WorldWideScience

Sample records for lysine residues modified

  1. Computational prediction of methylation types of covalently modified lysine and arginine residues in proteins.

    Science.gov (United States)

    Deng, Wankun; Wang, Yongbo; Ma, Lili; Zhang, Ying; Ullah, Shahid; Xue, Yu

    2016-05-30

    Protein methylation is an essential posttranslational modification (PTM) mostly occurs at lysine and arginine residues, and regulates a variety of cellular processes. Owing to the rapid progresses in the large-scale identification of methylation sites, the available data set was dramatically expanded, and more attention has been paid on the identification of specific methylation types of modification residues. Here, we briefly summarized the current progresses in computational prediction of methylation sites, which provided an accurate, rapid and efficient approach in contrast with labor-intensive experiments. We collected 5421 methyllysines and methylarginines in 2592 proteins from the literature, and classified most of the sites into different types. Data analyses demonstrated that different types of methylated proteins were preferentially involved in different biological processes and pathways, whereas a unique sequence preference was observed for each type of methylation sites. Thus, we developed a predictor of GPS-MSP, which can predict mono-, di- and tri-methylation types for specific lysines, and mono-, symmetric di- and asymmetrical di-methylation types for specific arginines. We critically evaluated the performance of GPS-MSP, and compared it with other existing tools. The satisfying results exhibited that the classification of methylation sites into different types for training can considerably improve the prediction accuracy. Taken together, we anticipate that our study provides a new lead for future computational analysis of protein methylation, and the prediction of methylation types of covalently modified lysine and arginine residues can generate more useful information for further experimental manipulation.

  2. Differential Contributions of Ubiquitin-Modified APOBEC3G Lysine Residues to HIV-1 Vif-Induced Degradation.

    Science.gov (United States)

    Turner, Tiffany; Shao, Qiujia; Wang, Weiran; Wang, Yudi; Wang, Chenliang; Kinlock, Ballington; Liu, Bindong

    2016-08-28

    Apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (A3G) is a host restriction factor that impedes HIV-1 replication. Viral integrity is salvaged by HIV-1 virion infectivity factor (Vif), which mediates A3G polyubiquitination and subsequent cellular depletion. Previous studies have implied that A3G polyubiquitination is essential for Vif-induced degradation. However, the contribution of polyubiquitination to the rate of A3G degradation remains unclear. Here, we show that A3G polyubiquitination is essential for degradation. Inhibition of ubiquitin-activating enzyme E1 by PYR-41 or blocking the formation of ubiquitin chains by over-expressing the lysine to arginine mutation of ubiquitin K48 (K48R) inhibited A3G degradation. Our A3G mutagenesis study showed that lysine residues 297, 301, 303, and 334 were not sufficient to render lysine-free A3G sensitive to Vif-mediated degradation. Our data also confirm that Vif could induce ubiquitin chain formation on lysine residues interspersed throughout A3G. Notably, A3G degradation relied on the lysine residues involved in polyubiquitination. Although A3G and the A3G C-terminal mutant interacted with Vif and were modified by ubiquitin chains, the latter remained more resistant to Vif-induced degradation. Furthermore, the A3G C-terminal mutant, but not the N-terminal mutant, maintained potent antiviral activity in the presence of Vif. Taken together, our results suggest that the location of A3G ubiquitin modification is a determinant for Vif-mediated degradation, implying that in addition to polyubiquitination, other factors may play a key role in the rate of A3G degradation.

  3. The structural feature surrounding glycated lysine residues in human hemoglobin.

    Science.gov (United States)

    Ito, Shigenori; Nakahari, Takashi; Yamamoto, Daisuke

    2011-06-01

    Complications derived from diabetes mellitus are caused by nonenzymatic protein glycation at the specific sites. LC/MS/MS was performed for the identification of the tryptic peptides of glycated hemoglobins using glyceraldehyde. After the identification of the glycation or non-glycation site, computer analysis of the structure surrounding the sites was carried out using PDB data (1BZ0). Five glycated lysine residues (Lys-16(α), -56(α), -8(β), -82(β), and -144(β)) and four non-glycated lysine residues (Lys-7(α), -40(α), -99(α), and -132(β)) were identified. The non-glycated lysine residues, Lys-7(α), -40(α), and -132(β), are most likely to form electrostatic interactions with the β carboxyl group of Asp-74(α), C-terminal His-146(β), and Glu-7(β) by virtue of their proximity, which is 2.67-2.91 Å (N-O). Additionally, there are histidine residues within 4.55-7.38 Å (N-N) around eight sites except for Lys-7(α). We conclude that the following factors seem to be necessary for glycation of lysine residues: (i) the apparent absence of aspartate or glutamate residues to inhibit the glycation reaction by forming an electrostatic interaction, (ii) the presence of histidine residues for acid-base catalysis of the Amadori rearrangement, and (iii) the presence of an amino acid residue capable of stabilizing a phosphate during proton transfer.

