins by enterokinase, the ELP moiety was purified by using inverse transition cycling. The purified fraction containing ELP40 was slightly contaminated by traces of undigested fusion protein. Purification of ELP60 was impaired because of co-purification of the MBP tag during inverse transition cycling. ELP20 was successfully purified to homogeneity, as assessed by gel electrophoresis and mass spectrometry analyses. The transition temperature of ELP20 was measured at 15.4°C in low salt buffer. In conclusion, this method can be used to produce hydrophobic ELP of low molecular mass. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Light-fuelled transport of large dendrimers and proteins.

    Science.gov (United States)

    Koskela, Jenni E; Liljeström, Ville; Lim, Jongdoo; Simanek, Eric E; Ras, Robin H A; Priimagi, Arri; Kostiainen, Mauri A

    2014-05-14

    This work presents a facile water-based supramolecular approach for light-induced surface patterning. The method is based upon azobenzene-functionalized high-molecular weight triazine dendrimers up to generation 9, demonstrating that even very large globular supramolecular complexes can be made to move in response to light. We also demonstrate light-fuelled macroscopic movements in native biomolecules, showing that complexes of apoferritin protein and azobenzene can effectively form light-induced surface patterns. Fundamentally, the results establish that thin films comprising both flexible and rigid globular particles of large diameter can be moved with light, whereas the presented material concepts offer new possibilities for the yet marginally explored biological applications of azobenzene surface patterning.

  1. From Globular Clusters to Tidal Dwarfs: Structure Formation in Tidal Tails

    Science.gov (United States)

    Knierman, K.; Hunsberger, S.; Gallagher, S.; Charlton, J.; Whitmore, B.; Hibbard, J.; Kundu, A.; Zaritsky, D.

    1999-12-01

    Galaxy interactions trigger star formation in tidal debris. How does this star formation depend on the local and global physical conditions? Using WFPC2/HST images, we investigate the range of structure within tidal tails of four classic ``Toomre Sequence'' mergers: NGC 4038/9 (``Antennae''), NGC 7252 (``Atoms for Peace''), NGC 3921, and NGC 3256. These tails contain a variety of stellar associations with sizes from globular clusters up to dwarf Irregulars. We explore whether there is a continuum between the two extremes. Our eight fields sample seven tidal tails at a variety of stages in the evolutionary sequence. Some of these tails are rich in HI while others are HI poor. Large tidal dwarfs are embedded in three of the tails. Using V and I WFPC2 images, we measure luminosities and colors of substructures within the tidal tails. The properties of globular cluster candidates in the tails will be contrasted with those of the hundreds of young clusters in the central regions of these mergers. We address whether globular clusters form and survive in the tidal tails and whether tidal dwarfs are composed of only young stars. By comparing the properties of structures in the tails of the four mergers with different ages, we examine systematic evolution of structure along the evolutionary sequence and as a function of HI content. We acknowledge support from NASA through STScI, and from NSF for an REU supplement for Karen Knierman.

  2. Silk Spinning in Silkworms and Spiders.

    Science.gov (United States)

    Andersson, Marlene; Johansson, Jan; Rising, Anna

    2016-08-09

    Spiders and silkworms spin silks that outcompete the toughness of all natural and manmade fibers. Herein, we compare and contrast the spinning of silk in silkworms and spiders, with the aim of identifying features that are important for fiber formation. Although spiders and silkworms are very distantly related, some features of spinning silk seem to be universal. Both spiders and silkworms produce large silk proteins that are highly repetitive and extremely soluble at high pH, likely due to the globular terminal domains that flank an intermediate repetitive region. The silk proteins are produced and stored at a very high concentration in glands, and then transported along a narrowing tube in which they change conformation in response primarily to a pH gradient generated by carbonic anhydrase and proton pumps, as well as to ions and shear forces. The silk proteins thereby convert from random coil and alpha helical soluble conformations to beta sheet fibers. We suggest that factors that need to be optimized for successful production of artificial silk proteins capable of forming tough fibers include protein solubility, pH sensitivity, and preservation of natively folded proteins throughout the purification and initial spinning processes.

  3. Coarse-grain modelling of protein-protein interactions

    NARCIS (Netherlands)

    Baaden, Marc; Marrink, Siewert J.

    2013-01-01

    Here, we review recent advances towards the modelling of protein-protein interactions (PPI) at the coarse-grained (CG) level, a technique that is now widely used to understand protein affinity, aggregation and self-assembly behaviour. PPI models of soluble proteins and membrane proteins are

  4. Structural insights into the globular tails of the human type v myosins Myo5a, Myo5b, And Myo5c.

    Directory of Open Access Journals (Sweden)

    Hana Velvarska

    Full Text Available Vertebrate type V myosins (MyoV Myo5a, Myo5b, and Myo5c mediate transport of several different cargoes. All MyoV paralogs bind to cargo complexes mainly by their C-terminal globular domains. In absence of cargo, the globular domain of Myo5a inhibits its motor domain. Here, we report low-resolution SAXS models for the globular domains from human Myo5a, Myo5b, and Myo5c, which suggest very similar overall shapes of all three paralogs. We determined the crystal structures of globular domains from Myo5a and Myo5b, and provide a homology model for human Myo5c. When we docked the Myo5a crystal structure into a previously published electron microscopy density of the autoinhibited full-length Myo5a, only one domain orientation resulted in a good fit. This structural arrangement suggests the participation of additional region of the globular domain in autoinhibition. Quantification of the interaction of the Myo5a globular domain with its motor complex revealed a tight binding with dissociation half-life in the order of minutes, suggesting a rather slow transition between the active and inactive states.

  5. Soluble polymorphic bank vole prion proteins induced by co-expression of quiescin sulfhydryl oxidase in E. coli and their aggregation behaviors.

    Science.gov (United States)

    Abskharon, Romany; Dang, Johnny; Elfarash, Ameer; Wang, Zerui; Shen, Pingping; Zou, Lewis S; Hassan, Sedky; Wang, Fei; Fujioka, Hisashi; Steyaert, Jan; Mulaj, Mentor; Surewicz, Witold K; Castilla, Joaquín; Wohlkonig, Alexandre; Zou, Wen-Quan

    2017-10-04

    The infectious prion protein (PrP Sc or prion) is derived from its cellular form (PrP C ) through a conformational transition in animal and human prion diseases. Studies have shown that the interspecies conversion of PrP C to PrP Sc is largely swayed by species barriers, which is mainly deciphered by the sequence and conformation of the proteins among species. However, the bank vole PrP C (BVPrP) is highly susceptible to PrP Sc from different species. Transgenic mice expressing BVPrP with the polymorphic isoleucine (109I) but methionine (109M) at residue 109 spontaneously develop prion disease. To explore the mechanism underlying the unique susceptibility and convertibility, we generated soluble BVPrP by co-expression of BVPrP with Quiescin sulfhydryl oxidase (QSOX) in Escherichia coli. Interestingly, rBVPrP-109M and rBVPrP-109I exhibited distinct seeded aggregation pathways and aggregate morphologies upon seeding of mouse recombinant PrP fibrils, as monitored by thioflavin T fluorescence and electron microscopy. Moreover, they displayed different aggregation behaviors induced by seeding of hamster and mouse prion strains under real-time quaking-induced conversion. Our results suggest that QSOX facilitates the formation of soluble prion protein and provide further evidence that the polymorphism at residue 109 of QSOX-induced BVPrP may be a determinant in mediating its distinct convertibility and susceptibility.

  6. Chemical analysis of eight giant stars of the globular cluster NGC 6366

    Science.gov (United States)

    Puls, Arthur A.; Alves-Brito, Alan; Campos, Fabíola; Dias, Bruno; Barbuy, Beatriz

    2018-05-01

    The metal-rich Galactic globular cluster NGC 6366 is the fifth closest to the Sun. Despite its interest, it has received scarce attention, and little is known about its internal structure. Its kinematics suggests a link to the halo, but its metallicity indicates otherwise. We present a detailed chemical analysis of eight giant stars of NGC 6366, using high-resolution and high-quality spectra (R > 40 000, S/N > 60) obtained at the VLT (8.2 m) and CFHT (3.6 m) telescopes. We attempted to characterize its chemistry and to search for evidence of multiple stellar populations. The atmospheric parameters were derived using the method of excitation and ionization equilibrium of Fe I and Fe II lines and from those atmospheric parameters we calculated the abundances for other elements and found that none of the elements measured presents star-to-star variation greater than the uncertainties. We compared the derived abundances with those of other globular clusters and field stars available in the literature. We determined a mean [Fe/H] = -0.60 ± 0.03 for NGC 6366 and found some similarity of this object with M 71, another inner halo globular cluster. The Na-O anticorrelation extension is short and no star-to-star variation in Al is found. The presence of second generation stars is not evident in NGC 6366.

  7. pH-sensitive polymeric nanoparticles to improve oral bioavailability of peptide/protein drugs and poorly water-soluble drugs.

    Science.gov (United States)

    Wang, Xue-Qing; Zhang, Qiang

    2012-10-01

    pH-sensitive polymeric nanoparticles are promising for oral drug delivery, especially for peptide/protein drugs and poorly water-soluble medicines. This review describes current status of pH-sensitive polymeric nanoparticles for oral drug delivery and introduces the mechanisms of drug release from them as well as possible reasons for absorption improvement, with emphasis on our contribution to this field. pH-sensitive polymeric nanoparticles are prepared mainly with polyanions, polycations, their mixtures or cross-linked polymers. The mechanisms of drug release are the result of carriers' dissolution, swelling or both of them at specific pH. The possible reasons for improvement of oral bioavailability include the following: improve drug stability, enhance mucoadhesion, prolong resident time in GI tract, ameliorate intestinal permeability and increase saturation solubility and dissolution rate for poorly water-soluble drugs. As for the advantages of pH-sensitive nanoparticles over conventional nanoparticles, we conclude that (1) most carriers used are enteric-coating materials and their safety has been approved. (2) The rapid dissolution or swelling of carriers at specific pH results in quick drug release and high drug concentration gradient, which is helpful for absorption. (3) At the specific pH carriers dissolve or swell, and the bioadhesion of carriers to mucosa becomes high because nanoparticles turn from solid to gel, which can facilitate drug absorption. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Effects of Acids, Bases, and Heteroatoms on Proximal Radial Distribution Functions for Proteins.

    Science.gov (United States)

    Nguyen, Bao Linh; Pettitt, B Montgomery

    2015-04-14

    The proximal distribution of water around proteins is a convenient method of quantifying solvation. We consider the effect of charged and sulfur-containing amino acid side-chain atoms on the proximal radial distribution function (pRDF) of water molecules around proteins using side-chain analogs. The pRDF represents the relative probability of finding any solvent molecule at a distance from the closest or surface perpendicular protein atom. We consider the near-neighbor distribution. Previously, pRDFs were shown to be universal descriptors of the water molecules around C, N, and O atom types across hundreds of globular proteins. Using averaged pRDFs, a solvent density around any globular protein can be reconstructed with controllable relative error. Solvent reconstruction using the additional information from charged amino acid side-chain atom types from both small models and protein averages reveals the effects of surface charge distribution on solvent density and improves the reconstruction errors relative to simulation. Solvent density reconstructions from the small-molecule models are as effective and less computationally demanding than reconstructions from full macromolecular models in reproducing preferred hydration sites and solvent density fluctuations.

  9. Effect of tissue scaffold topography on protein structure monitored by fluorescence spectroscopy.

    Science.gov (United States)

    Portugal, Carla A M; Truckenmüller, Roman; Stamatialis, Dimitrios; Crespo, João G

    2014-11-10

    The impact of surface topography on the structure of proteins upon adhesion was assessed through non-invasive fluorescence monitoring. This study aimed at obtaining a better understanding about the role of protein structural status on cell-scaffold interactions. The changes induced upon adsorption of two model proteins with different geometries, trypsin (globular conformation) and fibrinogen (rod-shaped conformation) on poly-l-lactic acid (PLLA) scaffolds with different surface topographies, flat, fibrous and surfaces with aligned nanogrooves, were assessed by fluorescence spectroscopy monitoring, using tryptophan as structural probe. Hence, the maximum emission blue shift and the increase of fluorescence anisotropy observed after adsorption of globular and rod-like shaped proteins on surfaces with parallel nanogrooves were ascribed to more intense protein-surface interactions. Furthermore, the decrease of fluorescence anisotropy observed upon adsorption of proteins to scaffolds with fibrous morphology was more significant for rod-shaped proteins. This effect was associated to the ability of these proteins to adjust to curved surfaces. The additional unfolding of proteins induced upon adsorption on scaffolds with a fibrous morphology may be the reason for better cell attachment there, promoting an easier access of cell receptors to initially hidden protein regions (e.g. RGDS sequence), which are known to have a determinant role in cell attaching processes. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. THE IMPACT OF CONTAMINATED RR LYRAE/GLOBULAR CLUSTER PHOTOMETRY ON THE DISTANCE SCALE

    Energy Technology Data Exchange (ETDEWEB)

    Majaess, D.; Turner, D.; Lane, D. [Department of Astronomy and Physics, Saint Mary' s University, Halifax, NS B3H 3C3 (Canada); Gieren, W., E-mail: dmajaess@ap.smu.ca [Departamento de Astronomia, Universidad de Concepcion, Casilla 160-C, CL Concepcion (Chile)

    2012-06-10

    RR Lyrae variables and the stellar constituents of globular clusters are employed to establish the cosmic distance scale and age of the universe. However, photometry for RR Lyrae variables in the globular clusters M3, M15, M54, M92, NGC 2419, and NGC 6441 exhibit a dependence on the clustercentric distance. For example, variables and stars positioned near the crowded high-surface brightness cores of the clusters may suffer from photometric contamination, which invariably affects a suite of inferred parameters (e.g., distance, color excess, absolute magnitude, etc.). The impetus for this study is to mitigate the propagation of systematic uncertainties by increasing awareness of the pernicious impact of contaminated and radial-dependent photometry.

  11. M31 GLOBULAR CLUSTER STRUCTURES AND THE PRESENCE OF X-RAY BINARIES

    International Nuclear Information System (INIS)

    Agar, J. R. R.; Barmby, P.

    2013-01-01

    The Andromeda galaxy, M31, has several times the number of globular clusters found in the Milky Way. It contains a correspondingly larger number of low-mass X-ray binaries (LMXBs) associated with globular clusters, and as such can be used to investigate the cluster properties that lead to X-ray binary formation. The best tracer of the spatial structure of M31 globulars is the high-resolution imaging available from the Hubble Space Telescope (HST), and we have used HST data to derive structural parameters for 29 LMXB-hosting M31 globular clusters. These measurements are combined with structural parameters from the literature for a total of 41 (of 50 known) LMXB clusters and a comparison sample of 65 non-LMXB clusters. Structural parameters measured in blue bandpasses are found to be slightly different (smaller core radii and higher concentrations) than those measured in red bandpasses; this difference is enhanced in LMXB clusters and could be related to stellar population differences. Clusters with LMXBs show higher collision rates for their mass compared to clusters without LMXBs, and collision rates estimated at the core radius show larger offsets than rates estimated at the half-light radius. These results are consistent with the dynamical formation scenario for LMXBs. A logistic regression analysis finds that, as expected, the probability of a cluster hosting an LMXB increases with increasing collision rate and proximity to the galaxy center. The same analysis finds that probability of a cluster hosting an LMXB decreases with increasing cluster mass at a fixed collision rate, although we caution that this could be due to sample selection effects. Metallicity is found to be a less important predictor of LMXB probability than collision rate, mass, or distance, even though LMXB clusters have a higher metallicity on average. This may be due to the interaction of location and metallicity: a sample of M31 LMXBs with a greater range in galactocentric distance would

  12. LITHIUM-RICH GIANTS IN GLOBULAR CLUSTERS

    Energy Technology Data Exchange (ETDEWEB)

    Kirby, Evan N.; Cohen, Judith G. [California Institute of Technology, 1200 E. California Boulevard, MC 249-17, Pasadena, CA 91125 (United States); Guhathakurta, Puragra [UCO/Lick Observatory and Department of Astronomy and Astrophysics, University of California, 1156 High Street, Santa Cruz, CA 95064 (United States); Zhang, Andrew J. [The Harker School, 500 Saratoga Avenue, San Jose, CA 95129 (United States); Hong, Jerry [Palo Alto High School, 50 Embarcadero Road, Palo Alto, CA, 94301 (United States); Guo, Michelle [Stanford University, 450 Serra Mall, Stanford, CA 94305 (United States); Guo, Rachel [Irvington High School, 41800 Blacow Road, Fremont, CA 94538 (United States); Cunha, Katia [Observatório Nacional, São Cristóvão Rio de Janeiro (Brazil)

    2016-03-10

    Although red giants deplete lithium on their surfaces, some giants are Li-rich. Intermediate-mass asymptotic giant branch (AGB) stars can generate Li through the Cameron–Fowler conveyor, but the existence of Li-rich, low-mass red giant branch (RGB) stars is puzzling. Globular clusters are the best sites to examine this phenomenon because it is straightforward to determine membership in the cluster and to identify the evolutionary state of each star. In 72 hours of Keck/DEIMOS exposures in 25 clusters, we found four Li-rich RGB and two Li-rich AGB stars. There were 1696 RGB and 125 AGB stars with measurements or upper limits consistent with normal abundances of Li. Hence, the frequency of Li-richness in globular clusters is (0.2 ± 0.1)% for the RGB, (1.6 ± 1.1)% for the AGB, and (0.3 ± 0.1)% for all giants. Because the Li-rich RGB stars are on the lower RGB, Li self-generation mechanisms proposed to occur at the luminosity function bump or He core flash cannot explain these four lower RGB stars. We propose the following origin for Li enrichment: (1) All luminous giants experience a brief phase of Li enrichment at the He core flash. (2) All post-RGB stars with binary companions on the lower RGB will engage in mass transfer. This scenario predicts that 0.1% of lower RGB stars will appear Li-rich due to mass transfer from a recently Li-enhanced companion. This frequency is at the lower end of our confidence interval.

  13. Biological variation and reference intervals for circulating osteopontin, osteoprotegerin, total soluble receptor activator of nuclear factor kappa B ligand and high-sensitivity C-reactive protein

    DEFF Research Database (Denmark)

    Sennels, H P; Jacobsen, Søren; Jensen, T

    2007-01-01

    Objective. Monitoring inflammatory diseases and osteoclastogenesis with osteopontin (OPN), osteoprotegerin (OPG), total soluble receptor activator of nuclear factor kappa B ligand (total sRANKL) and high-sensitivity C-reactive protein (hsCRP) has recently attracted increased interest. The purpose...