  4. Studies on the biotin-binding site of avidin. Lysine residues involved in the active site.

    Science.gov (United States)

    Gitlin, G; Bayer, E A; Wilchek, M

    1987-01-01

    Egg-white avidin was treated with 1-fluoro-2,4-dinitrobenzene. Modification of an average of one lysine residue per avidin subunit caused the complete loss of biotin binding. Tryptic peptides obtained from the 2,4-dinitrophenylated avidin were fractionated by reversed-phase h.p.l.c. Three peptides contained the 2,4-dinitrophenyl group. Amino acid analysis revealed that lysine residues 45, 94 and 111 are modified and probably comprise part of the biotin-binding site. PMID:3109401

  5. Studies on the biotin-binding site of avidin. Lysine residues involved in the active site.

    OpenAIRE

    Gitlin, G; Bayer, E A; Wilchek, M

    1987-01-01

    Egg-white avidin was treated with 1-fluoro-2,4-dinitrobenzene. Modification of an average of one lysine residue per avidin subunit caused the complete loss of biotin binding. Tryptic peptides obtained from the 2,4-dinitrophenylated avidin were fractionated by reversed-phase h.p.l.c. Three peptides contained the 2,4-dinitrophenyl group. Amino acid analysis revealed that lysine residues 45, 94 and 111 are modified and probably comprise part of the biotin-binding site.

  6. Role of lysine and acidic amino acid residues on the insecticidal activity of Jackbean urease.

    Science.gov (United States)

    Real-Guerra, Rafael; Carlini, Célia Regina; Stanisçuaski, Fernanda

    2013-09-01

    Canavalia ensiformis has three isoforms of urease: Jackbean urease (JBU), Jackbean urease II and canatoxin. These isoforms present several biological activities, independent from the enzymatic property, such as entomotoxicity and antifungal properties. The entomotoxic activity is a property of the whole protein, as well as of a 10 kDa peptide released by insect digestive enzymes. Here we have used chemical modification to observe the influence of lysines and acidic residues on JBU enzymatic and insecticidal activities. Chemical modification of lysine residues was performed with dimethylamine-borane complex and formaldehyde, and acidic residues were modified by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and ethylenediamine. Derivatized ureases, called JBU-Lys (lysine-modified) and JBU-Ac (acidic residues-modified), were assayed for their biochemical and insecticidal properties. Neither modification altered significantly the kinetic parameters analyzed, indicating that no residue critical for the enzyme activity was affected and that the modifications did not incur in any significant structural alteration. On the other hand, both modifications reduced the toxic activity of the native protein fed to Dysdercus peruvianus. The changes observed in the entomotoxic property of the derivatized proteins reflect alterations in different steps of JBU's toxicity towards insects. JBU-Ac is not susceptible to hydrolysis by insect digestive enzymes, hence impairing the release of toxic peptide(s), while JBU-Lys is processed as the native protein. On the other hand, the antidiuretic effect of JBU on Rhodnius prolixus is altered in JBU-Lys, but not in JBU-Ac. Altogether, these data emphasize the role of lysine and acidic residues on the insecticidal properties of ureases.

  7. Antibacterial activity of a newly developed peptide-modified lysin against Acinetobacter baumannii and Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Hang eYang

    2015-12-01

    Full Text Available The global emergence of multidrug-resistant (MDR bacteria is a growing threat to public health worldwide. Natural bacteriophage lysins are promising alternatives in the treatment of infections caused by Gram-positive pathogens, but not Gram-negative ones, like Acinetobacter baumannii and Pseudomonas aeruginosa, due to the barriers posed by their outer membranes. Recently, modifying a natural lysin with an antimicrobial peptide was found able to break the barriers, and to kill Gram-negative pathogens. Herein, a new peptide-modified lysin (PlyA was constructed by fusing the cecropin A peptide residues 1–8 (KWKLFKKI with the OBPgp279 lysin and its antibacterial activity was studied. PlyA showed good and broad antibacterial activities against logarithmic phase A. baumannii and P. aeruginosa, but much reduced activities against the cells in stationary phase. Addition of outer membrane permeabilizers (EDTA and citric acid could enhance the antibacterial activity of PlyA against stationary phase cells. Finally, no antibacterial activity of PlyA could be observed in some bio-matrices, such as culture media, milk, and sera. In conclusion, we reported here a novel peptide-modified lysin with significant antibacterial activity against both logarithmic (without OMPs and stationary phase (with OMPs A. baumannii and P. aeruginosa cells in buffer, but further optimization is needed to achieve broad activity in diverse bio-matrices.