  14. The deposition of globular polypyrrole and polypyrrole nanotubes on cotton textile

    Czech Academy of Sciences Publication Activity Database

    Bober, Patrycja; Stejskal, Jaroslav; Šeděnková, Ivana; Trchová, Miroslava; Martinková, L.; Marek, J.

    2015-01-01

    Roč. 356, 30 November (2015), s. 737-741 ISSN 0169-4332 R&D Projects: GA TA ČR(CZ) TE01020022 Institutional support: RVO:61389013 Keywords : conducting textile * cotton * globular polypyrrole Subject RIV: CG - Electrochemistry Impact factor: 3.150, year: 2015

  15. Deep CCD photometry in globular clusters III. M15

    International Nuclear Information System (INIS)

    Fahlman, G.G.; Richer, H.B.; Vandenberg, D.A.

    1985-01-01

    CCD photometry in U, B, and V is presented for a 5' x 3' field in the globular cluster M15. The location of the main sequence in the color-magnitude diagram is found here to be significantly bluer than previous studies have indicated. The luminosity function of the cluster is studied down to V = 22.8 (Mroughly-equal7.5) and shown to be consistent with a power-law mass function, n(M) = QM/sup -alpha/ with α = 2.5 +- 1.0, to the limit of our data. The field star population brighter than V = 21.5, is examined in some detail. There appears to be about 50% more stars belonging to the disk in the field as compared with the Bahcall-Soneira standard galaxy model. The reddening to the cluster is found to be E(B-V) = 0.11 +- 0.04 from nine bright field stars. A new value for the ultraviolet excess of the cluster main-sequence stars is obtained, delta(0.6) = 0.25 +- 0.02, and confirms the well-known fact that M15 is among the metal poorest of the globular clusters

  16. An Archival Search For Young Globular Clusters in Galaxies

    Science.gov (United States)

    Whitmore, Brad

    1995-07-01

    One of the most intriguing results from HST has been the discovery of ultraluminous star clusters in interacting and merging galaxies. These clusters have the luminosities, colors, and sizes that would be expected of young globular clusters produced by the interaction. We propose to use the data in the HST Archive to determine how prevalent this phenomena is, and to determine whether similar clusters are produced in other environments. Three samples will be extracted and studied in a systematic and consistent manner: 1} interacting and merging galaxies, 2} starburst galaxies, 3} a control sample of ``normal'' galaxies. A preliminary search of the archives shows that there are at least 20 galaxies in each of these samples, and the number will grow by about 50 observations become available. The data will be used to determine the luminosity function, color histogram , spatial distribution, and structural properties of the clusters using the same techniques employed in our study of NGC 7252 {``Atoms -for-Peace'' galaxy} and NGC 4038/4039 {``The Antennae''}. Our ultimate goals are: 1} to understand how globular clusters form, and 2} to use the clusters as evolutionary tracers to unravel the histories of interacting galaxies.

  17. Deep radio synthesis images of globular clusters

    International Nuclear Information System (INIS)

    Kulkarni, S.R.; Goss, W.M.; Wolszczan, A.; Middleditch, J.

    1990-01-01

    Results are reported from a program of high-resolution and high-sensitivity imaging of globular clusters at 20 cm. The findings indicate that there is not a large number of pulsars in compact binaries which have escaped detection in single-dish pulse searches. Such binaries have been postulated to result from tidal captures of single main-sequence stars. It is suggested that most tidal captures involving neutron stars ultimately result in the formation of a spun-up single pulsar and the complete disruption of the main-sequence star. 27 refs

  18. Soluble N-Ethylmaleimide-Sensitive Factor Attachment Protein Receptor-Derived Peptides for Regulation of Mast Cell Degranulation.

    Science.gov (United States)

    Yang, Yoosoo; Kong, Byoungjae; Jung, Younghoon; Park, Joon-Bum; Oh, Jung-Mi; Hwang, Jaesung; Cho, Jae Youl; Kweon, Dae-Hyuk

    2018-01-01

    Vesicle-associated V-soluble N -ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins and target membrane-associated T-SNAREs (syntaxin 4 and SNAP-23) assemble into a core trans -SNARE complex that mediates membrane fusion during mast cell degranulation. This complex plays pivotal roles at various stages of exocytosis from the initial priming step to fusion pore opening and expansion, finally resulting in the release of the vesicle contents. In this study, peptides with the sequences of various SNARE motifs were investigated for their potential inhibitory effects against SNARE complex formation and mast cell degranulation. The peptides with the sequences of the N-terminal regions of vesicle-associated membrane protein 2 (VAMP2) and VAMP8 were found to reduce mast cell degranulation by inhibiting SNARE complex formation. The fusion of protein transduction domains to the N-terminal of each peptide enabled the internalization of the fusion peptides into the cells equally as efficiently as cell permeabilization by streptolysin-O without any loss of their inhibitory activities. Distinct subsets of mast cell granules could be selectively regulated by the N-terminal-mimicking peptides derived from VAMP2 and VAMP8, and they effectively decreased the symptoms of atopic dermatitis in mouse models. These results suggest that the cell membrane fusion machinery may represent a therapeutic target for atopic dermatitis.

  19. Variable Stars in Large Magellanic Cloud Globular Clusters. II. NGC 1786

    Science.gov (United States)

    Kuehn, Charles A.; Smith, Horace A.; Catelan, Márcio; Pritzl, Barton J.; De Lee, Nathan; Borissova, Jura

    2012-12-01

    This is the second in a series of papers studying the variable stars in Large Magellanic Cloud globular clusters. The primary goal of this series is to study how RR Lyrae stars in Oosterhoff-intermediate systems compare to their counterparts in Oosterhoff I/II systems. In this paper, we present the results of our new time-series B-V photometric study of the globular cluster NGC 1786. A total of 65 variable stars were identified in our field of view. These variables include 53 RR Lyraes (27 RRab, 18 RRc, and 8 RRd), 3 classical Cepheids, 1 Type II Cepheid, 1 Anomalous Cepheid, 2 eclipsing binaries, 3 Delta Scuti/SX Phoenicis variables, and 2 variables of undetermined type. Photometric parameters for these variables are presented. We present physical properties for some of the RR Lyrae stars, derived from Fourier analysis of their light curves. We discuss several different indicators of Oosterhoff type which indicate that the Oosterhoff classification of NGC 1786 is not as clear cut as what is seen in most globular clusters. Based on observations taken with the SMARTS 1.3 m telescope operated by the SMARTS Consortium and observations taken at the Southern Astrophysical Research (SOAR) telescope, which is a joint project of the Ministério da Ciência, Tecnologia, e Inovação (MCTI) da República Federativa do Brasil, the U.S. National Optical Astronomy Observatory (NOAO), the University of North Carolina at Chapel Hill (UNC), and Michigan State University (MSU).

  20. The "Globularization Hypothesis" of the Language-ready Brain as a Developmental Frame for Prosodic Bootstrapping Theories of Language Acquisition.

    Science.gov (United States)

    Irurtzun, Aritz

    2015-01-01

    In recent research (Boeckx and Benítez-Burraco, 2014a,b) have advanced the hypothesis that our species-specific language-ready brain should be understood as the outcome of developmental changes that occurred in our species after the split from Neanderthals-Denisovans, which resulted in a more globular braincase configuration in comparison to our closest relatives, who had elongated endocasts. According to these authors, the development of a globular brain is an essential ingredient for the language faculty and in particular, it is the centrality occupied by the thalamus in a globular brain that allows its modulatory or regulatory role, essential for syntactico-semantic computations. Their hypothesis is that the syntactico-semantic capacities arise in humans as a consequence of a process of globularization, which significantly takes place postnatally (cf. Neubauer et al., 2010). In this paper, I show that Boeckx and Benítez-Burraco's hypothesis makes an interesting developmental prediction regarding the path of language acquisition: it teases apart the onset of phonological acquisition and the onset of syntactic acquisition (the latter starting significantly later, after globularization). I argue that this hypothesis provides a developmental rationale for the prosodic bootstrapping hypothesis of language acquisition (cf. i.a. Gleitman and Wanner, 1982; Mehler et al., 1988, et seq.; Gervain and Werker, 2013), which claim that prosodic cues are employed for syntactic parsing. The literature converges in the observation that a large amount of such prosodic cues (in particular, rhythmic cues) are already acquired before the completion of the globularization phase, which paves the way for the premises of the prosodic bootstrapping hypothesis, allowing babies to have a rich knowledge of the prosody of their target language before they can start parsing the primary linguistic data syntactically.

  1. The globularization hypothesis of the language-ready brain as a developmental frame for prosodic bootstrapping theories of language acquisition

    Directory of Open Access Journals (Sweden)

    Aritz eIrurtzun

    2015-12-01

    Full Text Available In recent research Boeckx & Benítez-Burraco (2014a,b have advanced the hypothesis that our species-specific language-ready brain should be understood as the outcome of developmental changes that occurred in our species after the split from Neanderthals-Denisovans, which resulted in a more globular braincase configuration in comparison to our closest relatives, who had elongated endocasts. According to these authors, the development of a globular brain is an essential ingredient for the language faculty and in particular, it is the centrality occupied by the thalamus in a globular brain that allows its modulatory or regulatory role, essential for syntactico-semantic computations. Their hypothesis is that the syntactico-semantic capacities arise in humans as a consequence of a process of globularization, which significantly takes place postnatally (cf. Neubauer et al. (2010. In this paper, I show that Boeckx & Benítez-Burraco’s hypothesis makes an interesting developmental prediction regarding the path of language acquisition: it teases apart the onset of phonological acquisition and the onset of syntactic acquisition (the latter starting significantly later, after globularization. I argue that this hypothesis provides a developmental rationale for the prosodic bootstrapping hypothesis of language acquisition (cf. i.a. Gleitman & Wanner (1982; Mehler et al. (1988, et seq.; Gervain & Werker (2013, which claim that prosodic cues are employed for syntactic parsing. The literature converges in the observation that a large amount of such prosodic cues (in particular, rhythmic cues are already acquired before the completion of the globularization phase, which paves the way for the premises of prosodic bootstrapping hypothesis, allowing babies to have a rich knowledge of the prosody of their target language before they can start parsing the primary linguistic data syntactically.

  2. Pretreatment with soluble ST2 reduces warm hepatic ischemia/reperfusion injury

    International Nuclear Information System (INIS)

    Yin Hui; Huang Baojun; Yang Heng; Huang Yafei; Xiong Ping; Zheng Fang; Chen Xiaoping; Chen Yifa; Gong Feili

    2006-01-01

    The interleukin-1 receptor-like protein ST2 exists in both membrane-bound (ST2L) and soluble form (sST2). ST2L has been found to play an important regulatory role in Th2-type immune response, but the function of soluble form of ST2 remains to be elucidated. In this study, we report the protective effect of soluble ST2 on warm hepatic ischemia/reperfusion injury. We constructed a eukaryotic expression plasmid, psST2-Fc, which expresses functional murine soluble ST2-human IgG1 Fc (sST2-Fc) fusion protein. The liver damage after ischemia/reperfusion was significantly attenuated by the expression of this plasmid in vivo. sST2-Fc remarkably inhibited the activation of Kupffer cells and the production of proinflammatory mediators TNF-α and IL-6. Furthermore, the levels of TLR4 mRNA and the nuclear translocation of NF-κB were also suppressed by pretreatment with sST2-Fc. These results thus identified soluble ST2 as a negative regulator in hepatic I/R injury, possibly via ST2-TLR4 pathway

  3. Development and Fit-for-Purpose Validation of a Soluble Human Programmed Death-1 Protein Assay.

    Science.gov (United States)

    Ni, Yan G; Yuan, Xiling; Newitt, John A; Peterson, Jon E; Gleason, Carol R; Haulenbeek, Jonathan; Santockyte, Rasa; Lafont, Virginie; Marsilio, Frank; Neely, Robert J; DeSilva, Binodh; Piccoli, Steven P

    2015-07-01

    Programmed death-1 (PD-1) protein is a co-inhibitory receptor which negatively regulates immune cell activation and permits tumors to evade normal immune defense. Anti-PD-1 antibodies have been shown to restore immune cell activation and effector function-an exciting breakthrough in cancer immunotherapy. Recent reports have documented a soluble form of PD-1 (sPD-1) in the circulation of normal and disease state individuals. A clinical assay to quantify sPD-1 would contribute to the understanding of sPD-1-function and facilitate the development of anti-PD-1 drugs. Here, we report the development and validation of a sPD-1 protein assay. The assay validation followed the framework for full validation of a biotherapeutic pharmacokinetic assay. A purified recombinant human PD-1 protein was characterized extensively and was identified as the assay reference material which mimics the endogenous analyte in structure and function. The lower limit of quantitation (LLOQ) was determined to be 100 pg/mL, with a dynamic range spanning three logs to 10,000 pg/mL. The intra- and inter-assay imprecision were ≤15%, and the assay bias (percent deviation) was ≤10%. Potential matrix effects were investigated in sera from both normal healthy volunteers and selected cancer patients. Bulk-prepared frozen standards and pre-coated Streptavidin plates were used in the assay to ensure consistency in assay performance over time. This assay appears to specifically measure total sPD-1 protein since the human anti-PD-1 antibody, nivolumab, and the endogenous ligands of PD-1 protein, PDL-1 and PDL-2, do not interfere with the assay.

  4. Soluble Mesothelin-Related Protein in Malignant Pleural Mesothelioma

    International Nuclear Information System (INIS)

    AZIM, H.A.; GAAFAR, R.; KHORSHID, O.; ABDEL SALAM, I.; ASHMAWY, A.; EL-GUINDY, S.; ELATTAR, I.

    2008-01-01

    Background and Purpose: Building-up evidence suggests that soluble mesothelinrelated protein (SMRP) carries a diagnostic and a prognostic value in malignant pleural mesothelioma (MPM). Egypt suffers endemic asbestosis and thus this study was conducted to evaluate the sensitivity and specificity of SMRP in patients with MPM and to correlate this marker with known clinico pathological prognostic factors. Material and Methods: During the period from January 2006 till March 2008, Serum samples were obtained from MPM patients presenting to the Egyptian National Cancer Institute, Cairo University. Serum samples were provided from patients with breast cancer and from healthy individuals to function as controls. The SMRP was assayed using the ELISA technique and correlations were made with different clinico-pathological prognostic parameters. Results: 83 patients (50 MPM and 33 breast cancer) as well as 22 healthy individuals were enrolled in this study. Serum SMRP levels were not different between patients with breast cancer and healthy controls (p>0.05). However, there was a significant difference between MPM patients and the other two groups (p<0.0001). ROC analysis showed an AUC=0.765 for differentiating between the controls and MPM with a best statistical cut-off of 7.22 nM/L (sensitivity=66%, specificity=70.9%). The mean SMRP concentrations were significantly higher in patients with advanced disease (p=0.038), poor performance status (p=0.017) and high alkaline phosphatase (p=0.015). The mean SMRP concentrations were also higher in males, elderly patients, asbestos-exposed patients, epithelioid subtypes and patients with high platelet and leucocytic counts. However, these differences did not reach statistical significance. 224 Conclusions: This study confirms that SMRP is of considerable sensitivity and specificity in Egyptian patients with MPM. Higher levels are frequently seen in patients with high tumor burden, which could be helpful in monitoring response to

  5. Complement activating soluble pattern recognition molecules with collagen-like regions, mannan-binding lectin, ficolins and associated proteins

    DEFF Research Database (Denmark)

    Thiel, Steffen

    2007-01-01

    Mannan-binding lectin (MBL), L-ficolin, M-ficolin and H-ficolin are all complement activating soluble pattern recognition molecules with recognition domains linked to collagen-like regions. All four may form complexes with four structurally related proteins, the three MBL-associated serine...... proteases (MASPs), MASP-1, MASP-2 and MASP-3, and a smaller MBL-associated protein (MAp19). The four recognition molecules recognize patterns of carbohydrate or acetyl-group containing ligands. After binding to the relevant targets all four are able to activate the complement system. We thus have a system...... where four different and/or overlapping patterns of microbial origin or patterns of altered-self may be recognized, but in all cases the signalling molecules, the MASPs, are shared. MASP-1 and MASP-3 are formed from one gene, MASP1/3, by alternative splicing generating two different mRNAs from a single...

  6. Changes in the protein profile of Habanero pepper (Capsicum ...

    African Journals Online (AJOL)

    2012-06-12

    Jun 12, 2012 ... (Capsicum chinense J.) somatic embryos during ... molecular weights to those reported for storage proteins in other .... were isolated by cutting with a razor blade. ... electrophoresis (SDS-PAGE; 5% stacking gel [pH 6.8], 15% running ... Developmental stages correspond to C) globular, D) heart-shaped, ...

  7. Most Massive Globular Cluster in Our Galaxy

    Science.gov (United States)

    1994-05-01

    Far down in the southern sky, in the constellation of Centaurus, a diffuse spot of light can be perceived with the unaided eye. It may be unimpressive, but when seen through a telescope, it turns out to be a beautiful, dense cluster of innumerable stars [1]. Omega Centauri, as this object is called, is the brightest of its type in the sky. We refer to it as a "globular cluster", due to its symmetric form. It belongs to our Milky Way galaxy and astrophysical investigations have shown that it is located at a distance of about 16,500 light-years (1 light-year = 9,460,000,000,000 km). Nobody knows for sure how many individual stars it contains, but recent estimates run into the millions. Most of these stars are more than 10,000 million years old and it is generally agreed that Omega Centauri has a similar age. Measurements of its motion indicate that Omega Centauri plows through the Milky Way in an elongated orbit. It is not easy to understand how it has managed to keep its stars together during such an extended period. MEASURING STELLAR VELOCITIES IN OMEGA CENTAURI A group of astronomers [2] have recently carried through a major investigation of Omega Centauri. After many nights of observations at the ESO La Silla observatory, they now conclude that not only is this globular cluster the brightest, it is indeed by far the most massive known in the Milky Way. The very time-consuming observations were made during numerous observing sessions over a period of no less than 13 years (1981-1993), with the photoelectric spectrometer CORAVEL mounted on the 1.5-m Danish telescope at La Silla. The CORAVEL instrument (COrelation RAdial VELocities) was built in a joint effort between the Geneva (Switzerland) and Marseilles (France) observatories. It functions according to the cross-correlation technique, by means of which the spectrum of the observed star is compared with a "standard stellar spectrum" [3]. HOW HEAVY IS OMEGA CENTAURI? In the present study, a total of 1701

  8. BVRI main-sequence photometry of the globular cluster M4

    International Nuclear Information System (INIS)

    Alcaino, G.; Liller, W.