  8. Lysine residue 185 of Rad1 is a topological but not a functional counterpart of lysine residue 164 of PCNA.

    Directory of Open Access Journals (Sweden)

    Niek Wit

    Full Text Available Monoubiquitylation of the homotrimeric DNA sliding clamp PCNA at lysine residue 164 (PCNA(K164 is a highly conserved, DNA damage-inducible process that is mediated by the E2/E3 complex Rad6/Rad18. This ubiquitylation event recruits translesion synthesis (TLS polymerases capable of replicating across damaged DNA templates. Besides PCNA, the Rad6/Rad18 complex was recently shown in yeast to ubiquitylate also 9-1-1, a heterotrimeric DNA sliding clamp composed of Rad9, Rad1, and Hus1 in a DNA damage-inducible manner. Based on the highly similar crystal structures of PCNA and 9-1-1, K185 of Rad1 (Rad1(K185 was identified as the only topological equivalent of PCNA(K164. To investigate a potential role of posttranslational modifications of Rad1(K185 in DNA damage management, we here generated a mouse model with a conditional deletable Rad1(K185R allele. The Rad1(K185 residue was found to be dispensable for Chk1 activation, DNA damage survival, and class switch recombination of immunoglobulin genes as well as recruitment of TLS polymerases during somatic hypermutation of immunoglobulin genes. Our data indicate that Rad1(K185 is not a functional counterpart of PCNA(K164.

  9. A noncanonical function of sortase enables site-specific conjugation of small molecules to lysine residues in proteins.

    Science.gov (United States)

    Bellucci, Joseph J; Bhattacharyya, Jayanta; Chilkoti, Ashutosh

    2015-01-07

    We provide the first demonstration that isopeptide ligation, a noncanonical activity of the enzyme sortase A, can be used to modify recombinant proteins. This reaction was used in vitro to conjugate small molecules to a peptide, an engineered targeting protein, and a full-length monoclonal antibody with an exquisite level of control over the site of conjugation. Attachment to the protein substrate occurred exclusively through isopeptide bonds at a lysine ε-amino group within a specific amino acid sequence. This reaction allows more than one molecule to be site-specifically conjugated to a protein at internal sites, thereby overcoming significant limitations of the canonical native peptide ligation reaction catalyzed by sortase A. Our method provides a unique chemical ligation procedure that is orthogonal to existing methods, supplying a new method to site-specifically modify lysine residues that will be a valuable addition to the protein conjugation toolbox.

  10. (R)-β-lysine-modified elongation factor P functions in translation elongation

    DEFF Research Database (Denmark)

    Bullwinkle, Tammy J; Zou, S Betty; Rajkovic, Andrei

    2013-01-01

    Post-translational modification of bacterial elongation factor P (EF-P) with (R)-β-lysine at a conserved lysine residue activates the protein in vivo and increases puromycin reactivity of the ribosome in vitro. The additional hydroxylation of EF-P at the same lysine residue by the YfcM protein has......-(β)-lysyl-EF-P showed 30% increased puromycin reactivity but no differences in dipeptide synthesis rates when compared with the β-lysylated form. Unlike disruption of the other genes required for EF-P modification, deletion of yfcM had no phenotypic consequences in Salmonella. Taken together, our findings indicate...

  11. The tRNA synthetase paralog PoxA modifies elongation factor-P with (R)-ß-lysine

    DEFF Research Database (Denmark)

    Roy, Hervé; Zou, S Betty; Bullwinkle, Tammy J

    2011-01-01

    The lysyl-tRNA synthetase paralog PoxA modifies elongation factor P (EF-P) with a-lysine at low efficiency. Cell-free extracts containing non-a-lysine substrates of PoxA modified EF-P with a change in mass consistent with addition of ß-lysine, a substrate also predicted by genomic analyses. EF-P ...

  12. Role of lysine binding residues in the global folding of the lysC riboswitch.

    Science.gov (United States)

    Smith-Peter, Erich; Lamontagne, Anne-Marie; Lafontaine, Daniel A

    2015-01-01

    Riboswitches regulate gene expression by rearranging their structure upon metabolite binding. The lysine-sensing lysC riboswitch is a rare example of an RNA aptamer organized around a 5-way helical junction in which ligand binding is performed exclusively through nucleotides located at the junction core. We have probed whether the nucleotides involved in ligand binding play any role in the global folding of the riboswitch. As predicted, our findings indicate that ligand-binding residues are critical for the lysine-dependent gene regulation mechanism. We also find that these residues are not important for the establishment of key magnesium-dependent tertiary interactions, suggesting that folding and ligand recognition are uncoupled in this riboswitch for the formation of specific interactions. However, FRET assays show that lysine binding results in an additional conformational change, indicating that lysine binding may also participate in a specific folding transition. Thus, in contrast to helical junctions being primary determinants in ribozymes and rRNA folding, we speculate that the helical junction of the lysine-sensing lysC riboswitch is not employed as structural a scaffold to direct global folding, but rather has a different role in establishing RNA-ligand interactions required for riboswitch regulation. Our work suggests that helical junctions may adopt different functions such as the coordination of global architecture or the formation of specific ligand binding site.