    1984-01-01

    We present BV and RI photographic photometry of 1421 and 189 stars, respectively, in the intermediate metallicity globular cluster M4 (NGC 6121). This investigation includes the first results of RI main-sequence photometry of a globular cluster. The use of longer wavelengths and longer color baselines provides the potential of improved isochrone fittings and underscores the urgent need for calculations of RI synthetic isochrones to be compared with observations. The Pickering-Racine wedge was used with the ESO 3.6 m telescope, the Las Campanas 2.5 m du Pont telescope, and the CTIO 1 m Yale telescope to extend the photoelectric limit from Vroughly-equal16.1 to Vroughly-equal19.1. We have determined the position of the main-sequence turnoff to lie at V = 16.6 +- 0.2 (m.e.) and B-V = 0.80 +- 0.03 (m.e.). A comparison of our BV observations with the CCD data of Richer and Fahlman shows excellent agreement: the two fifucial main sequences agree at all points to within 0.025 mag and, on average, to 0.013 mag. For the cluster we derive a distance modulus (m-M)/sub V/ = 12.52 +- 0.2 and reddening E(B-V) = 0.44 +- 0.03, results which confirm that at a distance of 2 kpc, M4 is the closest globular clusters to the Sun. Using the isochrones of VandenBerg, we deduce an age 13 +- 2 Gyr. As noted in several other investigations, there is a striking deficiency of stars in certain parts of the color-magnitude diagram; in M4 we find a pronounced gap over approx.0.6 mag at the base of the subgiant branch

  9. Use of plasma C-reactive protein, procalcitonin, neutrophils,macrophage migration inhibitory factor, soluble urokinase-type plasminogen activator receptor, and soluble triggering receptor expressed on myeloid cells-1 in combination to diagnose infections: a prospective study

    DEFF Research Database (Denmark)

    Kofoed, Kristian; Andersen, Ove; Kronborg, Gitte

    2007-01-01

    the diagnostic characteristics of novel and routinely used biomarkers of sepsis alone and in combination. Methods: This prospective cohort study included patients with systemic inflammatory response syndrome who were suspected of having community-acquired infections. It was conducted in a medical emergency...... department and department of infectious diseases at a university hospital. A multiplex immunoassay measuring soluble urokinase-type plasminogen activator (suPAR) and soluble triggering receptor expressed on myeloid cells (sTREM)-1 and macrophage migration inhibitory factor (MIF) was used in parallel...... with standard measurements of C-reactive protein (CRP), procalcitonin (PCT), and neutrophils. Two composite markers were constructed – one including a linear combination of the three best performing markers and another including all six – and the area under the receiver operating characteristic curve (AUC...

  10. Water Dynamics in Protein Hydration Shells: The Molecular Origins of the Dynamical Perturbation

    Science.gov (United States)

    2014-01-01

    Protein hydration shell dynamics play an important role in biochemical processes including protein folding, enzyme function, and molecular recognition. We present here a comparison of the reorientation dynamics of individual water molecules within the hydration shell of a series of globular proteins: acetylcholinesterase, subtilisin Carlsberg, lysozyme, and ubiquitin. Molecular dynamics simulations and analytical models are used to access site-resolved information on hydration shell dynamics and to elucidate the molecular origins of the dynamical perturbation of hydration shell water relative to bulk water. We show that all four proteins have very similar hydration shell dynamics, despite their wide range of sizes and functions, and differing secondary structures. We demonstrate that this arises from the similar local surface topology and surface chemical composition of the four proteins, and that such local factors alone are sufficient to rationalize the hydration shell dynamics. We propose that these conclusions can be generalized to a wide range of globular proteins. We also show that protein conformational fluctuations induce a dynamical heterogeneity within the hydration layer. We finally address the effect of confinement on hydration shell dynamics via a site-resolved analysis and connect our results to experiments via the calculation of two-dimensional infrared spectra. PMID:24479585

  11. Biophysics of protein evolution and evolutionary protein biophysics

    Science.gov (United States)

    Sikosek, Tobias; Chan, Hue Sun

    2014-01-01

    The study of molecular evolution at the level of protein-coding genes often entails comparing large datasets of sequences to infer their evolutionary relationships. Despite the importance of a protein's structure and conformational dynamics to its function and thus its fitness, common phylogenetic methods embody minimal biophysical knowledge of proteins. To underscore the biophysical constraints on natural selection, we survey effects of protein mutations, highlighting the physical basis for marginal stability of natural globular proteins and how requirement for kinetic stability and avoidance of misfolding and misinteractions might have affected protein evolution. The biophysical underpinnings of these effects have been addressed by models with an explicit coarse-grained spatial representation of the polypeptide chain. Sequence–structure mappings based on such models are powerful conceptual tools that rationalize mutational robustness, evolvability, epistasis, promiscuous function performed by ‘hidden’ conformational states, resolution of adaptive conflicts and conformational switches in the evolution from one protein fold to another. Recently, protein biophysics has been applied to derive more accurate evolutionary accounts of sequence data. Methods have also been developed to exploit sequence-based evolutionary information to predict biophysical behaviours of proteins. The success of these approaches demonstrates a deep synergy between the fields of protein biophysics and protein evolution. PMID:25165599

  12. Karakteristik Protein dan Nitrogen Non Protein Daging Ikan Cucut Lanyam (Charcharhinus limbatus (Characteristics of Protein and Non Protein Nitrogen in Lanyam Shark Muscle

    Directory of Open Access Journals (Sweden)

    Yuspihana Fitrial

    2017-02-01

    Based on protein solubility of Lanyam muscle at pH 1.5 to 12 obtained two points which is minimum solubility at pH 4.5 and pH 9. Based on the classification Osborn, Lanyam muscle contained albumin (28.64%, globulin (13:44%, prolamin (03.29%, glutelin (33.70%. Observation of non-protein nitrogen levels indicated that the washing process was very effective to reduce non-protein nitrogen levels up to 62.34% and urea levels up to 58% . Differential Scanning Calorimetry Study of Lanyam mince showed two types of protein that has a different stability to heat and after added 2.5% NaCl formed a peak which is a fusion of both these proteins

  13. Optimized sample preparation for two-dimensional gel electrophoresis of soluble proteins from chicken bursa of Fabricius

    Directory of Open Access Journals (Sweden)

    Zheng Xiaojuan

    2009-10-01

    Full Text Available Abstract Background Two-dimensional gel electrophoresis (2-DE is a powerful method to study protein expression and function in living organisms and diseases. This technique, however, has not been applied to avian bursa of Fabricius (BF, a central immune organ. Here, optimized 2-DE sample preparation methodologies were constructed for the chicken BF tissue. Using the optimized protocol, we performed further 2-DE analysis on a soluble protein extract from the BF of chickens infected with virulent avibirnavirus. To demonstrate the quality of the extracted proteins, several differentially expressed protein spots selected were cut from 2-DE gels and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS. Results An extraction buffer containing 7 M urea, 2 M thiourea, 2% (w/v 3-[(3-cholamidopropyl-dimethylammonio]-1-propanesulfonate (CHAPS, 50 mM dithiothreitol (DTT, 0.2% Bio-Lyte 3/10, 1 mM phenylmethylsulfonyl fluoride (PMSF, 20 U/ml Deoxyribonuclease I (DNase I, and 0.25 mg/ml Ribonuclease A (RNase A, combined with sonication and vortex, yielded the best 2-DE data. Relative to non-frozen immobilized pH gradient (IPG strips, frozen IPG strips did not result in significant changes in the 2-DE patterns after isoelectric focusing (IEF. When the optimized protocol was used to analyze the spleen and thymus, as well as avibirnavirus-infected bursa, high quality 2-DE protein expression profiles were obtained. 2-DE maps of BF of chickens infected with virulent avibirnavirus were visibly different and many differentially expressed proteins were found. Conclusion These results showed that method C, in concert extraction buffer IV, was the most favorable for preparing samples for IEF and subsequent protein separation and yielded the best quality 2-DE patterns. The optimized protocol is a useful sample preparation method for comparative proteomics analysis of chicken BF tissues.

  14. Radiation synthesis of a water-soluble temperature sensitive polymer, activated copolymer and applications in immobilization of proteins

    International Nuclear Information System (INIS)

    Zhai Maolin; Ha Hongfei; Wu Jilan

    1993-01-01

    In this work the radiation polymerization of N-isopropylacrylamide (NIPAAM) in aqueous solutions has been carried out and a water-soluble, temperature sensitive polymer and copolymer were obtained by using γ-rays from Co-60 source at room temperature. We have gained the optimum dose and dose-rate of radiation synthesis of linear polyNIPAAM through determining conversion yield and viscosity. In order to immobilize protein (BSA) and enzyme (HRP) into this water-soluble polymer, we prepared an activated copolymer, poly(N-isopropylacrylamide-co-N-acryloxysuccinimide). The BSA and HRP has been immobilized onto the activated copolymer. The BSA (HRP)/copolymer conjugates still kept the original thermally sensitive properties of the linear polyNIPAAM. The conjugation yield of BSA to the activated copolymer decreased with increasing dose. Immobilized HRP was stable at 0 o C for a long time and has, at least, 4 days stability at room temperature. Immobilized HRP activity was lowered when the temperature was raised. This phenomenon was reversible and the immobilized HRP regained activity. The optimum pH of the immobilized HRP shifted from ca.5 upward to ca. 7. (author)

  15. In silico local structure approach: a case study on outer membrane proteins.

    Science.gov (United States)

    Martin, Juliette; de Brevern, Alexandre G; Camproux, Anne-Claude

    2008-04-01

    The detection of Outer Membrane Proteins (OMP) in whole genomes is an actual question, their sequence characteristics have thus been intensively studied. This class of protein displays a common beta-barrel architecture, formed by adjacent antiparallel strands. However, due to the lack of available structures, few structural studies have been made on this class of proteins. Here we propose a novel OMP local structure investigation, based on a structural alphabet approach, i.e., the decomposition of 3D structures using a library of four-residue protein fragments. The optimal decomposition of structures using hidden Markov model results in a specific structural alphabet of 20 fragments, six of them dedicated to the decomposition of beta-strands. This optimal alphabet, called SA20-OMP, is analyzed in details, in terms of local structures and transitions between fragments. It highlights a particular and strong organization of beta-strands as series of regular canonical structural fragments. The comparison with alphabets learned on globular structures indicates that the internal organization of OMP structures is more constrained than in globular structures. The analysis of OMP structures using SA20-OMP reveals some recurrent structural patterns. The preferred location of fragments in the distinct regions of the membrane is investigated. The study of pairwise specificity of fragments reveals that some contacts between structural fragments in beta-sheets are clearly favored whereas others are avoided. This contact specificity is stronger in OMP than in globular structures. Moreover, SA20-OMP also captured sequential information. This can be integrated in a scoring function for structural model ranking with very promising results. (c) 2007 Wiley-Liss, Inc.

  16. Characterization of dry globular proteins and protein fibrils by synchrotron radiation vacuum UV circular dichroism

    DEFF Research Database (Denmark)

    Nesgaard, Lise W.; Hoffmann, Søren Vrønning; Andersen, Christian Beyschau

    2008-01-01

    Circular dichroism using synchrotron radiation (SRCD) can extend the spectral range down to approximately 130 nm for dry proteins, potentially providing new structural information. Using a selection of dried model proteins, including alpha-helical, beta-sheet, and mixed-structure proteins, we...... with previously published theoretical calculations related to pi-orbital transitions. We also show that drying does not lead to large changes in the secondary structure and does not induce orientational artifacts. In combination with principal component analysis, our SRCD data allow us to distinguish between two...... different types of protein fibrils, highlighting that bona fide fibrils formed by lysozyme are structurally more similar to the nonclassical fibrillar aggregates formed by the SerADan peptide than with the amyloid formed by alpha-synuclein. Thus, despite the lack of direct structural conclusions...

  17. A period-luminosity relation for Mira variables in globular clusters and its impact on the distance scale

    International Nuclear Information System (INIS)

    Menzies, J.W.; Whitelock, P.A.

    1985-01-01

    JHKL photometry is presented for 31 red variables in 15 galactic globular clusters. The photometry of the Mira variables is used to find absolute bolometric magnitudes and an Msub(bol)-log P relation which differs from the one found for LMC Miras. This can be understood only if there is some systematic error in the globular cluster and/or LMC distance scales or if there is some fundamental difference between the cluster Miras and those in the LMC. (author)

  18. The nutritive value of condensed wheat distillers solubles for cattle

    NARCIS (Netherlands)

    Boever, De J.L.; Blok, M.C.; Millet, S.; Vanacker, J.; Campeneere, De S.

    2016-01-01

    The chemical composition and the energy and protein value of five batches of condensed distillers solubles (CDS) originating from wheat were determined. The net energy for lactation (NEL) was derived from digestion coefficients obtained with sheep. The true protein digested in the small intestine

  19. Electrorheological properties of suspensions of hollow globular titanium oxide/polypyrrole particles

    Czech Academy of Sciences Publication Activity Database

    Sedlačík, M.; Mrlík, M.; Pavlínek, V.; Sáha, P.; Quadrat, Otakar

    2012-01-01

    Roč. 290, č. 1 (2012), s. 41-48 ISSN 0303-402X R&D Projects: GA ČR GA202/09/1626 Institutional research plan: CEZ:AV0Z40500505 Keywords : electrorheology * titanium oxide * hollow globular clusters Subject RIV: JI - Composite Materials Impact factor: 2.161, year: 2012

  20. Altering protein surface charge with chemical modification modulates protein–gold nanoparticle aggregation

    International Nuclear Information System (INIS)

    Jamison, Jennifer A.; Bryant, Erika L.; Kadali, Shyam B.; Wong, Michael S.; Colvin, Vicki L.; Matthews, Kathleen S.; Calabretta, Michelle K.

    2011-01-01

    Gold nanoparticles (AuNP) can interact with a wide range of molecules including proteins. Whereas significant attention has focused on modifying the nanoparticle surface to regulate protein–AuNP assembly or influence the formation of the protein “corona,” modification of the protein surface as a mechanism to modulate protein–AuNP interaction has been less explored. Here, we examine this possibility utilizing three small globular proteins—lysozyme with high isoelectric point (pI) and established interactions with AuNP; α-lactalbumin with similar tertiary fold to lysozyme but low pI; and myoglobin with a different globular fold and an intermediate pI. We first chemically modified these proteins to alter their charged surface functionalities, and thereby shift protein pI, and then applied multiple methods to assess protein–AuNP assembly. At pH values lower than the anticipated pI of the modified protein, AuNP exposure elicits changes in the optical absorbance of the protein–NP solutions and other properties due to aggregate formation. Above the expected pI, however, protein–AuNP interaction is minimal, and both components remain isolated, presumably because both species are negatively charged. These data demonstrate that protein modification provides a powerful tool for modulating whether nanoparticle–protein interactions result in material aggregation. The results also underscore that naturally occurring protein modifications found in vivo may be critical in defining nanoparticle–protein corona compositions.

  1. Escherichia coli fusion carrier proteins act as solubilizing agents for recombinant uncoupling protein 1 through interactions with GroEL

    International Nuclear Information System (INIS)

    Douette, Pierre; Navet, Rachel; Gerkens, Pascal; Galleni, Moreno; Levy, Daniel; Sluse, Francis E.

    2005-01-01

    Fusing recombinant proteins to highly soluble partners is frequently used to prevent aggregation of recombinant proteins in Escherichia coli. Moreover, co-overexpression of prokaryotic chaperones can increase the amount of properly folded recombinant proteins. To understand the solubility enhancement of fusion proteins, we designed two recombinant proteins composed of uncoupling protein 1 (UCP1), a mitochondrial membrane protein, in fusion with MBP or NusA. We were able to express soluble forms of MBP-UCP1 and NusA-UCP1 despite the high hydrophobicity of UCP1. Furthermore, the yield of soluble fusion proteins depended on co-overexpression of GroEL that catalyzes folding of polypeptides. MBP-UCP1 was expressed in the form of a non-covalent complex with GroEL. MBP-UCP1/GroEL was purified and characterized by dynamic light scattering, gel filtration, and electron microscopy. Our findings suggest that MBP and NusA act as solubilizing agents by forcing the recombinant protein to pass through the bacterial chaperone pathway in the context of fusion protein

  2. Soluble haemoglobin is a marker of recent Plasmodium falciparum infections

    DEFF Research Database (Denmark)

    Jakobsen, P H; Bygbjerg, I C; Theander, T G

    1997-01-01

    Monoclonal antibodies (Mab) were raised against haemoglobin (Hb) associated with Plasmodium falciparum protein and used to develop an ELISA, measuring circulating levels of released Hb. This assay was evaluated in different malaria patients in parallel with ELISA assays for C-reactive protein (CR...... after treatment. Soluble Hb levels were associated with malariometric parameters in a similar fashion to haptoglobin. The new Mab-based assay for measuring soluble Hb in the peripheral blood of malaria patients may be useful for future epidemiological studies of malaria....

  3. Functional properties of dioscorin, a soluble viscous protein from Japanese yam (Dioscorea opposita thunb.) tuber mucilage Tororo.

    Science.gov (United States)

    Nagai, Takeshi; Nagashima, Toshio

    2006-01-01

    A soluble viscous protein was purified from yam (Dioscorea opposita Thunb.) tuber mucilage tororo by chromatographic steps, and its functional properties were estimated. The purified dioscorin having the molecular weight of about 200 kDa exhibited high scavenging activities against hydroxyl radicals (IC50 = 195.1 microg/ml) and superoxide anion radicals (IC50 = 92.7 microg/ml). Moreover, it showed extremely high angiotensin I-converting enzyme inhibitory activity (IC50 = 41.1 microg/ml). The results suggested that yam D. opposita tuber has a wide spectrum of strong antioxidative and antihypertensive activities and it could be utilized as a source of natural antioxidant.