  13. Regulation of translesion DNA synthesis: Posttranslational modification of lysine residues in key proteins.

    Science.gov (United States)

    McIntyre, Justyna; Woodgate, Roger

    2015-05-01

    Posttranslational modification of proteins often controls various aspects of their cellular function. Indeed, over the past decade or so, it has been discovered that posttranslational modification of lysine residues plays a major role in regulating translesion DNA synthesis (TLS) and perhaps the most appreciated lysine modification is that of ubiquitination. Much of the recent interest in ubiquitination stems from the fact that proliferating cell nuclear antigen (PCNA) was previously shown to be specifically ubiquitinated at K164 and that such ubiquitination plays a key role in regulating TLS. In addition, TLS polymerases themselves are now known to be ubiquitinated. In the case of human polymerase η, ubiquitination at four lysine residues in its C-terminus appears to regulate its ability to interact with PCNA and modulate TLS. Within the past few years, advances in global proteomic research have revealed that many proteins involved in TLS are, in fact, subject to a previously underappreciated number of lysine modifications. In this review, we will summarize the known lysine modifications of several key proteins involved in TLS; PCNA and Y-family polymerases η, ι, κ and Rev1 and we will discuss the potential regulatory effects of such modification in controlling TLS in vivo.

  14. Critical lysine residues of Klf4 required for protein stabilization and degradation

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Key-Hwan; Kim, So-Ra; Ramakrishna, Suresh; Baek, Kwang-Hyun, E-mail: baek@cha.ac.kr

    2014-01-24

    Highlights: • Klf4 undergoes the 26S proteasomal degradation by ubiquitination on its multiple lysine residues. • Essential Klf4 ubiquitination sites are accumulated between 190–263 amino acids. • A mutation of lysine at 232 on Klf4 elongates protein turnover. • Klf4 mutants dramatically suppress p53 expression both under normal and UV irradiated conditions. - Abstract: The transcription factor, Krüppel-like factor 4 (Klf4) plays a crucial role in generating induced pluripotent stem cells (iPSCs). As the ubiquitination and degradation of the Klf4 protein have been suggested to play an important role in its function, the identification of specific lysine sites that are responsible for protein degradation is of prime interest to improve protein stability and function. However, the molecular mechanism regulating proteasomal degradation of the Klf4 is poorly understood. In this study, both the analysis of Klf4 ubiquitination sites using several Klf4 deletion fragments and bioinformatics predictions showed that the lysine sites which are signaling for Klf4 protein degradation lie in its N-terminal domain (aa 1–296). The results also showed that Lys32, 52, 232, and 252 of Klf4 are responsible for the proteolysis of the Klf4 protein. These results suggest that Klf4 undergoes proteasomal degradation and that these lysine residues are critical for Klf4 ubiquitination.

  15. Chemoselective small molecules that covalently modify one lysine in a non-enzyme protein in plasma

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Sungwook; Connelly, Stephen; Reixach, Natàlia; Wilson, Ian A.; Kelly, Jeffery W. (Scripps)

    2010-02-19

    A small molecule that could bind selectively to and then react chemoselectively with a non-enzyme protein in a complex biological fluid, such as blood, could have numerous practical applications. Herein, we report a family of designed stilbenes that selectively and covalently modify the prominent plasma protein transthyretin in preference to more than 4,000 other human plasma proteins. They react chemoselectively with only one of eight lysine {epsilon}-amino groups within transthyretin. The crystal structure confirms the expected binding orientation of the stilbene substructure and the anticipated conjugating amide bond. These covalent transthyretin kinetic stabilizers exhibit superior amyloid inhibition potency compared to their noncovalent counterparts, and they prevent cytotoxicity associated with amyloidogenesis. Though there are a few prodrugs that, upon metabolic activation, react with a cysteine residue inactivating a specific non-enzyme, we are unaware of designed small molecules that react with one lysine {epsilon}-amine within a specific non-enzyme protein in a complex biological fluid.

  16. SucStruct: Prediction of succinylated lysine residues by using structural properties of amino acids.

    Science.gov (United States)

    López, Yosvany; Dehzangi, Abdollah; Lal, Sunil Pranit; Taherzadeh, Ghazaleh; Michaelson, Jacob; Sattar, Abdul; Tsunoda, Tatsuhiko; Sharma, Alok

    2017-03-28

    Post-Translational Modification (PTM) is a biological reaction which contributes to diversify the proteome. Despite many modifications with important roles in the cellular activity, lysine succinylation has recently emerged as an important PTM mark. It alters the chemical structure of lysines, leading to remarkable changes in the structure and function of proteins. Given the huge amount of proteins being sequenced in the post-genome era, the experimental detection of succinylated residues remains expensive, inefficient and time-consuming. Therefore, the development of computational tools for accurately predicting succinylated lysines is an urgent necessity. To date, several approaches have been proposed but their sensitivity has been reportedly poor. In this paper, we propose an approach that utilizes structural features of amino acids to improve lysine succinylation prediction. Succinylated and non-succinylated lysines were first retrieved from 670 proteins and characteristics such as accessible surface area, backbone torsion angles, and local structure conformations were incorporated. We used the k-nearest neighbors cleaning for dealing with class imbalance and designed a pruned decision tree for classification. Our predictor, referred as SucStruct (Succinylation using Structural features), proved to significantly improve performance when compared to previous predictors, with sensitivity, accuracy and Mathew's correlation coefficient equal to 0.7334-0.7946, 0.7444-0.7608 and 0.4884-0.5240, respectively.