  4. Extracting Tenebrio molitor protein while preventing browning: effect of pH and NaCl on protein yield

    NARCIS (Netherlands)

    Yi, L.; Boekel, van T.; Lakemond, C.M.M.

    2017-01-01

    The potential of insects as an alternative protein source for food applications was investigated by studying the effect of pH and NaCl on extraction yield of water-soluble proteins from Tenebrio molitor, while preventing browning due to polyphenol oxidation. Minimum protein solubility (29.6%) was at

  5. Sub-nanoscale surface ruggedness provides a water-tight seal for exposed regions in soluble protein structure.

    Directory of Open Access Journals (Sweden)

    Erica Schulz

    2010-09-01

    Full Text Available Soluble proteins must maintain backbone hydrogen bonds (BHBs water-tight to ensure structural integrity. This protection is often achieved by burying the BHBs or wrapping them through intermolecular associations. On the other hand, water has low coordination resilience, with loss of hydrogen-bonding partnerships carrying significant thermodynamic cost. Thus, a core problem in structural biology is whether natural design actually exploits the water coordination stiffness to seal the backbone in regions that are exposed to the solvent. This work explores the molecular design features that make this type of seal operative, focusing on the side-chain arrangements that shield the protein backbone. We show that an efficient sealing is achieved by adapting the sub-nanoscale surface topography to the stringency of water coordination: an exposed BHB may be kept dry if the local concave curvature is small enough to impede formation of the coordination shell of a penetrating water molecule. Examination of an exhaustive database of uncomplexed proteins reveals that exposed BHBs invariably occur within such sub-nanoscale cavities in native folds, while this level of local ruggedness is absent in other regions. By contrast, BHB exposure in misfolded proteins occurs with larger local curvature promoting backbone hydration and consequently, structure disruption. These findings unravel physical constraints fitting a spatially dependent least-action for water coordination, introduce a molecular design concept, and herald the advent of water-tight peptide-based materials with sufficient backbone exposure to remain flexible.

  6. Tracing dust in old stellar populations : the mid-infrared spectrum of globular cluster AGB stars

    International Nuclear Information System (INIS)

    Richter, H.

    2010-01-01

    Asymptotic Giant Branch (AGB) stars are considered to be the main stellar dust producers in the universe. Their dusty circumstellar shells leave fingerprints in the mid-infrared (MIR) spectra of AGB stars and in unresolved old stellar populations. Bressan et al. (2007) showed that co-added MIR-spectra of AGB stars of known luminosity, metallicity and age (like those found in the Galactic globular cluster NGC104) can be used to model the dust excess in early-type galaxies. This work aims to improve our understanding of the MIR-spectra of old stellar populations with respect to their metallicities by studying a large sample of AGB stars in Galactic globular clusters. A sample of AGB stars (taken from Lebzelter et al. 2006 and Sloan et al. 2010) is used to produce co-added MIR-spectra of globular cluster combinations for three metallicity groups. Each group consists of several globular clusters with similar age and metallicity. Combining the clusters leads to a higher number of AGB stars with available Spitzer spectra in each group. The low metallicity group (Z=0.0038) consists of five globular clusters with 18 AGB star spectra, the intermediate (Z=0.0058) and high (Z=0.01) metallicity groups both include three clusters with eight and seven available MIR-spectra, respectively. Stars within the 90% mass radius of each globular cluster are used to generate 2MASS Color- Magnitude diagrams (CMDs) of each cluster combination. Binning the stars in the CMDs with respect to their MK-values results in Luminosity Functions (LFs) for the cluster combinations. The LFs based on 2MASS data are compared to LFs obtained using theoretical isochrones from the Padova group (Bertelli et al. 2008, Marigo et al. 2008). Using the 2MASS LFs integrated MIR-spectra of the three globular cluster combinations are derived by weighting the existing spectra with the total number of AGB stars within each MK-bin of the LFs along the upper giant branch. This relies on the assumption that stars that

  7. Intraperitoneal administration of the globular adiponectin gene ameliorates diabetic nephropathy in Wistar rats.

    Science.gov (United States)

    Yuan, Fang; Liu, Ying-Hong; Liu, Fu-You; Peng, You-Ming; Tian, Jun-Wei

    2014-06-01

    The present study investigated the potential effects of the long-term expression of exogenous adiponectin (ADPN) on normal and diabetic kidneys. Type 2 diabetes mellitus models were induced by high-lipid and high-sucrose feeding plus intraperitoneal injection of streptozotocin. The recombinant plasmid pIRES2-EGFP-gAd, which is able to co-express globular ADPN (gAd) and enhanced green fluorescent protein (EGFP), was intraperitoneally injected into rat models mediated by Lipofectamine. In total, 32 Wistar rats were randomly assigned into four groups: the normal control group, the diabetes group, the diabetes group treated with pIRES2-EGFP-gAd and the diabetes group treated with pIRES2-EGFP. After 12 weeks, serum biochemistry and urine albumin levels were measured. The kidneys were collected to assess the generation of reactive oxygen species (ROS) and the renal pathological changes were observed by light microcopy. The protein expression of endothelial nitric oxide synthase (eNOS), transforming growth factor-β1 (TGF-β1) and phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK) were determined by an immunohistochemical staining method and western blot analysis. Intraperitoneal injection of the human gAd gene via Lipofectamine resulted in abundant ADPN protein in the kidney. In the diabetic rats, the delivery of the exogenous gAd gene ameliorated the progression of diabetic nephropathy (DN). ADPN attenuated urine albumin excretion in the diabetic rats. ADPN also mitigated glomerular mesangial expansion, reduced the generation of ROS and prevented interstitial fibrosis. In addition, the expression of gAd inhibited the renal expression of TGF-β1, promoted the protein expression of eNOS and activated the opening of the AMPK signaling pathway in the renal tissues of the diabetic rats. Despite the effects of ADPN on DN being controversial, these observations indicate that the supplementation of ADPN is beneficial in ameliorating DN in rats.

  8. Spider Silk Fibers Spun from Soluble Recombinant Silk Produced in Mammalian Cells

    Science.gov (United States)

    Lazaris, Anthoula; Arcidiacono, Steven; Huang, Yue; Zhou, Jiang-Feng; Duguay, François; Chretien, Nathalie; Welsh, Elizabeth A.; Soares, Jason W.; Karatzas, Costas N.

    2002-01-01

    Spider silks are protein-based ``biopolymer'' filaments or threads secreted by specialized epithelial cells as concentrated soluble precursors of highly repetitive primary sequences. Spider dragline silk is a flexible, lightweight fiber of extraordinary strength and toughness comparable to that of synthetic high-performance fibers. We sought to ``biomimic'' the process of spider silk production by expressing in mammalian cells the dragline silk genes (ADF-3/MaSpII and MaSpI) of two spider species. We produced soluble recombinant (rc)-dragline silk proteins with molecular masses of 60 to 140 kilodaltons. We demonstrated the wet spinning of silk monofilaments spun from a concentrated aqueous solution of soluble rc-spider silk protein (ADF-3; 60 kilodaltons) under modest shear and coagulation conditions. The spun fibers were water insoluble with a fine diameter (10 to 40 micrometers) and exhibited toughness and modulus values comparable to those of native dragline silks but with lower tenacity. Dope solutions with rc-silk protein concentrations >20% and postspinning draw were necessary to achieve improved mechanical properties of the spun fibers. Fiber properties correlated with finer fiber diameter and increased birefringence.

  9. Tyrosine phosphorylation of a 66KD soluble protein and augmentation of lectin induced mitogenesis by DMSO in human T lymphocytes

    International Nuclear Information System (INIS)

    Wedner, H.J.; Bass, G.

    1986-01-01

    The authors have demonstrated that induction of mitogenesis in human T lymphocytes is associated with the tyrosine phosphorylation of a 66KD soluble substrate-TPP 66. Since DMSO has been shown to be a non-specific stimulator of tyrosine protein kinases they have examined the effect of DMSO on both activation and tyrosine phosphorylation in human T cells. Human peripheral blood T lymphocytes were isolated by dextran sedimentation, Ficol/Paque centrifugation and nylon wool filtration. Phosphorylation was performed in cells incubated with [ 32 P] orthophosphate followed by DMSO for 30 min. TPP 66 was identified by 2-D PAGE, autoradiography, and HV electrophoresis of the hydrolyzed protein. Concentrations of DMSO from 1% to 50% induced the tyrosine phosphorylation of TPP 66 with maximal stimulation seen at 20%. DMSO alone did not activate the T cells (measured by [ 3 H] thymidine incorporation) when tested at high concentrations for 30 sec to 10 min. (longer incubations were markedly toxic) or low concentrations for 12 to 48 hrs. Low concentrations of DMSO 0.1%-0.5% did however, markedly augment [ 3 H] thymidine incorporation induced by PHA or Con A. These data suggest that tyrosine phosphorylation of TPP 66 alone may not constitute sufficient signal for the activation sequence to begin but the phosphorylation of this soluble substrate may be a critical factor in the propagation of the activation sequence

  10. Cold-set globular protein gels: Interactions, structure and rheology as a function of protein concentration.

    NARCIS (Netherlands)

    Alting, A.C.; Hamer, R.J.; Kruif, de C.G.

    2003-01-01

    We identified the contribution of covalent and noncovalent interactions to the scaling behavior of the structural and rheological properties in a cold gelling protein system. The system we studied consisted of two types of whey protein aggregates, equal in size but different in the amount of

  11. Separate introns gained within short and long soluble peridinin-chlorophyll a-protein genes during radiation of Symbiodinium (Dinophyceae) clade A and B lineages - PLoS One

    Science.gov (United States)

    Here we document introns in two Symbiodinium clades that were most likely gained following divergence of this genus from other peridinin-containing dinoflagellate lineages. Soluble peridinin-chlorophyll a-proteins (sPCP) occur in short and long forms in different species, and all...

  12. Recent developments in the theory of protein folding: searching for the global energy minimum.

    Science.gov (United States)

    Scheraga, H A

    1996-04-16

    Statistical mechanical theories and computer simulation are being used to gain an understanding of the fundamental features of protein folding. A major obstacle in the computation of protein structures is the multiple-minima problem arising from the existence of many local minima in the multidimensional energy landscape of the protein. This problem has been surmounted for small open-chain and cyclic peptides, and for regular-repeating sequences of models of fibrous proteins. Progress is being made in resolving this problem for globular proteins.

  13. Intrachromosomal exchange aberrations predicted on the basis of globular interphase chromosome model

    International Nuclear Information System (INIS)

    Andreev, S.G.; Eidelman, Yu.A.

    2002-01-01

    One of the key questions in understanding mechanisms of chromosome aberration production is how does interphase chromosome structure affect aberration formation. To explore this a modelling approach is presented which combines Monte Carlo simulation of both a particle track and interphase chromosome structure. The structural state of interphase chromosome influences a dose-effect relationship for intrachromosomal exchange aberrations (intrachanges). It is shown that intrachanges are induced frequently by both X rays and a particles if the chromosome is in the condensed globular but not in the decondensed coiled state. Truly simple intra-arm intrachanges induced by X rays are dose squared in coiled chromosomes, but exhibit linear dose dependence in globular chromosomes. Experimental data on interarm intrachanges obtained by dual arm chromosome painting are analysed by means of the technique presented. Results of analysis support the conclusion about the arms proximity of chromosome 1 in human lymphocytes. (author)

  14. Properties of the disk system of globular clusters

    International Nuclear Information System (INIS)

    Armandroff, T.E.

    1989-01-01

    A large refined data sample is used to study the properties and origin of the disk system of globular clusters. A scale height for the disk cluster system of 800-1500 pc is found which is consistent with scale-height determinations for samples of field stars identified with the Galactic thick disk. A rotational velocity of 193 + or - 29 km/s and a line-of-sight velocity dispersion of 59 + or - 14 km/s have been found for the metal-rich clusters. 70 references

  15. Chemical Abundances of Red Giant Branch Stars in the Globular Clusters NGC 6333 and NGC 6366

    Science.gov (United States)

    Johnson, Christian I.; Rich, R. M.; Pilachowski, C. A.; Kunder, A. M.

    2013-01-01

    We present chemical abundances and radial velocities for >20 red giant branch (RGB) stars in the Galactic globular clusters NGC 6333 ([Fe/H]≈-1.8) and NGC 6366 ([Fe/H]≈-0.6). The results are based on moderate resolution (R=18,000), high signal-to-noise ratio (>100) spectra obtained with the Hydra multifiber positioner and bench spectrograph on the WIYN 3.5m telescope at Kitt Peak National Observatory. Both objects are likely associated with the Galactic bulge globular cluster system, and we therefore compare the cluster abundance patterns with those of nearby bulge field stars. Additionally, we investigate differences in the O-Na anticorrelation and neutron-capture element dispersion between the two clusters, and compare their abundance patterns with those of similar metallicity halo globular clusters. This material is based upon work supported by the National Science Foundation under award No. AST-1003201 to C.I.J. C.A.P. gratefully acknowledges support from the Daniel Kirkwood Research Fund at Indiana University. R.M.R. acknowledges support from NSF grant AST-0709479 and AST-121120995.

  16. PROTEIN FRACTIONS AND IN VITRO FERMENTATION OF PROTEIN FEEDS FOR RUMINANTS

    Directory of Open Access Journals (Sweden)

    Angel L. Guevara-Mesa

    2011-05-01

    Full Text Available The objective of this study was to evaluate 20 protein feeds grouped in forages, vegetal by- products and animal by-products used for ruminant diets. Protein fractions (PF: A, non-protein nitrogen (NPN; B1, buffer-soluble protein; B2, buffer-insoluble, NDF-soluble protein; B3, NDF-insoluble, ADF-soluble protein; and C, ADF-insoluble protein, were determined for each ingredient.  Protein composition was correlated with total gas production in vitro (GP, gas production rate (S, lag time (L, DM disappearance (DMDIV and residual protein (RPIV. The completely randomised designed was analysed using mixed proc. and Tukey contrasts. Forages contained 18.29, 7.86, 66.00, 2.96, 4.89% of fractions A, B1, B2, B3 and C, respectively. Vegetable by-products contained 22.55, 4.55, 59.51, 8.84, 4.55% of each fraction, in the same order. Animal by-products contained 19.13, 4.52, 70.24, 3.74, 2.37% of each fraction, in the same order. Vetch, wheat bran and poultry litter had the greatest Vmax in each group. Vmax was correlated (P≤0.01 with total protein (r = -0.45, ADF (r = 0.27 and DMDIV (r = 0.61. In conclusion, there were differences in protein composition and kinetics of in vitro gas production among ingredients.

  17. Study on REE bound water-soluble polysaccharides in plant

    International Nuclear Information System (INIS)

    Wang Yuqi; Guo Fanqing; Xu Lei; Chen Hongmin; Sun Jingxin; Cao Guoyin

    1999-01-01

    The binding of REE with water-soluble polysaccharides (PSs) in leaves of fern Dicranopteris Dichotoma (DD) has been studied by molecular activation analysis. The cold-water-soluble and hot-water-soluble PSs in leaves of DD were obtained by using biochemical separation techniques. The PSs of non-deproteinization and deproteinization, were separated on Sephadex G-200 gel permeation chromatography. The absorption curves of elution for the PSs were obtained by colorimetry, and the proteins were detected using Coomassic brilliant G-250. Eight REEs (La, Ce, Nd, Sm, Eu, Tb, Yb and Lu) in these PSs were determined by instrumental neutron activation analysis. The results obtained show that the REEs are bound firmly with the water-soluble PSs in the plant. A measurement demonstrates that the PSs bound with REEs are mainly of smaller molecular weight (10,000 to 20,000 Dalton)

  18. Transient intermediates are populated in the folding pathways of single-domain two-state folding protein L

    Science.gov (United States)

    Maity, Hiranmay; Reddy, Govardhan

    2018-04-01

    Small single-domain globular proteins, which are believed to be dominantly two-state folders, played an important role in elucidating various aspects of the protein folding mechanism. However, recent single molecule fluorescence resonance energy transfer experiments [H. Y. Aviram et al. J. Chem. Phys. 148, 123303 (2018)] on a single-domain two-state folding protein L showed evidence for the population of an intermediate state and it was suggested that in this state, a β-hairpin present near the C-terminal of the native protein state is unfolded. We performed molecular dynamics simulations using a coarse-grained self-organized-polymer model with side chains to study the folding pathways of protein L. In agreement with the experiments, an intermediate is populated in the simulation folding pathways where the C-terminal β-hairpin detaches from the rest of the protein structure. The lifetime of this intermediate structure increased with the decrease in temperature. In low temperature conditions, we also observed a second intermediate state, which is globular with a significant fraction of the native-like tertiary contacts satisfying the features of a dry molten globule.

  19. Chemical Compositions of Stars in the Globular Cluster NGC 3201: Tracers of Multi-Epoch Star Formation

    Science.gov (United States)

    Simmerer, Jennifer A.; Ivans, I. I.; Filler, D.

    2012-01-01

    The retrograde halo globular cluster NGC 3201 contains stars of substantially different iron abundance ([Fe/H]), a property that puts it at odds with the vast majority of the Galactic cluster system. Though its unusual orbit prompted speculation that NGC 3201 was the remnant of a captured object, much like the multi-metallicity globular cluster Omega Centauri, NGC 3201 is much less massive than Omega Centauri and all of the other halo globular clusters that have internal metallicity variations. We present the abundances of 21 elements in 24 red giant branch stars in NGC 3201 based on high-resolution (R 40,000), high signal-to-noise (S/N 70) spectra. We find that the detailed abundance pattern of NGC 3201 is unique amongst multi-metallicity halo clusters. Unlike M22, Omega Centauri, and NGC 1851, neither metal-poor nor metal-rich stars show any evidence of s-process enrichment (a product of the advanced evolution of low- and intermediate-mass stars). We find that while Na, O, and Al vary from star to star as is typical in globular clusters, there is no systematic difference between the abundance pattern in the metal-poor cluster stars and that of the metal-rich cluster stars. Furthermore, we find that the metallicity variations in NGC 3201 are independent of the well-known Na-O anticorrelation, which separates it from every other multi-metallicity cluster. In the context of a multi-episode star formation model, this implies that NGC 3201 began life with the [Fe/H] variations we measure now.

  20. Improved feed protein fractionation schemes for formulating rations with the cornell net carbohydrate and protein system.