  17. Bioconjugation of Oligodeoxynucleotides Carrying 1,4-Dicarbonyl Groups via Reductive Amination with Lysine Residues.

    Science.gov (United States)

    Yang, Bo; Jinnouchi, Akiko; Usui, Kazuteru; Katayama, Tsutomu; Fujii, Masayuki; Suemune, Hiroshi; Aso, Mariko

    2015-08-19

    We evaluated the efficacy of bioconjugation of oligodeoxynucleotides (ODNs) containing 1,4-dicarbonyl groups, a C4'-oxidized abasic site (OAS), and a newly designed 2'-methoxy analogue, via reductive amination with lysine residues. Dicarbonyls, aldehyde and ketone at C1- and C4-positions of deoxyribose in the ring-opened form of OAS allowed efficient reaction with amines. Kinetic studies indicated that reductive amination of OAS-containing ODNs with a proximal amine on the complementary strand proceeded 10 times faster than the corresponding reaction of an ODN containing an abasic site with C1-aldehyde. Efficient reductive amination between the DNA-binding domain of Escherichia coli DnaA protein and ODNs carrying OAS in the DnaA-binding sequence proceeded at the lysine residue in proximity to the phosphate group at the 5'-position of the OAS, in contrast to unsuccessful conjugation with abasic site ODNs, even though they have similar aldehydes. Theoretical calculation indicated that the C1-aldehyde of OAS was more accessible to the target lysine than that of the abasic site. These results demonstrate the potential utility of cross-linking strategies that use dicarbonyl-containing ODNs for the study of protein-nucleic acid interactions. Conjugation with a lysine-containing peptide that lacked specific affinity for ODN was also successful, further highlighting the advantages of 1,4-dicarbonyls.

  18. Synthesis, pharmacokinetics, and biological use of lysine-modified single-walled carbon nanotubes.

    Science.gov (United States)

    Mulvey, J Justin; Feinberg, Evan N; Alidori, Simone; McDevitt, Michael R; Heller, Daniel A; Scheinberg, David A

    2014-01-01

    We aimed to create a more robust and more accessible standard for amine-modifying single-walled carbon nanotubes (SWCNTs). A 1,3-cycloaddition was developed using an azomethine ylide, generated by reacting paraformaldehyde and a side-chain-Boc (tert-Butyloxycarbonyl)-protected, lysine-derived alpha-amino acid, H-Lys(Boc)-OH, with purified SWCNT or C60. This cycloaddition and its lysine adduct provides the benefits of dense, covalent modification, ease of purification, commercial availability of reagents, and pH-dependent solubility of the product. Subsequently, SWCNTs functionalized with lysine amine handles were covalently conjugated to a radiometalated chelator, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA). The (111)In-labeled construct showed rapid renal clearance in a murine model and a favorable biodistribution, permitting utility in biomedical applications. Functionalized SWCNTs strongly wrapped small interfering RNA (siRNA). In the first disclosed deployment of thermophoresis with carbon nanotubes, the lysine-modified tubes showed a desirable, weak SWCNT-albumin binding constant. Thus, lysine-modified nanotubes are a favorable candidate for medicinal work.

  19. Effects of lysine residues on structural characteristics and stability of tau proteins

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Myeongsang; Baek, Inchul; Choi, Hyunsung; Kim, Jae In; Na, Sungsoo, E-mail: nass@korea.ac.kr

    2015-10-23

    Pathological amyloid proteins have been implicated in neuro-degenerative diseases, specifically Alzheimer's, Parkinson's, Lewy-body diseases and prion related diseases. In prion related diseases, functional tau proteins can be transformed into pathological agents by environmental factors, including oxidative stress, inflammation, Aβ-mediated toxicity and covalent modification. These pathological agents are stable under physiological conditions and are not easily degraded. This un-degradable characteristic of tau proteins enables their utilization as functional materials to capturing the carbon dioxides. For the proper utilization of amyloid proteins as functional materials efficiently, a basic study regarding their structural characteristic is necessary. Here, we investigated the basic tau protein structure of wild-type (WT) and tau proteins with lysine residues mutation at glutamic residue (Q2K) on tau protein at atomistic scale. We also reported the size effect of both the WT and Q2K structures, which allowed us to identify the stability of those amyloid structures. - Highlights: • Lysine mutation effect alters the structure conformation and characteristic of tau. • Over the 15 layers both WT and Q2K models, both tau proteins undergo fractions. • Lysine mutation causes the increment of non-bonded energy and solvent accessible surface area. • Structural instability of Q2K model was proved by the number of hydrogen bonds analysis.