    Science.gov (United States)

    Lanzas, C; Broderick, G A; Fox, D G

    2008-12-01

    Adequate predictions of rumen-degradable protein (RDP) and rumen-undegradable protein (RUP) supplies are necessary to optimize performance while minimizing losses of excess nitrogen (N). The objectives of this study were to evaluate the original Cornell Net Carbohydrate Protein System (CNCPS) protein fractionation scheme and to develop and evaluate alternatives designed to improve its adequacy in predicting RDP and RUP. The CNCPS version 5 fractionates CP into 5 fractions based on solubility in protein precipitant agents, buffers, and detergent solutions: A represents the soluble nonprotein N, B1 is the soluble true protein, B2 represents protein with intermediate rates of degradation, B3 is the CP insoluble in neutral detergent solution but soluble in acid detergent solution, and C is the unavailable N. Model predictions were evaluated with studies that measured N flow data at the omasum. The N fractionation scheme in version 5 of the CNCPS explained 78% of the variation in RDP with a root mean square prediction error (RMSPE) of 275 g/d, and 51% of the RUP variation with RMSPE of 248 g/d. Neutral detergent insoluble CP flows were overpredicted with a mean bias of 128 g/d (40% of the observed mean). The greatest improvements in the accuracy of RDP and RUP predictions were obtained with the following 2 alternative schemes. Alternative 1 used the inhibitory in vitro system to measure the fractional rate of degradation for the insoluble protein fraction in which A = nonprotein N, B1 = true soluble protein, B2 = insoluble protein, C = unavailable protein (RDP: R(2) = 0.84 and RMSPE = 167 g/d; RUP: R(2) = 0.61 and RMSPE = 209 g/d), whereas alternative 2 redefined A and B1 fractions as the non-amino-N and amino-N in the soluble fraction respectively (RDP: R(2) = 0.79 with RMSPE = 195 g/d and RUP: R(2) = 0.54 with RMSPE = 225 g/d). We concluded that implementing alternative 1 or 2 will improve the accuracy of predicting RDP and RUP within the CNCPS framework.

  1. Fluid-crystal coexistence for proteins and inorganic nanocolloids : Dependence on ionic strength

    NARCIS (Netherlands)

    Prinsen, P.; Odijk, T.

    2006-01-01

    We investigate theoretically the fluid-crystal coexistence of solutions of globular charged nanoparticles such as proteins and inorganic colloids. The thermodynamic properties of the fluid phase are computed via the optimized Baxter model P. Prinsen and T. Odijk [J. Chem. Phys. 121, 6525 (2004)].

  2. Sequential unfolding of the two-domain protein Pseudomonas stutzeri cytochrome c(4)

    DEFF Research Database (Denmark)

    Andersen, Niels Højmark; Jensen, Thomas Jon; Nørgaard, Allan

    2002-01-01

    F stutzeri cytochrome c. is a di-haem protein, composed of two globular domains each with His-Met coordinated haem. and a hydrogen bond network between the domains. The domain foldings are highly symmetric but with specific differences including structural differences of ligand coordination, and ...

  3. Intracellular transport of fat-soluble vitamins A and E.

    Science.gov (United States)

    Kono, Nozomu; Arai, Hiroyuki

    2015-01-01

    Vitamins are compounds that are essential for the normal growth, reproduction and functioning of the human body. Of the 13 known vitamins, vitamins A, D, E and K are lipophilic compounds and are therefore called fat-soluble vitamins. Because of their lipophilicity, fat-soluble vitamins are solubilized and transported by intracellular carrier proteins to exert their actions and to be metabolized properly. Vitamin A and its derivatives, collectively called retinoids, are solubilized by intracellular retinoid-binding proteins such as cellular retinol-binding protein (CRBP), cellular retinoic acid-binding protein (CRABP) and cellular retinal-binding protein (CRALBP). These proteins act as chaperones that regulate the metabolism, signaling and transport of retinoids. CRALBP-mediated intracellular retinoid transport is essential for vision in human. α-Tocopherol, the main form of vitamin E found in the body, is transported by α-tocopherol transfer protein (α-TTP) in hepatic cells. Defects of α-TTP cause vitamin E deficiency and neurological disorders in humans. Recently, it has been shown that the interaction of α-TTP with phosphoinositides plays a critical role in the intracellular transport of α-tocopherol and is associated with familial vitamin E deficiency. In this review, we summarize the mechanisms and biological significance of the intracellular transport of vitamins A and E. © 2014 The Authors. Traffic published by John Wiley & Sons Ltd.

  4. Medium Resolution Spectroscopy and Chemical Composition of Galactic Globular Clusters

    Directory of Open Access Journals (Sweden)

    Khamidullina D. A.

    2014-12-01

    Full Text Available We used integrated-light medium-resolution spectra of six Galactic globular clusters and model stellar atmospheres to carry out population synthesis and to derive chemical composition and age of the clusters. We used medium-resolution spectra of globular clusters published by Schiavon et al. (2005, as well as our long-slit observations with the 1.93 m telescope of the Haute Provence Observatory. The observed spectra were fitted to the theoretical ones interactively. As an initial approach, we used masses, radii and log g of stars in the clusters corresponding to the best fitting isochrones in the observed color-magnitude diagrams. The computed synthetic blanketed spectra of stars were summed according to the Chabrier mass function. To improve the determination of age and helium content, the shape and depth of the Balmer absorption lines was analysed. The abundances of Mg, Ca, C and several other elements were derived. A reasonable agreement with the literature data both in chemical composition and in age of the clusters is found. Our method might be useful for the development of stellar population models and for a better understanding of extragalactic star clusters.

  5. The Age of the Inner Halo Globular Cluster NGC 6652

    Science.gov (United States)

    Chaboyer, Brian; Sarajedini, Ata; Armandroff, Taft E.

    2000-01-01

    Hubble Space Telescope (HST) (V,I) photometry has been obtained for the inner halo globular cluster NGC 6652. The photometry reaches approximately 4 mag below the turn-off and includes a well populated horizontal branch (HB). This cluster is located close to the Galactic center at RGC approximately equal to 2.0 kpc with a reddening of E(V-I) = 0.15 +/- 0.02 and has a metallicity of [Fe/H] approximately equal to -0.85. Based upon DELTA V (sup SGB) (sub HB), NGC 6652 is 11.7 plus or minus 1.6 Gyr old. Using A HB precise differential ages for 47 Tuc (a thick disk globular), M107 and NGC 1851 (both halo clusters) were obtained. NGC 6652 appears to be the same age as 47 Tuc and NGC 1851 (within +/- 1.2 Gyr), while there is a slight suggestion that M107 is older than NGC 6652 by 2.3 +/- 1.5 Gyr. As this is a less than 2 sigma result, this issue needs to be investigated further before a definitive statement regarding the relative age of M107 and NGC 6652 may be made.

  6. Medium resolution spectroscopy and chemical composition of Galactic globular clusters

    Science.gov (United States)

    Khamidullina, D. A.; Sharina, M. E.; Shimansky, V. V.; Davoust, E.

    We used integrated-light medium-resolution spectra of six Galactic globular clusters and model stellar atmospheres to carry out population synthesis and to derive chemical composition and age of the clusters. We used medium-resolution spectra of globular clusters published by Schiavon et al. (2005), as well as our long-slit observations with the 1.93 m telescope of the Haute Provence Observatory. The observed spectra were fitted to the theoretical ones interactively. As an initial approach, we used masses, radii and log g of stars in the clusters corresponding to the best fitting isochrones in the observed color-magnitude diagrams. The computed synthetic blanketed spectra of stars were summed according to the Chabrier mass function. To improve the determination of age and helium content, the shape and depth of the Balmer absorption lines was analysed. The abundances of Mg, Ca, C and several other elements were derived. A reasonable agreement with the literature data both in chemical composition and in age of the clusters is found. Our method might be useful for the development of stellar population models and for a better understanding of extragalactic star clusters.

  7. Dynamical Friction in Multi-component Evolving Globular Clusters

    Science.gov (United States)

    Alessandrini, Emiliano; Lanzoni, Barbara; Miocchi, Paolo; Ciotti, Luca; Ferraro, Francesco R.

    2014-11-01

    We use the Chandrasekhar formalism and direct N-body simulations to study the effect of dynamical friction on a test object only slightly more massive than the field stars, orbiting a spherically symmetric background of particles with a mass spectrum. The main goal is to verify whether the dynamical friction time (t DF) develops a non-monotonic radial dependence that could explain the bimodality of the blue straggler radial distributions observed in globular clusters. In these systems, in fact, relaxation effects lead to a mass and velocity radial segregation of the different mass components, so that mass-spectrum effects on t DF are expected to be dependent on radius. We find that in spite of the presence of different masses, t DF is always a monotonic function of radius, at all evolutionary times and independently of the initial concentration of the simulated cluster. This is because the radial dependence of t DF is largely dominated by the total mass density profile of the background stars (which is monotonically decreasing with radius). Hence, a progressive temporal erosion of the blue straggler star (BSS) population at larger and larger distances from the cluster center remains the simplest and the most likely explanation of the shape of the observed BSS radial distributions, as suggested in previous works. We also confirm the theoretical expectation that approximating a multi-mass globular cluster as made of (averaged) equal-mass stars can lead to significant overestimations of t DF within the half-mass radius.

  8. Constraints from stellar models on mixing as a viable explanation of abundance anomalies in globular clusters

    International Nuclear Information System (INIS)

    Vandenberg, D.A.; Smith, G.H.

    1988-01-01

    Published observational data on changes in the surface abundances of evolving stars in globular clusters are compiled and compared with the predictions of theoretical evolutionary sequences (for stars of mass 0.8 solar mass and metallicity Z = 0.0001 or mass 0.9 solar mass and Z = 0.006) and of models incorporating enhanced envelope-interior mixing at various evolutionary phases. The results are presented in graphs and characterized in detail. It is found that mixing models of CN bimodality in globular-cluster stars can encounter difficulties when abundance anomalies appear early in the evolution of the star. 63 references

  9. Globular cluster neutron stars and the determination of the dense matter equation of state

    Science.gov (United States)

    Guillot, Sebastien

    2016-09-01

    Combining measurements of the mass and radius of multiple neutron stars (NSs) represents the most promising way to determine the equation of state of dense NS matter. NSs in quiescent low-mass x-ray binaries (qLMXB) located in globular clusters have placed useful constraints on the equation of state. The statistical approaches combining measurements from multiple NSs can be further improved by the addition of more NS observations. We propose here to obtain a high signal to noise spectrum of the qLMXB in M30, the only low-absorption globular cluster qLMXBs that does not have deep X-ray observations, and which requires Chandra unmatched angular resolution. The 300 ks proposed observation will permit measurement of the NS radius with 12-15% uncertainties.

  10. Extractable protein of radiation vulcanized natural rubber latex

    International Nuclear Information System (INIS)

    Soebianto, Y.S.; Ratnayake, U.M.; Makuuchi, Keizo; Yoshii, Fumio; Kume, Tamikazu

    2000-01-01

    Protein remained in the latex products are reported to cause serious allergy. A new method to reduce the protein level in the latex products by irradiation is reported. Water soluble protein (WSP) solution (10%) was added into radiation vulcanized NR latex (RVNRL) in three different processes. The amount of WSP was 3 phr. It was only added to RVNRL (standard), added to re-centrifuged RVNRL (pre-centrifugation), and added to RVNRL followed by centrifugation (post-centrifugation). The protein content was determined by enhanced BCA method, and identified by SDS-PAGE. Extractable protein (EP) from the rubber has been reduced up to the minimum protein detection by combining WSP addition and centrifugation. Short leaching time (20-30 min.) can be achieved after the combine treatment, and SDS-PAGE confirms the reduction of soluble protein in the serum phase, and disappearance of protein bands in the rubber extract. Protein-WSP interaction produces water soluble complex, and removed by centrifugation. The efficiency of protein removal by WSP depends on its molecular weight of WSP which relates to its water solubility. (author)

  11. Proteins at fluid interfaces: adsorption layers and thin liquid films.

    Science.gov (United States)

    Yampolskaya, Galina; Platikanov, Dimo

    2006-12-21

    A review in which many original published results of the authors as well as many other papers are discussed. The structure and some properties of the globular proteins are shortly presented, special accent being put on the alpha-chymotrypsin (alpha-ChT), lysozyme (LZ), human serum albumin (HSA), and bovine serum albumin (BSA) which have been used in the experiments with thin liquid films. The behaviour of protein adsorption layers (PAL) is extensively discussed. The dynamics of PAL formation, including the kinetics of adsorption as well as the time evolution of the surface tension of protein aqueous solutions, are considered. A considerable place is devoted to the surface tension and adsorption isotherms of the globular protein solutions, the simulation of PAL by interacting hard spheres, the experimental surface tension isotherms of the above mentioned proteins, and the interfacial tension isotherms for the protein aqueous solution/oil interface. The rheological properties of PAL at fluid interfaces are shortly reviewed. After a brief information about the experimental methods for investigation of protein thin liquid (foam or emulsion) films, the properties of the protein black foam films are extensively discussed: the conditions for their formation, the influence of the electrolytes and pH on the film type and stability, the thermodynamic properties of the black foam films, the contact angles film/bulk and their dynamic hysteresis. The next center of attention concerns some properties of the protein emulsion films: the conditions for formation of emulsion black films, the formation and development of a dimpling in microscopic, circular films. The protein-phospholipid mixed foam films are also briefly considered.

  12. Functional discrimination of membrane proteins using machine learning techniques

    Directory of Open Access Journals (Sweden)

    Yabuki Yukimitsu

    2008-03-01

    Full Text Available Abstract Background Discriminating membrane proteins based on their functions is an important task in genome annotation. In this work, we have analyzed the characteristic features of amino acid residues in membrane proteins that perform major functions, such as channels/pores, electrochemical potential-driven transporters and primary active transporters. Results We observed that the residues Asp, Asn and Tyr are dominant in channels/pores whereas the composition of hydrophobic residues, Phe, Gly, Ile, Leu and Val is high in electrochemical potential-driven transporters. The composition of all the amino acids in primary active transporters lies in between other two classes of proteins. We have utilized different machine learning algorithms, such as, Bayes rule, Logistic function, Neural network, Support vector machine, Decision tree etc. for discriminating these classes of proteins. We observed that most of the algorithms have discriminated them with similar accuracy. The neural network method discriminated the channels/pores, electrochemical potential-driven transporters and active transporters with the 5-fold cross validation accuracy of 64% in a data set of 1718 membrane proteins. The application of amino acid occurrence improved the overall accuracy to 68%. In addition, we have discriminated transporters from other α-helical and β-barrel membrane proteins with the accuracy of 85% using k-nearest neighbor method. The classification of transporters and all other proteins (globular and membrane showed the accuracy of 82%. Conclusion The performance of discrimination with amino acid occurrence is better than that with amino acid composition. We suggest that this method could be effectively used to discriminate transporters from all other globular and membrane proteins, and classify them into channels/pores, electrochemical and active transporters.

  13. Soluble Aβ aggregates can inhibit prion propagation.

    Science.gov (United States)

    Sarell, Claire J; Quarterman, Emma; Yip, Daniel C-M; Terry, Cassandra; Nicoll, Andrew J; Wadsworth, Jonathan D F; Farrow, Mark A; Walsh, Dominic M; Collinge, John

    2017-11-01

    Mammalian prions cause lethal neurodegenerative diseases such as Creutzfeldt-Jakob disease (CJD) and consist of multi-chain assemblies of misfolded cellular prion protein (PrP C ). Ligands that bind to PrP C can inhibit prion propagation and neurotoxicity. Extensive prior work established that certain soluble assemblies of the Alzheimer's disease (AD)-associated amyloid β-protein (Aβ) can tightly bind to PrP C , and that this interaction may be relevant to their toxicity in AD. Here, we investigated whether such soluble Aβ assemblies might, conversely, have an inhibitory effect on prion propagation. Using cellular models of prion infection and propagation and distinct Aβ preparations, we found that the form of Aβ assemblies which most avidly bound to PrP in vitro also inhibited prion infection and propagation. By contrast, forms of Aβ which exhibit little or no binding to PrP were unable to attenuate prion propagation. These data suggest that soluble aggregates of Aβ can compete with prions for binding to PrP C and emphasize the bidirectional nature of the interplay between Aβ and PrP C in Alzheimer's and prion diseases. Such inhibitory effects of Aβ on prion propagation may contribute to the apparent fall-off in the incidence of sporadic CJD at advanced age where cerebral Aβ deposition is common. © 2017 The Authors.

  14. Synthetic properties of models of globular clusters

    Energy Technology Data Exchange (ETDEWEB)

    Angeletti, L; Dolcetta, R; Giannone, P. (Rome Univ. (Italy). Osservatorio Astronomico)

    1980-05-01

    Synthetic and projected properties of models of globular clusters have been computed on the basis of stellar evolution and time changes of the dynamical cluster structure. Clusters with five and eight stellar groups (each group consisting of stars with the same mass) were studied. Mass loss from evolved stars was taken into account. Observational features were obtained at ages of 10-19 x 10/sup 9/ yr. The basic importance of the horizontal- and asymptotic-branch stars was pointed out. A comparison of the results with observed data of M3 is discussed with the purpose of obtaining general indications rather than a specific fit.

  15. Synthetic properties of models of globular clusters

    International Nuclear Information System (INIS)

    Angeletti, L.; Dolcetta, R.; Giannone, P.

    1980-01-01

    Synthetic and projected properties of models of globular clusters have been computed on the basis of stellar evolution and time changes of the dynamical cluster structure. Clusters with five and eight stellar groups (each group consisting of stars with the same mass) were studied. Mass loss from evolved stars was taken into account. Observational features were obtained at ages of 10-19 x 10 9 yr. The basic importance of the horizontal- and asymptotic-branch stars was pointed out. A comparison of the results with observed data of M3 is discussed with the purpose of obtaining general indications rather than a specific fit. (orig.)

  16. A systematic evaluation of solubility enhancing excipients to enable the generation of permeability data for poorly soluble compounds in Caco-2 model.