  20. Functional importance of motif I of pseudouridine synthases: mutagenesis of aligned lysine and proline residues.

    Science.gov (United States)

    Spedaliere, C J; Hamilton, C S; Mueller, E G

    2000-08-01

    On the basis of sequence alignments, the pseudouridine synthases were grouped into four families that share no statistically significant global sequence similarity, though some common sequence motifs were discovered [Koonin, E. V. (1996) Nucleic Acids. Res. 24, 2411-2415; Gustafsson, C., Reid, R., Greene, P. J., and Santi, D. V. (1996) Nucleic Acids Res. 24, 3756-3762]. We have investigated the functional significance of these alignments by substituting the nearly invariant lysine and proline residues in Motif I of RluA and TruB, pseudouridine synthases belonging to different families. Contrary to our expectations, the altered enzymes display only very mild kinetic impairment. Substitution of the aligned lysine and proline residues does, however, reduce structural stability, consistent with a temperature sensitive phenotype that results from substitution of the cognate proline residue in Cbf5p, a yeast homologue of TruB [Zerbarjadian, Y., King, T., Fournier, M. J., Clarke, L., and Carbon, J. (1999) Mol. Cell. Biol. 19, 7461-7472]. Together, our data support a functional role for Motif I, as predicted by sequence alignments, though the effect of substituting the highly conserved residues was milder than we anticipated. By extrapolation, our findings also support the assignment of pseudouridine synthase function to certain physiologically important eukaryotic proteins that contain Motif I, including the human protein dyskerin, alteration of which leads to the disease dyskeratosis congenita.

  1. Structure-function relationships in scorpion neurotoxins. Identification of the supperreactive lysine residue in toxin I of Androctonus australis Hector.

    Science.gov (United States)

    Sampieri, F; Habersetzer-Rochat, C

    1978-07-21

    In a previous article (Habersetzer-Rochat, C. and Sampieri, R. (1976) Biochemistry 15, 2254--2261) it was demonstrated that the toxin I of the North African Scorpion Androctonus australis Hector was inactivated after reaction with iodoacetate; the toxicity loss in mice was correlated with the carboxymethylation of one superreactive residue. In the present work, alkylation of toxin I was performed with iodo[14C]-acetate. Hence, it was possible, after reduction, S-methylation and chymotryptic hydrolysis of this toxin, to isolate the peptide containing the labelled lysine residue. By automatic Edman degradation, this residue was identified as being the penultimate lysine at position 56 in the primary sequence. Comparison of three primary structures of scorpion neurotoxins and comparison in different kinds of activity seem to indicate that this lysine residue is mainly important for toxicity in mice.

  2. Superoxide reduction by a superoxide reductase lacking the highly conserved lysine residue.

    Science.gov (United States)

    Pinto, Ana F; Romão, Célia V; Pinto, Liliana C; Huber, Harald; Saraiva, Lígia M; Todorovic, Smilja; Cabelli, Diane; Teixeira, Miguel

    2015-01-01

    Superoxide reductases (SORs) are the most recently identified superoxide detoxification systems, being found in microorganisms from the three domains of life. These enzymes are characterized by a catalytic mononuclear iron site, with one cysteine and four histidine ligands of the ferrous active form. A lysine residue in the -EKHVP- motif, located close to the active site, has been considered to be essential for the enzyme function, by contributing to the positive surface patch that attracts the superoxide anion and by controlling the chemistry of the catalytic mechanism through a hydrogen bond network. However, we show here that this residue is substituted by non-equivalent amino acids in several putative SORs from Archaea and unicellular Eukarya. In this work, we focus on mechanistic and spectroscopic studies of one of these less common enzymes, the SOR from the hyperthermophilic Crenarchaeon Ignicoccus hospitalis. We employ pulse radiolysis fast kinetics and spectroscopic approaches to study the wild-type enzyme (-E23T24HVP-), and two mutants, T24K and E23A, the later mimicking enzymes lacking both the lysine and glutamate (a ferric ion ligand) of the motif. The efficiency of the wild-type protein and mutants in reducing superoxide is comparable to other SORs, revealing the robustness of these enzymes to single mutations.

  3. Synthesis, pharmacokinetics, and biological use of lysine-modified single-walled carbon nanotubes

    Directory of Open Access Journals (Sweden)

    Mulvey JJ

    2014-09-01

    Full Text Available J Justin Mulvey,1,2 Evan N Feinberg,1,3 Simone Alidori,1 Michael R McDevitt,4,5 Daniel A Heller,1,6 David A Scheinberg1,5,6 1Molecular Pharmacology and Chemistry Program, Sloan Kettering Institute, New York, NY, USA; 2Tri-Institutional MD-PhD Program, New York, NY, USA; 3Department of Applied Physics, Yale University, New Haven, CT USA; 4Department of Radiology and 5Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, USA; 6Weill Cornell Medical College, New York, NY, USA Abstract: We aimed to create a more robust and more accessible standard for amine-modifying single-walled carbon nanotubes (SWCNTs. A 1,3-cycloaddition was developed using an azomethine ylide, generated by reacting paraformaldehyde and a side-chain-Boc (tert-Butyloxycarbonyl-protected, lysine-derived alpha-amino acid, H-Lys(Boc-OH, with purified SWCNT or C60. This cycloaddition and its lysine adduct provides the benefits of dense, covalent modification, ease of purification, commercial availability of reagents, and pH-dependent solubility of the product. Subsequently, SWCNTs functionalized with lysine amine handles were covalently conjugated to a radiometalated chelator, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA. The 111In-labeled construct showed rapid renal clearance in a murine model and a favorable biodistribution, permitting utility in biomedical applications. Functionalized SWCNTs strongly wrapped small interfering RNA (siRNA. In the first disclosed deployment of thermophoresis with carbon nanotubes, the lysine-modified tubes showed a desirable, weak SWCNT-albumin binding constant. Thus, lysine-modified nanotubes are a favorable candidate for medicinal work. Keywords: fullerene, cycloaddition, azomethine, DOTA, thermophoresis, 111In

  4. The conserved Lysine69 residue plays a catalytic role in Mycobacterium tuberculosis shikimate dehydrogenase

    Directory of Open Access Journals (Sweden)

    Rodrigues Valnês

    2009-01-01

    Full Text Available Abstract Background The shikimate pathway is an attractive target for the development of antitubercular agents because it is essential in Mycobacterium tuberculosis, the causative agent of tuberculosis, but absent in humans. M. tuberculosis aroE-encoded shikimate dehydrogenase catalyzes the forth reaction in the shikimate pathway. Structural and functional studies indicate that Lysine69 may be involved in catalysis and/or substrate binding in M. tuberculosis shikimate dehydrogenase. Investigation of the kinetic properties of mutant enzymes can bring important insights about the role of amino acid residues for M. tuberculosis shikimate dehydrogenase. Findings We have performed site-directed mutagenesis, steady-state kinetics, equilibrium binding measurements and molecular modeling for both the wild-type M. tuberculosis shikimate dehydrogenase and the K69A mutant enzymes. The apparent steady-state kinetic parameters for the M. tuberculosis shikimate dehydrogenase were determined; the catalytic constant value for the wild-type enzyme (50 s-1 is 68-fold larger than that for the mutant K69A (0.73 s-1. There was a modest increase in the Michaelis-Menten constant for DHS (K69A = 76 μM; wild-type = 29 μM and NADPH (K69A = 30 μM; wild-type = 11 μM. The equilibrium dissociation constants for wild-type and K69A mutant enzymes are 32 (± 4 μM and 134 (± 21, respectively. Conclusion Our results show that the residue Lysine69 plays a catalytic role and is not involved in substrate binding for the M. tuberculosis shikimate dehydrogenase. These efforts on M. tuberculosis shikimate dehydrogenase catalytic mechanism determination should help the rational design of specific inhibitors, aiming at the development of antitubercular drugs.

  5. Inhibition of Alkaline Phosphatase from Pearl Oyster Pinctada fucata by o-Phthalaldehyde: Involvement of Lysine and Histidine Residues at the Active Site

    Institute of Scientific and Technical Information of China (English)

    CHEN Hongtao; XIE Liping; YU Zhenyan; ZHANG Rongqing

    2005-01-01

    Alkaline phosphatase from Pinctada fucata was inactivated by o-phthalaldehyde (OPA). The inactivation followed pseudo first-order kinetics with a second rate constant of 0.167 (mmol/L)-1·min-1 at pH 7.5 and 25°C. A Tsou's plot analysis showed that inactivation occurred upon formation of one isoindole group. The OPA-modified enzyme lost the ability to bind with the specific affinity column and the presence of substrates or competitive inhibitors protected the enzyme from inactivation. The results revealed that the OPA-reaction site was at the enzyme substrate binding site. Prior modification of the enzyme by lysine or histidine specific reagent abolished formation of the isoindole derivatives, suggesting that lysine and histidine residues were involved in the OPA-induced inactivation. Taken together, OPA inactivated the alkaline phosphatase from Pinctada fucata by cross-linking lysine and histidine residues at the active site and formed an isoindole group at the substrate binding site of the enzyme.