    Science.gov (United States)

    Shah, Devang; Paruchury, Sundeep; Matta, Muralikrishna; Chowan, Gajendra; Subramanian, Murali; Saxena, Ajay; Soars, Matthew G; Herbst, John; Haskell, Roy; Marathe, Punit; Mandlekar, Sandhya

    2014-01-01

    The study presented here identified and utilized a panel of solubility enhancing excipients to enable the generation of flux data in the Human colon carcinoma (Caco-2) system for compounds with poor solubility. Solubility enhancing excipients Dimethyl acetamide (DMA) 1 % v/v, polyethylene glycol (PEG) 400 1% v/v, povidone 1% w/v, poloxamer 188 2.5% w/v and bovine serum albumin (BSA) 4% w/v did not compromise Caco-2 monolayer integrity as assessed by trans-epithelial resistance measurement (TEER) and Lucifer yellow (LY) permeation. Further, these excipients did not affect P-glycoprotein (P-gp) mediated bidirectional transport of digoxin, permeabilities of high (propranolol) or low permeability (atenolol) compounds, and were found to be inert to Breast cancer resistant protein (BCRP) mediated transport of cladribine. This approach was validated further using poorly soluble tool compounds, atazanavir (poloxamer 188 2.5% w/v) and cyclosporine A (BSA 4% w/v) and also applied to new chemical entity (NCE) BMS-A in BSA 4% w/v, for which Caco-2 data could not be generated using the traditional methodology due to poor solubility (solubility of atazanavir by >8 fold whereas BSA 4% w/v increased the solubility of cyclosporine A and BMS-A by >2-4 fold thereby enabling permeability as well as efflux liability estimation in the Caco-2 model with reasonable recovery values. To conclude, addition of excipients such as poloxamer 188 2.5% w/v and BSA 4% w/v to HBSS leads to a significant improvement in the solubility of the poorly soluble compounds resulting in enhanced recoveries without modulating transporter-mediated efflux, expanding the applicability of Caco-2 assays to poorly soluble compounds.

  17. Enhancing the productivity of soluble green fluorescent protein ...

    African Journals Online (AJOL)

    Protein sequences might have been evolved against different environmental pressures, which results in non-optimum properties in their stability, activity and folding efficiency. Directed evolution and consensus-based engineering of proteins are the protein engineering principles for the re-evolution of such natural proteins ...

  18. Study of the hydration of globular proteins by broad NMR lines method

    Energy Technology Data Exchange (ETDEWEB)

    Blicharska, B [Uniwersytet Jagiellonski, Krakow (Poland). Instytut Fizyki

    1973-01-01

    Spectra of proteins and polypeptides obtained by means of a NMR broad line spectrometer consist of broad and thin lines. These broad and thin lines are attributed to proteins and to water absorbed on the surfaces of proteins respectively. The behaviour of the thin line in the spectra of lyophilizated albumin of the egg white has been studied in the temperature range from -42 to 20/sup 0/C. The amount of water has been found by the simple method of weighing and has been equal about 7% of the total weight. It has been found that the water absorbed on the surface of the lyophilizated proteins gives a thinner line in comparison to the water absorbed on molecules of proteins in water solutions and that the correlation time is about 10/sup 3/ times greater.

  19. Brain asymmetry in the white matter making and globularity

    Directory of Open Access Journals (Sweden)

    Constantina eTheofanopoulou

    2015-09-01

    Full Text Available Recent studies from the field of language genetics and evolutionary anthropology have put forward the hypothesis that the emergence of our species-specific brain is to be understood not in terms of size, but in light of developmental changes that gave rise to a more globular braincase configuration after the split from Neanderthals-Denisovans. On the grounds that (i white matter myelination is delayed relative to other brain structures and in humans is protracted compared with other primates and (ii neural connectivity is linked genetically to our brain/skull morphology and language-ready brain, I take it that one significant evolutionary change in Homo sapiens’ lineage is the interhemispheric connectivity mediated by the Corpus Callosum. The size, myelination and fiber caliber of the Corpus Callosum presents an anterior-to-posterior increase, in a way that inter-hemispheric connectivity is more prominent in the sensory motor areas, whereas high- order areas are more intra-hemispherically connected. Building on evidence from language-processing studies that account for this asymmetry (‘lateralization’ in terms of brain rhythms, I present an evo-devo hypothesis according to which the myelination of the Corpus Callosum, Brain Asymmetry and Globularity are conjectured to make up the angles of a co-evolutionary triangle that gave rise to our language-ready brain.

  20. Study of the spray to globular transition in gas metal arc welding: a spectroscopic investigation

    International Nuclear Information System (INIS)

    Valensi, F; Pellerin, S; Castillon, Q; Zielinska, S; Boutaghane, A; Dzierzega, K; Pellerin, N; Briand, F

    2013-01-01

    The gas metal arc welding (GMAW) process is strongly influenced by the composition of the shielding gas. In particular, addition of CO 2 increases the threshold current for the transition from unstable globular to more stable spray transfer mode. We report on the diagnostics—using optical emission spectroscopy—of a GMAW plasma in pure argon and in mixtures of argon, CO 2 and N 2 while operated in spray and globular transfer modes. The spatially resolved plasma parameters are obtained by applying the Abel transformation to laterally integrated emission data. The Stark widths of some iron lines are used to determine both electron density and temperature, and line intensities yield relative contents of neutral and ionized iron to argon. Our experimental results indicate a temperature drop on the arc axis in the case of spray arc transfer. This drop reduces with addition of N 2 and disappears in globular transfer mode when CO 2 is added. Despite the temperature increase, the electron density decreases with CO 2 concentration. The highest concentration of iron is observed in the plasma column upper part (close to the anode) and for GMAW with CO 2 . Our results are compared with recently published works where the effect of non-homogeneous metal vapour concentration has been taken into account. (paper)

  1. Improving recombinant protein solubility in Escherichia coli ...

    African Journals Online (AJOL)

    user

    2010-11-22

    Nov 22, 2010 ... protein (Baneyx and Mujacic, 2004). Although ... both time-consuming and expensive (Tsumoto et al.,. 2003). Thus ... proteins (Schlieker et al., 2002). ..... Alibolandi M, Mirzahoseini H, Abedi Khalil Abad M, Azami Movahed M.

  2. Characterization of a soluble phosphatidic acid phosphatase in bitter melon (Momordica charantia).

    Science.gov (United States)

    Cao, Heping; Sethumadhavan, Kandan; Grimm, Casey C; Ullah, Abul H J

    2014-01-01

    Momordica charantia is often called bitter melon, bitter gourd or bitter squash because its fruit has a bitter taste. The fruit has been widely used as vegetable and herbal medicine. Alpha-eleostearic acid is the major fatty acid in the seeds, but little is known about its biosynthesis. As an initial step towards understanding the biochemical mechanism of fatty acid accumulation in bitter melon seeds, this study focused on a soluble phosphatidic acid phosphatase (PAP, 3-sn-phosphatidate phosphohydrolase, EC 3.1.3.4) that hydrolyzes the phosphomonoester bond in phosphatidate yielding diacylglycerol and P(i). PAPs are typically categorized into two subfamilies: Mg(2+)-dependent soluble PAP and Mg(2+)-independent membrane-associated PAP. We report here the partial purification and characterization of an Mg(2+)-independent PAP activity from developing cotyledons of bitter melon. PAP protein was partially purified by successive centrifugation and UNOsphere Q and S columns from the soluble extract. PAP activity was optimized at pH 6.5 and 53-60 °C and unaffected by up to 0.3 mM MgCl2. The K(m) and Vmax values for dioleoyl-phosphatidic acid were 595.4 µM and 104.9 ηkat/mg of protein, respectively. PAP activity was inhibited by NaF, Na(3)VO(4), Triton X-100, FeSO4 and CuSO4, but stimulated by MnSO4, ZnSO4 and Co(NO3)2. In-gel activity assay and mass spectrometry showed that PAP activity was copurified with a number of other proteins. This study suggests that PAP protein is probably associated with other proteins in bitter melon seeds and that a new class of PAP exists as a soluble and Mg(2+)-independent enzyme in plants.

  3. SAXS analysis of a soluble cytosolic NgBR construct including extracellular and transmembrane domains.

    Directory of Open Access Journals (Sweden)

    Joshua Holcomb

    Full Text Available The Nogo-B receptor (NgBR is involved in oncogenic Ras signaling through directly binding to farnesylated Ras. It recruits farnesylated Ras to the non-lipid-raft membrane for interaction with downstream effectors. However, the cytosolic domain of NgBR itself is only partially folded. The lack of several conserved secondary structural elements makes this domain unlikely to form a complete farnesyl binding pocket. We find that inclusion of the extracellular and transmembrane domains that contain additional conserved residues to the cytosolic region results in a well folded protein with a similar size and shape to the E.coli cis-isoprenyl transferase (UPPs. Small Angle X-ray Scattering (SAXS analysis reveals the radius of gyration (Rg of our NgBR construct to be 18.2 Å with a maximum particle dimension (Dmax of 61.0 Å. Ab initio shape modeling returns a globular molecular envelope with an estimated molecular weight of 23.0 kD closely correlated with the calculated molecular weight. Both Kratky plot and pair distribution function of NgBR scattering reveal a bell shaped peak which is characteristic of a single globularly folded protein. In addition, circular dichroism (CD analysis reveals that our construct has the secondary structure contents similar to the UPPs. However, this result does not agree with the currently accepted topological orientation of NgBR which might partition this construct into three separate domains. This discrepancy suggests another possible NgBR topology and lends insight into a potential molecular basis of how NgBR facilitates farnesylated Ras recruitment.

  4. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors required during Trypanosoma cruzi parasitophorous vacuole development.

    Science.gov (United States)

    Cueto, Juan Agustín; Vanrell, María Cristina; Salassa, Betiana Nebaí; Nola, Sébastien; Galli, Thierry; Colombo, María Isabel; Romano, Patricia Silvia

    2017-06-01

    Trypanosoma cruzi, the etiologic agent of Chagas disease, is an obligate intracellular parasite that exploits different host vesicular pathways to invade the target cells. Vesicular and target soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are key proteins of the intracellular membrane fusion machinery. During the early times of T. cruzi infection, several vesicles are attracted to the parasite contact sites in the plasma membrane. Fusion of these vesicles promotes the formation of the parasitic vacuole and parasite entry. In this work, we study the requirement and the nature of SNAREs involved in the fusion events that take place during T. cruzi infection. Our results show that inhibition of N-ethylmaleimide-sensitive factor protein, a protein required for SNARE complex disassembly, impairs T. cruzi infection. Both TI-VAMP/VAMP7 and cellubrevin/VAMP3, two v-SNAREs of the endocytic and exocytic pathways, are specifically recruited to the parasitophorous vacuole membrane in a synchronized manner but, although VAMP3 is acquired earlier than VAMP7, impairment of VAMP3 by tetanus neurotoxin fails to reduce T. cruzi infection. In contrast, reduction of VAMP7 activity by expression of VAMP7's longin domain, depletion by small interfering RNA or knockout, significantly decreases T. cruzi infection susceptibility as a result of a minor acquisition of lysosomal components to the parasitic vacuole. In addition, overexpression of the VAMP7 partner Vti1b increases the infection, whereas expression of a KIF5 kinesin mutant reduces VAMP7 recruitment to vacuole and, concomitantly, T. cruzi infection. Altogether, these data support a key role of TI-VAMP/VAMP7 in the fusion events that culminate in the T. cruzi parasitophorous vacuole development. © 2016 John Wiley & Sons Ltd.

  5. Generation of Soluble Advanced Glycation End Products Receptor (sRAGE)-Binding Ligands during Extensive Heat Treatment of Whey Protein/Lactose Mixtures Is Dependent on Glycation and Aggregation

    NARCIS (Netherlands)

    Liu, Fahui; Teodorowicz, Gosia; Wichers, Harry J.; Boekel, van Tiny; Hettinga, Kasper A.

    2016-01-01

    Heating of protein- and sugar-containing materials is considered the primary factor affecting the formation of advanced glycation end products (AGEs). This study aimed to investigate the influence of heating conditions, digestion, and aggregation on the binding capacity of AGEs to the soluble AGE

  6. The properties of the disk system of globular clusters

    Science.gov (United States)

    Armandroff, Taft E.

    1989-01-01

    A large refined data sample is used to study the properties and origin of the disk system of globular clusters. A scale height for the disk cluster system of 800-1500 pc is found which is consistent with scale-height determinations for samples of field stars identified with the Galactic thick disk. A rotational velocity of 193 + or - 29 km/s and a line-of-sight velocity dispersion of 59 + or - 14 km/s have been found for the metal-rich clusters.

  7. Photoelectric UBVRI sequences in the Galactic globular clusters NGC 6752 and NGC 6864

    International Nuclear Information System (INIS)

    Alvarado, F.; Wenderoth, E.; Alcaino, G.; Liller, W.

    1990-01-01

    UBVRI photoelectric sequences for the Galactic globular clusters NGC 6752 and NGC 6864 are presented. Both of them include fields suitable for CCD exposures. From five UBV sequences in NGC 6572, only five stars are in common with the previous works. 15 refs

  8. Simplified Method for Rapid Purification of Soluble Histones

    Directory of Open Access Journals (Sweden)

    Nives Ivić

    2016-06-01

    Full Text Available Functional and structural studies of histone-chaperone complexes, nucleosome modifications, their interactions with remodelers and regulatory proteins rely on obtaining recombinant histones from bacteria. In the present study, we show that co-expression of Xenopus laevis histone pairs leads to production of soluble H2AH2B heterodimer and (H3H42 heterotetramer. The soluble histone complexes are purified by simple chromatographic techniques. Obtained H2AH2B dimer and H3H4 tetramer are proficient in histone chaperone binding and histone octamer and nucleosome formation. Our optimized protocol enables rapid purification of multiple soluble histone variants with a remarkable high yield and simplifies histone octamer preparation. We expect that this simple approach will contribute to the histone chaperone and chromatin research. This work is licensed under a Creative Commons Attribution 4.0 International License.

  9. La galaxia NGC 6876 y su sistema de cúmulos globulares

    Science.gov (United States)

    Ennis, A. I.; Bassino, L. P.; Caso, J. P.

    2017-10-01

    We present preliminary results of the deep photometric study of the elliptical galaxy NGC6876, located at the center of the Pavo group, and its globular cluster system. We use images obtained with the GMOS camera mounted on the Gemini South telescope, in the and bands, with the purpose of disentangling the evolutionary history of the galaxy on the basis of their characteristics.

  10. Synthetic horizontal branch models for globular clusters - the luminosity of the horizontal branch and the Oosterhoff effect

    International Nuclear Information System (INIS)

    Lee, Y.W.; Demarque, P.; Zinn, R.

    1987-01-01

    The variation of horizontal-branch (HB) luminosities with metal abundances is analyzed on the basis of HB models synthesized from theoretical HB evolutionary tracks. The focus is on the Oosterhoff effect, as related to period shifts in globular-cluster RR Lyr variables. The construction of the models and the Oosterhoff period groups is explained in detail, and the implications for globular-cluster ages are considered. The ratio of Delta M(bol) (RR) to Delta Fe/H for the HB is calculated as 0.24, slightly steeper than that found by Sandage (1981 and 1982). 35 references

  11. Comparing Chemical Compositions of Dwarf Elliptical Galaxies and Globular Clusters

    Science.gov (United States)

    Chu, Jason; Sparkman, Lea; Toloba, Elisa; Guhathakurta, Puragra

    2015-01-01

    Because of their abundance in cluster environments and fragility due to their low mass, dwarf elliptical galaxies (dEs) are excellent specimens for studying the physical processes that occur inside galaxy clusters. These studies can be used to expand our understanding of the process of galaxy (specifically dE) formation and the role of dark matter in the Universe. To move closer to better understanding these topics, we present a study of the relationship between dEs and globular clusters (GCs) by using the largest sample of dEs and GC satellites to date. We focus on comparing the ages and chemical compositions of dE nuclei with those of satellite GCs by analyzing absorption lines in their spectra. To better view the spectral features of these relatively dim objects, we employ a spectral co-addition process, where we add the fluxes of several objects to produce a single spectrum with high signal-to-noise ratio. Our finding that dE nuclei are younger and more metal rich than globular clusters establishes important benchmarks that future dE formation theories will consider. We also establish a means to identify GCs whose parent galaxies are uncertain, which allows us to make comparisons between this GC group and the satellite GCs.

  12. Enzymeaticial analysis and soluble proteins assays on radioprotective effects of cordyceps militaris

    Energy Technology Data Exchange (ETDEWEB)

    Yoo, Beong Gyu [Wonkwang Health Science College, Iksan (Korea, Republic of); Park, Joon Chul [Ansan 1 College, Ansan (Korea, Republic of)

    2001-06-01

    Effect of single pre-administration of Cordyceps militaries (Cm) extract on the survival ratio, body weight and organ weight changes and blood cell counts after whole-body {gamma}-irradiation were investigated. The single pre-administration of Cm extract at 24 hrs before {gamma}-irradiation increased the 40-day survival ration of irradiated mice from 60.1% to 71/4%. The administration of Cm extract completely prevented weight reductions of spleen and thymus produced by {gamma}-irradiation (P<0.01, P<0.05). Similar but somewhat less radioprotective effect was also found in the testis of the Cm treated mice. The administration of Cm extract retarded the reduction of both leukocyte and lymphocyte counts occured during the first 7 days and accelerated the recovery of the counts thereafter. The extract also accelerated the recovery of the erythrocyte counts occurred after the day 21th. SDS-polyacrylamide gel electrophoresis of the soluble proteins extracted from various organs did not reveal differences to any extent in all groups except in the levers of the irradiated and extract treated groups, in which some proteins were missing or less present. Also, the result of general intra and extra mycelial enzyme assays with Cm, extramycelial enzyme activity was relatively higher than the intramycelial enzyme. Cm appeared to indicate that {alpha}-amylase was the highest among the enzymes and gluosidase and chitinase were followed. Since the spleen, thymus and testis have been well known as radiosensitive organs, the protective action of Cm extract on irradiated mice may be responsible for its enhancing recovery of these organs. Although the exact mechanism in protective effect of Cm extract on irradiated mice is not clear yet, the present study is the first report regarding the Cm which was tested and found to be a potential radioprotective agent.

  13. Enzymeaticial analysis and soluble proteins assays on radioprotective effects of cordyceps militaris

    International Nuclear Information System (INIS)

    Yoo, Beong Gyu; Park, Joon Chul

    2001-01-01

    Effect of single pre-administration of Cordyceps militaries (Cm) extract on the survival ratio, body weight and organ weight changes and blood cell counts after whole-body γ-irradiation were investigated. The single pre-administration of Cm extract at 24 hrs before γ-irradiation increased the 40-day survival ration of irradiated mice from 60.1% to 71/4%. The administration of Cm extract completely prevented weight reductions of spleen and thymus produced by γ-irradiation (P<0.01, P<0.05). Similar but somewhat less radioprotective effect was also found in the testis of the Cm treated mice. The administration of Cm extract retarded the reduction of both leukocyte and lymphocyte counts occured during the first 7 days and accelerated the recovery of the counts thereafter. The extract also accelerated the recovery of the erythrocyte counts occurred after the day 21th. SDS-polyacrylamide gel electrophoresis of the soluble proteins extracted from various organs did not reveal differences to any extent in all groups except in the levers of the irradiated and extract treated groups, in which some proteins were missing or less present. Also, the result of general intra and extra mycelial enzyme assays with Cm, extramycelial enzyme activity was relatively higher than the intramycelial enzyme. Cm appeared to indicate that α-amylase was the highest among the enzymes and gluosidase and chitinase were followed. Since the spleen, thymus and testis have been well known as radiosensitive organs, the protective action of Cm extract on irradiated mice may be responsible for its enhancing recovery of these organs. Although the exact mechanism in protective effect of Cm extract on irradiated mice is not clear yet, the present study is the first report regarding the Cm which was tested and found to be a potential radioprotective agent

  14. Extragalactic globular clusters. I. The metallicity calibration

    International Nuclear Information System (INIS)

    Brodie, J.P.; Huchra, J.P.

    1990-01-01

    The ability of absorption-line strength indices, measured from integrated globular cluster spectra, to predict mean cluster metallicity is explored. Statistical criteria, are used to identify the six best indices out of about 20 measured in a large sample of Galactic and M31 cluster spectra. Linear relations between index and metallicity have been derived along with new calibrations of infrared colors (V - K, J - K, and CO) versus Fe/H. Estimates of metallicity from the six spectroscopic index-metallicity relations have been combined in three different ways to identify the most efficient estimator and the minimum bias estimator of Fe/H - the weighted mean. This provides an estimate of Fe/H accurate to about 15 percent. 37 refs

  15. Multiple populations along the asymptotic giant branch of the globular cluster M 4

    NARCIS (Netherlands)

    Lardo, C.; Salaris, M.; Savino, A.; Donati, P.; Stetson, P. B.; Cassisi, S.

    2016-01-01

    Nearly all Galactic globular clusters host stars that display characteristic abundance anti-correlations, like the O-rich/Na-poor pattern typical of field halo stars, together with O-poor/Na-rich additional components. A recent spectroscopic investigation questioned the presence of O-poor/Na-rich

  16. Dietary fiber content influences soluble carbohydrate levels in ruminal fluids.

    Science.gov (United States)

    Pinder, R S; Patterson, J A; O'Bryan, C A; Crandall, P G; Ricke, S C

    2012-01-01

    The soluble carbohydrate concentration of ruminal fluid, as affected by dietary forage content (DFC) and/or ruminally undegradable intake protein content (UIPC), was determined. Four ruminally cannulated steers, in a 4 × 4 Latin square design, were offered diets containing high (75 % of DM) or low (25 % of DM) DFC and high (6 % of DM) or low (5 % of DM) UIPC, in a 2 × 2 factorial arrangement. Zinc-treated SBM was the primary UIP source. Soluble hexose concentration (145.1 μM) in ruminal fluid (RF) of steers fed low DFC diets exhibited a higher trend (P = 0.08) than that (124.5 μM) of steers fed high DFC diets. UIPC did not modulate (P = 0.54) ruminal soluble hexose concentrations. Regardless of diet, soluble hexose concentration declined immediately after feeding and did not rise until 3 h after feeding (P ruminal fluid could not be determined. However, unsubstituted xylose and arabinose were excluded. These data indicate that: (i) soluble carbohydrate concentrations remain in ruminal fluid during digestion and fermentation; (ii) slight diurnal changes began after feeding; (iii) DFC influences the soluble carbohydrate concentration in RF; and (iv) UIPC of these diets does not affect the soluble carbohydrate concentration of RF.

  17. Are tyrosine residues involved in the photoconversion of the water-soluble chlorophyll-binding protein of Chenopodium album?

    Science.gov (United States)

    Takahashi, S; Seki, Y; Uchida, A; Nakayama, K; Satoh, H

    2015-05-01

    Non-photosynthetic and hydrophilic chlorophyll (Chl) proteins, called water-soluble Chl-binding proteins (WSCPs), are distributed in various species of Chenopodiaceae, Amaranthaceae, Polygonaceae and Brassicaceae. Based on their photoconvertibility, WSCPs are categorised into two classes: Class I (photoconvertible) and Class II (non-photoconvertible). Chenopodium album WSCP (CaWSCP; Class I) is able to convert the chlorin skeleton of Chl a into a bacteriochlorin-like skeleton under light in the presence of molecular oxygen. Potassium iodide (KI) is a strong inhibitor of the photoconversion. Because KI attacks tyrosine residues in proteins, tyrosine residues in CaWSCP are considered to be important amino acid residues for the photoconversion. Recently, we identified the gene encoding CaWSCP and found that the mature region of CaWSCP contained four tyrosine residues: Tyr13, Tyr14, Tyr87 and Tyr134. To gain insight into the effect of the tyrosine residues on the photoconversion, we constructed 15 mutant proteins (Y13A, Y14A, Y87A, Y134A, Y13-14A, Y13-87A, Y13-134A, Y14-87A, Y14-134A, Y87-134A, Y13-14-87A, Y13-14-134A, Y13-87-134A, Y14-87-134A and Y13-14-87-134A) using site-directed mutagenesis. Amazingly, all the mutant proteins retained not only chlorophyll-binding activity, but also photoconvertibility. Furthermore, we found that KI strongly inhibited the photoconversion of Y13-14-87-134A. These findings indicated that the four tyrosine residues are not essential for the photoconversion. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

  18. Analysis of long-range correlation in sequences data of proteins

    Directory of Open Access Journals (Sweden)

    ADRIANA ISVORAN

    2007-04-01

    Full Text Available The results presented here suggest the existence of correlations in the sequence data of proteins. 32 proteins, both globular and fibrous, both monomeric and polymeric, were analyzed. The primary structures of these proteins were treated as time series. Three spatial series of data for each sequence of a protein were generated from numerical correspondences between each amino acid and a physical property associated with it, i.e., its electric charge, its polar character and its dipole moment. For each series, the spectral coefficient, the scaling exponent and the Hurst coefficient were determined. The values obtained for these coefficients revealed non-randomness in the series of data.

  19. A Wide-Field Photometric Survey for Extratidal Tails Around Five Metal-Poor Globular Clusters in the Galactic Halo

    Science.gov (United States)

    Chun, Sang-Hyun; Kim, Jae-Woo; Sohn, Sangmo T.; Park, Jang-Hyun; Han, Wonyong; Kim, Ho-Il; Lee, Young-Wook; Lee, Myung Gyoon; Lee, Sang-Gak; Sohn, Young-Jong

    2010-02-01

    Wide-field deep g'r'i' images obtained with the Megacam of the Canada-France-Hawaii Telescope are used to investigate the spatial configuration of stars around five metal-poor globular clusters M15, M30, M53, NGC 5053, and NGC 5466, in a field-of-view ~3°. Applying a mask filtering algorithm to the color-magnitude diagrams of the observed stars, we sorted cluster's member star candidates that are used to examine the characteristics of the spatial stellar distribution surrounding the target clusters. The smoothed surface density maps and the overlaid isodensity contours indicate that all of the five metal-poor globular clusters exhibit strong evidence of extratidal overdensity features over their tidal radii, in the form of extended tidal tails around the clusters. The orientations of the observed extratidal features show signatures of tidal tails tracing the clusters' orbits, inferred from their proper motions, and effects of dynamical interactions with the Galaxy. Our findings include detections of a tidal bridge-like feature and an envelope structure around the pair of globular clusters M53 and NGC 5053. The observed radial surface density profiles of target clusters have a deviation from theoretical King models, for which the profiles show a break at 0.5-0.7rt , extending the overdensity features out to 1.5-2rt . Both radial surface density profiles for different angular sections and azimuthal number density profiles confirm the overdensity features of tidal tails around the five metal-poor globular clusters. Our results add further observational evidence that the observed metal-poor halo globular clusters originate from an accreted satellite system, indicative of the merging scenario of the formation of the Galactic halo. Based on observations carried out at the Canada-France-Hawaii Telescope, operated by the National Research Council of Canada, the Centre National de la Recherche Scientifique de France, and the University of Hawaii. This is part of the

  20. Ages of Globular Clusters from HIPPARCOS Parallaxes of Local Subdwarfs

    Science.gov (United States)

    Gratton, Raffaele G.; Fusi Pecci, Flavio; Carretta, Eugenio; Clementini, Gisella; Corsi, Carlo E.; Lattanzi, Mario

    1997-12-01

    We report here initial but strongly conclusive results for absolute ages of Galactic globular clusters (GGCs). This study is based on high-precision trigonometric parallaxes from the HIPPARCOS satellite coupled with accurate metal abundances ([Fe/H], [O/Fe], and [α/Fe]) from high-resolution spectroscopy for a sample of about thirty subdwarfs. Systematic effects due to star selection (Lutz-Kelker corrections to parallaxes) and the possible presence of undetected binaries in the sample of bona fide single stars are examined, and appropriate corrections are estimated. They are found to be small for our sample. The new data allow us to reliably define the absolute location of the main sequence (MS) as a function of metallicity. These results are then used to derive distances and ages for a carefully selected sample of nine globular clusters having metallicities determined from high-dispersion spectra of individual giants according to a procedure totally consistent with that used for the field subdwarfs. Very precise and homogeneous reddening values have also been independently determined for these clusters. Random errors for our distance moduli are +/-0.08 mag, and systematic errors are likely of the same order of magnitude. These very accurate distances allow us to derive ages with internal errors of ~12% (+/-1.5 Gyr). The main results are: 1. HIPPARCOS parallaxes are smaller than corresponding ground-based measurements, leading, in turn, to longer distance moduli (~0.2 mag) and younger ages (~2.8 Gyr). 2. The distance to NGC 6752 derived from our MS fitting is consistent with that determined using the white dwarf cooling sequence. 3. The relation between the zero-age HB (ZAHB) absolute magnitude and metallicity for the nine program clusters is MV(ZAHB)=(0.22+/-0.09)([Fe/H]+1.5)+(0.49+/-0.04) . This relation is fairly consistent with some of the most recent theoretical models. Within quoted errors, the slope is in agreement with that given by the Baade-Wesselink (BW

  1. THE CENTRAL BLUE STRAGGLER POPULATION IN FOUR OUTER-HALO GLOBULAR CLUSTERS

    Energy Technology Data Exchange (ETDEWEB)

    Beccari, Giacomo; Luetzgendorf, Nora [European Southern Observatory, Karl-Schwarzschild-Strasse 2, 85748 Garching bei Muenchen (Germany); Olczak, Christoph [Astronomisches Rechen-Institut (ARI), Zentrum fuer Astronomie Universitaet Heidelberg, Moenchhofstrasse 1214, 69120 Heidelberg (Germany); Ferraro, Francesco R.; Lanzoni, Barbara [Dipartimento di Astronomia, Universita degli Studi di Bologna, via Ranzani 1, I-40127 Bologna (Italy); Carraro, Giovanni; Boffin, Henri M. J. [European Southern Observatory, Alonso de Cordova 3107, Santiago de Chile (Chile); Stetson, Peter B. [National Research Council of Canada, Herzberg Institute of Astrophysics, 5071 West Saanich Road, Victoria, BC V9E 2E7 (Canada); Sollima, Antonio [INAF-Osservatorio Astronomico di Padova, Vicolo dell' Osservatorio 5, 35122 Padova (Italy)

    2012-08-01

    Using Hubble Space Telescope/Wide Field Planetary Camera 2 data, we have performed a comparative study of the Blue Straggler Star (BSS) populations in the central regions of the globular clusters (GCs) AM 1, Eridanus, Palomar 3, and Palomar 4. Located at distances R{sub GC} > 50 kpc from the Galactic center, these are (together with Palomar 14 and NGC 2419) the most distant clusters in the halo. We determine their color-magnitude diagrams and centers of gravity. The four clusters turn out to have similar ages (10.5-11 Gyr), significantly smaller than those of the inner-halo globulars, and similar metallicities. By exploiting wide-field ground-based data, we build the most extended radial density profiles from resolved star counts ever published for these systems. These are well reproduced by isotropic King models of relatively low concentration. BSSs appear to be significantly more centrally segregated than red giants in all GCs, in agreement with the estimated core and half-mass relaxation times which are smaller than the cluster ages. Assuming that this is a signature of mass segregation, we conclude that AM 1 and Eridanus are slightly dynamically more evolved than Pal 3 and Pal 4.

  2. THE METALLICITY BIMODALITY OF GLOBULAR CLUSTER SYSTEMS: A TEST OF GALAXY ASSEMBLY AND OF THE EVOLUTION OF THE GALAXY MASS-METALLICITY RELATION

    International Nuclear Information System (INIS)

    Tonini, Chiara

    2013-01-01

    We build a theoretical model to study the origin of the globular cluster metallicity bimodality in the hierarchical galaxy assembly scenario. The model is based on empirical relations such as the galaxy mass-metallicity relation [O/H]-M star as a function of redshift, and on the observed galaxy stellar mass function up to redshift z ∼ 4. We make use of the theoretical merger rates as a function of mass and redshift from the Millennium simulation to build galaxy merger trees. We derive a new galaxy [Fe/H]-M star relation as a function of redshift, and by assuming that globular clusters share the metallicity of their original parent galaxy at the time of their formation, we populate the merger tree with globular clusters. We perform a series of Monte Carlo simulations of the galaxy hierarchical assembly, and study the properties of the final globular cluster population as a function of galaxy mass, assembly and star formation history, and under different assumptions for the evolution of the galaxy mass-metallicity relation. The main results and predictions of the model are the following. (1) The hierarchical clustering scenario naturally predicts a metallicity bimodality in the galaxy globular cluster population, where the metal-rich subpopulation is composed of globular clusters formed in the galaxy main progenitor around redshift z ∼ 2, and the metal-poor subpopulation is composed of clusters accreted from satellites, and formed at redshifts z ∼ 3-4. (2) The model reproduces the observed relations by Peng et al. for the metallicities of the metal-rich and metal-poor globular cluster subpopulations as a function of galaxy mass; the positions of the metal-poor and metal-rich peaks depend exclusively on the evolution of the galaxy mass-metallicity relation and the [O/Fe], both of which can be constrained by this method. In particular, we find that the galaxy [O/Fe] evolves linearly with redshift from a value of ∼0.5 at redshift z ∼ 4 to a value of ∼0.1 at

  3. The Case of the Missing Cyanogen-rich AGB Stars in Galactic Globular Clusters

    DEFF Research Database (Denmark)

    Campbell, S. W.; Yong, D.; Wylie-de Boer, E. C.

    2012-01-01

    The handful of available observations of AGB stars in Galactic Globular Clusters suggest that the GC AGB populations are dominated by cyanogen-weak stars (eg. Norris et al. 1981; Sneden et al. 2000). This contrasts strongly with the distributions on the RGB (and other) populations, which generall...

  4. Plutonium solubilities

    International Nuclear Information System (INIS)

    Puigdomnech, I.; Bruno, J.

    1991-02-01

    Thermochemical data has been selected for plutonium oxide, hydroxide, carbonate and phosphate equilibria. Equilibrium constants have been evaluated in the temperature range 0 to 300 degrees C at a pressure of 1 bar to T≤100 degrees C and at the steam saturated pressure at higher temperatures. Measured solubilities of plutonium that are reported in the literature for laboratory experiments have been collected. Solubility data on oxides, hydroxides, carbonates and phosphates have been selected. No solubility data were found at temperatures higher than 60 degrees C. The literature solubility data have been compared with plutonium solubilities calculated with the EQ3/6 geochemical modelling programs, using the selected thermodynamic data for plutonium. (authors)

  5. Extractable protein of radiation vulcanized natural rubber latex

    International Nuclear Information System (INIS)

    Soebianto, Y.S.; Upul, R.M.; Makuuchi, K.; Yoshii, F.; Kume, T.

    2000-01-01

    A new method to reduce the protein level in the latex products by irradiation is reported. Water soluble protein (WSP) solution (10%) was added into radiation vulcanized NR latex (RVNRL) as much as 3 phr in three different processes: added to RVNRL, added to re-centrifuged RVNRL, and added to RVNRL followed by centrifugation. The protein content was determined by enhanced BCA method, and identified by SDS-PAGE analysis. Addition of WSP followed by centrifugation reduces EP up to the minimum protein detection, and shortens the leaching time to 20-30 min. SDS-PAGE analysis confirms the reduction of soluble protein in the serum phase, and disappearance of protein bands in the rubber extract. Protein-WSP interaction produces water soluble complex, and removed by centrifugation. The molecular weight of WSP dictates the efficiency of protein removal. (author)

  6. Extractable protein of radiation vulcanized natural rubber latex

    Energy Technology Data Exchange (ETDEWEB)

    Soebianto, Y.S. [Center for Research and Development of Isotopes and Radiation Technology, BATAN, Jakarta (Indonesia); Upul, R.M. [Rubber Research Institute of Sri Lanka, Ratmalana (Sri Lanka); Makuuchi, K.; Yoshii, F.; Kume, T. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    A new method to reduce the protein level in the latex products by irradiation is reported. Water soluble protein (WSP) solution (10%) was added into radiation vulcanized NR latex (RVNRL) as much as 3 phr in three different processes: added to RVNRL, added to re-centrifuged RVNRL, and added to RVNRL followed by centrifugation. The protein content was determined by enhanced BCA method, and identified by SDS-PAGE analysis. Addition of WSP followed by centrifugation reduces EP up to the minimum protein detection, and shortens the leaching time to 20-30 min. SDS-PAGE analysis confirms the reduction of soluble protein in the serum phase, and disappearance of protein bands in the rubber extract. Protein-WSP interaction produces water soluble complex, and removed by centrifugation. The molecular weight of WSP dictates the efficiency of protein removal. (author)

  7. Globular domain of adiponectin: promising target molecule for detection of atherosclerotic lesions

    Science.gov (United States)

    Almer, Gunter; Saba-Lepek, Matthias; Haj-Yahya, Samih; Rohde, Eva; Strunk, Dirk; Fröhlich, Eleonore; Prassl, Ruth; Mangge, Harald

    2011-01-01

    Background: Adiponectin, an adipocyte-specific plasma protein, has been shown to accumulate in injured endothelial cells during development of atherosclerotic lesions. In this study, we investigated the potential of different adiponectin subfractions with special emphasis on globular adiponectin (gAd) to recognize and visualize atherosclerotic lesions. Methods: Recombinant mouse gAd and subfractions of full-length adiponectin (ie, trimeric, hexameric, and oligomeric forms) were fluorescence-labeled. Aortas of wild-type and apoprotein E-deficient mice fed a high cholesterol diet were dissected and incubated with the labeled biomarkers. Imaging was performed using confocal laser scanning microscopy. Results: Confocal laser scanning microscopic images showed that gAd binds more strongly to atherosclerotic plaques than full-length adiponectin subfractions. Further, we showed that gAd accumulates preferentially in endothelial cells and the fibrous cap area of plaques. Here we demonstrate for the first time that gAd recognizes atherosclerotic plaques on aortic sections of apoprotein E-deficient mice. Conclusion: These results suggest that gAd, in addition to its physiological properties, is also suitable as a target molecule for prospective diagnostic strategies in imaging atherosclerotic lesions. PMID:22022204

  8. THE DYNAMICAL EFFECTS OF WHITE DWARF BIRTH KICKS IN GLOBULAR STAR CLUSTERS

    International Nuclear Information System (INIS)

    Fregeau, John M.; Richer, Harvey B.; Rasio, Frederic A.; Hurley, Jarrod R.

    2009-01-01

    Recent observations of the white dwarf (WD) populations in the Galactic globular cluster NGC 6397 suggest that WDs receive a kick of a few km s -1 shortly before they are born. Using our Monte Carlo cluster evolution code, which includes accurate treatments of all relevant physical processes operating in globular clusters, we study the effects of the kicks on their host cluster and on the WD population itself. We find that in clusters whose velocity dispersion is comparable to the kick speed, WD kicks are a significant energy source for the cluster, prolonging the initial cluster core contraction phase significantly so that at late times the cluster core-to-half-mass radius ratio is a factor of up to ∼10 larger than in the no-kick case. WD kicks thus represent a possible resolution of the large discrepancy between observed and theoretically predicted values of this key structural parameter. Our modeling also reproduces the observed trend for younger WDs to be more extended in their radial distribution in the cluster than older WDs.

  9. A Fossil Bulge Globular Cluster Revealed by very Large Telescope Multi-conjugate Adaptive Optics

    Czech Academy of Sciences Publication Activity Database

    Ortolani, S.; Barbuy, B.; Momany, Y.; Saviane, I.; Bica, E.; Jílková, L.; Salerno, G.M.; Jungwiert, Bruno

    2011-01-01

    Roč. 737, č. 1 (2011), 31/1-31/9 ISSN 0004-637X Institutional research plan: CEZ:AV0Z10030501 Keywords : galaxy * globular clusters Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics Impact factor: 6.024, year: 2011

  10. No Evidence for Multiple Stellar Populations in the Low-mass Galactic Globular Cluster E 3

    Science.gov (United States)

    Salinas, Ricardo; Strader, Jay

    2015-08-01

    Multiple stellar populations are a widespread phenomenon among Galactic globular clusters. Even though the origin of the enriched material from which new generations of stars are produced remains unclear, it is likely that self-enrichment will be feasible only in clusters massive enough to retain this enriched material. We searched for multiple populations in the low mass (M˜ 1.4× {10}4 {M}⊙ ) globular cluster E3, analyzing SOAR/Goodman multi-object spectroscopy centered on the blue cyanogen (CN) absorption features of 23 red giant branch stars. We find that the CN abundance does not present the typical bimodal behavior seen in clusters hosting multistellar populations, but rather a unimodal distribution that indicates the presence of a genuine single stellar population, or a level of enrichment much lower than in clusters that show evidence for two populations from high-resolution spectroscopy. E3 would be the first bona fide Galactic old globular cluster where no sign of self-enrichment is found. Based on observations obtained at the Southern Astrophysical Research (SOAR) Telescope, which is a joint project of the Ministério da Ciência, Tecnologia, e Inovação (MCTI) da República Federativa do Brasil, the US National Optical Astronomy Observatory (NOAO), the University of North Carolina at Chapel Hill (UNC), and Michigan State University (MSU).

  11. Nutritive value of wet distillers' solubles for pigs

    Directory of Open Access Journals (Sweden)

    Jarmo Valaja

    1995-01-01

    Full Text Available Digestibility and nitrogen (N metabolism were studied to evaluate the nutritive value of wet barley distillers’ solubles (DSB from an integrated starch-ethanol process for pigs. Eight castrated male pigs (live weight 72-103 kg were used in a 8 x 3 cyclic change-over design, where the diets were arranged factorially 2x2. The corresponding factors were the protein source (DSB or soya bean meal (SBM and the protein level (131 or 162 g crude protein (CP/kg dry matter (DM. Faeces and urine were collected in total. The four diets comprised barley, barley starch, minerals and vitamins with either DSB or SBM as the main source of protein. The digestibility of CP(p

  12. Interactions of glycine betaine with proteins: insights from volume and compressibility measurements.

    Science.gov (United States)

    Shek, Yuen Lai; Chalikian, Tigran V

    2013-01-29

    We report the first application of volume and compressibility measurements to characterization of interactions between cosolvents (osmolytes) and globular proteins. Specifically, we measure the partial molar volumes and adiabatic compressibilities of cytochrome c, ribonuclease A, lysozyme, and ovalbumin in aqueous solutions of the stabilizing osmolyte glycine betaine (GB) at concentrations between 0 and 4 M. The fact that globular proteins do not undergo any conformational transitions in the presence of GB provides an opportunity to study the interactions of GB with proteins in their native states within the entire range of experimentally accessible GB concentrations. We analyze our resulting volumetric data within the framework of a statistical thermodynamic model in which each instance of GB interaction with a protein is viewed as a binding reaction that is accompanied by release of four water molecules. From this analysis, we calculate the association constants, k, as well as changes in volume, ΔV(0), and adiabatic compressibility, ΔK(S0), accompanying each GB-protein association event in an ideal solution. By comparing these parameters with similar characteristics determined for low-molecular weight analogues of proteins, we conclude that there are no significant cooperative effects involved in interactions of GB with any of the proteins studied in this work. We also evaluate the free energies of direct GB-protein interactions. The energetic properties of GB-protein association appear to scale with the size of the protein. For all proteins, the highly favorable change in free energy associated with direct protein-cosolvent interactions is nearly compensated by an unfavorable free energy of cavity formation (excluded volume effect), yielding a modestly unfavorable free energy for the transfer of a protein from water to a GB/water mixture.

  13. Typing of Typhoidal Salmonella Using Extraction of Water Soluble Whole Cell Proteins and Analysing by SDS-PAGE

    Directory of Open Access Journals (Sweden)

    R. Yousefi Mashouf

    2005-10-01

    Full Text Available Introduction & Objective : Salmonella is one of the most important genus of Enterobacteriacea family. The aim of this study was typing of typhoidal Salmonella by SDS-PAGE and comparing the results with those of serotyping method.Materials and Methods: In this study, 4 reference strains of Salmonella species, 5 reference strains of Enterobacteriacea family and 100 clinical isolates of Salmonella that were previously collected from laboratories of Hamadan medical centers were studied. Serotyping of strains were performed by Biomereux and Difco monovalent antisera. Whole-cell proteins of strains were also separated on 10% poly acrylamide gel. Gels were stained by Coomassie Brilliant Blue and analyzed by densitometry. Results: Of 100 cases of Salmonella species, 43 cases (43% were S. typhi, 20 cases (20% were S. typhymurium, 12 cases (12% were S. para typhi B, 10 cases (10% were S. para typhi C, S. para typhi A 1 case (1% and other cases were non-typhoidal Salmonella. The results of serotyping were compared with the results obtained by SDS-PAGE. Many protein bands from 220 KDa to 18.5 KDa were detected by SDS-PAGE and they were used to differentiate the strains. S. typhi serotypes were divided into 5 sub-species and S. para typhi B and C were divided each into 3 sub-species. Protein profiles of the reference strains of Salmonella were compared with protein profiles of Enterobacteriaceae species and showed some differences in major protein bands, however, they had a very similar protein band in 43 KDa area. Conclusion: Since our data was able to divide Salmonella species to sub-types and differentiate them from Enterobacteriacea species, we concluded that analsying SDS-PAGE profile of water soluble whole-cell proteins can be used for typing of these organisms and it is comparble with serotyping, nevertheless, further researches are needed to establish SDS-PAGE method and to replace it with serotyping method.

  14. Imprint of galaxy formation and evolution on globular cluster properties

    OpenAIRE

    Bekki, Kenji

    2006-01-01

    We discuss the origin of physical properties of globular cluster systems (GCSs) in galaxies in terms of galaxy formation and evolution processes. Based on numerical simulations of dynamical evolution of GCSs in galaxies, we particularly discuss (1) the origin of radial density profiles of GCSs, (2) kinematics of GCSs in elliptical galaxies, (3) transformation from nucleated dwarf galaxies into GCs (e.g., omega Centauri), and (4) the origin of GCSs in the Large Magellanic Cloud (LMC).

  15. Protein-Protein Docking in Drug Design and Discovery.

    Science.gov (United States)

    Kaczor, Agnieszka A; Bartuzi, Damian; Stępniewski, Tomasz Maciej; Matosiuk, Dariusz; Selent, Jana

    2018-01-01

    Protein-protein interactions (PPIs) are responsible for a number of key physiological processes in the living cells and underlie the pathomechanism of many diseases. Nowadays, along with the concept of so-called "hot spots" in protein-protein interactions, which are well-defined interface regions responsible for most of the binding energy, these interfaces can be targeted with modulators. In order to apply structure-based design techniques to design PPIs modulators, a three-dimensional structure of protein complex has to be available. In this context in silico approaches, in particular protein-protein docking, are a valuable complement to experimental methods for elucidating 3D structure of protein complexes. Protein-protein docking is easy to use and does not require significant computer resources and time (in contrast to molecular dynamics) and it results in 3D structure of a protein complex (in contrast to sequence-based methods of predicting binding interfaces). However, protein-protein docking cannot address all the aspects of protein dynamics, in particular the global conformational changes during protein complex formation. In spite of this fact, protein-protein docking is widely used to model complexes of water-soluble proteins and less commonly to predict structures of transmembrane protein assemblies, including dimers and oligomers of G protein-coupled receptors (GPCRs). In this chapter we review the principles of protein-protein docking, available algorithms and software and discuss the recent examples, benefits, and drawbacks of protein-protein docking application to water-soluble proteins, membrane anchoring and transmembrane proteins, including GPCRs.

  16. Intracellular protein breakdown. 8

    International Nuclear Information System (INIS)

    Bohley, P.; Kirschke, H.; Langner, J.; Wiederanders, B.; Ansorge, S.

    1976-01-01

    Double-labelled proteins from rat liver cytosol ( 14 C in long-lived, 3 H in short-lived proteins after in-vivo-labelling) are used as substrates for unlabelled proteinases in vitro. Differences in the degradation rates of short-lived and long-lived proteins in vitro by different proteinases and after addition of different effectors allow conclusions concerning their importance for the in-vivo-turnover of substrate proteins. The main activity (>90%) of soluble lysosomal proteinases at pH 6.1 and pH 6.9 is caused by thiolproteinases, which degrade preferentially short-lived cytosol proteins. These proteinases are inhibited by leupeptin. Autolysis of double-labelled cell fractions shows a remarkably faster breakdown of short-lived substrate proteins only in the soluble part of lysosomes. Microsomal fractions degrade in vitro preferentially long-lived substrate proteins. (author)

  17. Understanding the central kinematics of globular clusters with simulated integrated-light IFU observations

    Science.gov (United States)

    Bianchini, Paolo; Norris, Mark A.; van de Ven, Glenn; Schinnerer, Eva

    2015-10-01

    The detection of intermediate-mass black holes in the centres of globular clusters is highly controversial, as complementary observational methods often deliver significantly different results. In order to understand these discrepancies, we develop a procedure to simulate integral field unit (IFU) observations of globular clusters: Simulating IFU Star Cluster Observations (SISCO). The inputs of our software are realistic dynamical models of globular clusters that are then converted in a spectral data cube. We apply SISCO to Monte Carlo cluster simulations with a realistic number of stars and concentrations. Using independent realizations of a given simulation we are able to quantify the stochasticity intrinsic to the problem of observing a partially resolved stellar population with integrated-light spectroscopy. We show that the luminosity-weighted IFU observations can be strongly biased by the presence of a few bright stars that introduce a scatter in the velocity dispersion measurements up to ≃40 per cent around the expected value, preventing any sound assessment of the central kinematic and a sensible interpretation of the presence/absence of an intermediate-mass black hole. Moreover, we illustrate that, in our mock IFU observations, the average kinematic tracer has a mass of ≃0.75 M⊙, only slightly lower than the mass of the typical stars examined in studies of resolved line-of-sight velocities of giant stars. Finally, in order to recover unbiased kinematic measurements we test different masking techniques that allow us to remove the spaxels dominated by bright stars, bringing the scatter down to a level of only a few per cent. The application of SISCO will allow us to investigate state-of-the-art simulations as realistic observations.

  18. Hubble Space Telescope Observations of cD Galaxies and Their Globular Cluster Systems

    Science.gov (United States)

    Jordán, Andrés; Côté, Patrick; West, Michael J.; Marzke, Ronald O.; Minniti, Dante; Rejkuba, Marina

    2004-01-01

    We have used WFPC2 on the Hubble Space Telescope (HST) to obtain F450W and F814W images of four cD galaxies (NGC 541 in Abell 194, NGC 2832 in Abell 779, NGC 4839 in Abell 1656, and NGC 7768 in Abell 2666) in the range 5400 km s-1cluster (GC) systems reveals no anomalies in terms of specific frequencies, metallicity gradients, average metallicities, or the metallicity offset between the globular clusters and the host galaxy. We show that the latter offset appears roughly constant at Δ[Fe/H]~0.8 dex for early-type galaxies spanning a luminosity range of roughly 4 orders of magnitude. We combine the globular cluster metallicity distributions with an empirical technique described in a series of earlier papers to investigate the form of the protogalactic mass spectrum in these cD galaxies. We find that the observed GC metallicity distributions are consistent with those expected if cD galaxies form through the cannibalism of numerous galaxies and protogalactic fragments that formed their stars and globular clusters before capture and disruption. However, the properties of their GC systems suggest that dynamical friction is not the primary mechanism by which these galaxies are assembled. We argue that cD's instead form rapidly, via hierarchical merging, prior to cluster virialization. Based on observations with the NASA/ESA Hubble Space Telescope obtained at the Space Telescope Science Institute, which is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS 5-26555 Based in part on observations obtained at the European Southern Observatory, for VLT program 68.D-0130(A).

  19. New cataclysmic variables and other exotic binaries in the globular cluster 47 Tucanae*

    Science.gov (United States)

    Rivera Sandoval, L. E.; van den Berg, M.; Heinke, C. O.; Cohn, H. N.; Lugger, P. M.; Anderson, J.; Cool, A. M.; Edmonds, P. D.; Wijnands, R.; Ivanova, N.; Grindlay, J. E.

    2018-04-01

    We present 22 new (+3 confirmed) cataclysmic variables (CVs) in the non-core-collapsed globular cluster 47 Tucanae (47 Tuc). The total number of CVs in the cluster is now 43, the largest sample in any globular cluster so far. For the identifications we used near-ultraviolet (NUV) and optical images from the Hubble Space Telescope, in combination with X-ray results from the Chandra X-ray Observatory. This allowed us to build the deepest NUV CV luminosity function of the cluster to date. We found that the CVs in 47 Tuc are more concentrated towards the cluster centre than the main-sequence turn-off stars. We compared our results to the CV populations of the core-collapsed globular clusters NGC 6397 and NGC 6752. We found that 47 Tuc has fewer bright CVs per unit mass than those two other clusters. That suggests that dynamical interactions in core-collapsed clusters play a major role creating new CVs. In 47 Tuc, the CV population is probably dominated by primordial and old dynamically formed systems. We estimated that the CVs in 47 Tuc have total masses of ˜1.4 M⊙. We also found that the X-ray luminosity function of the CVs in the three clusters is bimodal. Additionally, we discuss a possible double degenerate system and an intriguing/unclassified object. Finally, we present four systems that could be millisecond pulsar companions given their X-ray and NUV/optical colours. For one of them we present very strong evidence for being an ablated companion. The other three could be CO or He white dwarfs.

  20. Cloning and expression of adiponectin and its globular domain, and measurement of the biological activity in vivo.

    Science.gov (United States)

    Hu, Xiao-Bo; Zhang, Yu-Jian; Zhang, Hui-Tang; Yang, Sheng-Li; Gong, Yi

    2003-11-01

    3T3-L1-adipocytes produce the adipocyte complement related protein of 30 kD (ACRP30), which is exclusively expressed in differentiated adipocytes. Decreased expression of ACRP30 correlates with insulin resistance in mouse models of altered insulin sensitivity. Adiponectin, the human homologue of ACRP30, circulates in human plasma at high levels. Plasma adiponectin levels have been reported to be decreased in some insulin-resistant states, such as obesity and type II diabetes mellitus. Here, full-length adiponectin and its C-terminal globular head domain (gAdiponectin) were expressed in Escherichia coli and gAdiponectin was used to immunize a rabbit to obtain polyclonal antiserum with titer of 10,000. Adiponectin was detected in human plasma with the use of gAdiponectin anti-serum by Western blot analysis, which was also detected by gACRP30 anti-serum. Injection in alloxan-treated rats with purified recombinant fusion adiponectin or gAdiponectin transiently abolished hyperglycemia. So adiponectin and gAdiponectin might have activity as a glucose lowering agent and potentially as a therapeutic for metabolic disease. All these results suggested that the recombinant protein had biological activity, and provided a useful tool in further studies.