  6. Poly(L-lysine)-modified iron oxide nanoparticles for stem cell labeling.

    Science.gov (United States)

    Babic, Michal; Horák, Daniel; Trchová, Miroslava; Jendelová, Pavla; Glogarová, Katerina; Lesný, Petr; Herynek, Vít; Hájek, Milan; Syková, Eva

    2008-03-01

    New surface-modified iron oxide nanoparticles were developed by precipitation of Fe(II) and Fe(III) salts with ammonium hydroxide and oxidation of the resulting magnetite with sodium hypochlorite, followed by the addition of poly( L-lysine) (PLL) solution. PLL of several molecular weights ranging from 146 ( L-lysine) to 579 000 was tested as a coating to boost the intracellular uptake of the nanoparticles. The nanoparticles were characterized by TEM, dynamic light scattering, FTIR, and ultrasonic spectrometry. TEM revealed that the particles were ca. 6 nm in diameter, while FTIR showed that their surfaces were well-coated with PLL. The interaction of PLL-modified iron oxide nanoparticles with DMEM culture medium was verified by UV-vis spectroscopy. Rat bone marrow stromal cells (rMSCs) and human mesenchymal stem cells (hMSC) were labeled with PLL-modified iron oxide nanoparticles or with Endorem (control). Optical microscopy and TEM confirmed the presence of PLL-modified iron oxide nanoparticles inside the cells. Cellular uptake was very high (more than 92%) for PLL-modified nanoparticles that were coated with PLL (molecular weight 388 00) at a concentration of 0.02 mg PLL per milliliter of colloid. The cellular uptake of PLL-modified iron oxide was facilitated by its interaction with the negatively charged cell surface and subsequent endosomolytic uptake. The relaxivity of rMSCs labeled with PLL-modified iron oxide and the amount of iron in the cells were determined. PLL-modified iron oxide-labeled rMSCs were imaged in vitro and in vivo after intracerebral grafting into the contralateral hemisphere of the adult rat brain. The implanted cells were visible on magnetic resonance (MR) images as a hypointense area at the injection site and in the lesion. In comparison with Endorem, nanoparticles modified with PLL of an optimum molecular weight demonstrated a higher efficiency of intracellular uptake by MSC cells.

  7. Preparation of Lysine Modified Chitosan Nanoparticles%赖氨酸改性壳聚糖的制备研究

    Institute of Scientific and Technical Information of China (English)

    符思达; 沈逸敏; 陈巧玲; 朱培培; 汤汉; 王亨缇; 孙燕

    2012-01-01

    Lysine modified chitosan was prepared and characterized in the study. A series of reaction parameters, such as ratio of lysine and chitosan, were tested. The lysine grafted chitosan was characterized by fourier transform infrared spectroscopy and differential scanning calorimetry, and the water-soluable ability of lysine grafted chitosan also was tested. The results showed that lysine grafted chitosan was synthesized successfully and the optimal reaction parameters were as following: the ratio of lysine and chitosan was 3 : 1. After lysine modified, the water-soluable ability of chitosan was improved significantly.%本实验以赖氨酸改性壳聚糖为研究对象,对赖氨酸对壳聚糖的改性进行研究。改变壳聚糖与赖氨酸的质量比研究其改性效果。差示扫描量热分析、红外光谱表征以及水溶性试验表明赖氨酸改性壳聚糖是成功的。赖氨酸与壳聚糖的比例为3:1的时,改性较好。赖氨酸改性后的壳聚糖其水溶性显著提高。

  8. Lysine residues at the first and second KTKEGV repeats mediate α-Synuclein binding to membrane phospholipids.

    Science.gov (United States)

    Zarbiv, Yonaton; Simhi-Haham, Dganit; Israeli, Eitan; Elhadi, Suaad Abed; Grigoletto, Jessica; Sharon, Ronit

    2014-10-01

    While α-Synuclein (α-Syn) is mainly detected as a cytosolic protein, a portion of it is recovered bound to membranes. It is suggested that binding to membrane phospholipids controls α-Syn structure, physiology and pathogenesis. We aimed at investigating the role, of the positive charged lysine residues at the KTKEGV repeat motif, in mediating α-Syn associations with membrane phospholipids and in α-Syn oligomerization and aggregation. Specifically, two positive lysine (K) residues were replaced with two negative glutamic acid (E) residues at either the first or second KTKEGV repeat motifs. The effect of these mutations on membrane binding was determined by a quantitative phospholipid ELISA assay and compared to wild-type α-Syn and to the Parkinson's disease-causing mutations, A30P, E46K and A53T. We found that the K to E substitutions affected α-Syn binding to phospholipids. In addition, K to E substitutions resulted in a dramatically lower level of soluble α-Syn oligomers and larger intracellular inclusions. Together, our results suggest a critical role for lysine residues at the N-terminal repeat domain in the pathophysiology of α-Syn.

  9. Immobilization of lysine oxidase on a gold-platinum nanoparticles modified Au electrode for detection of lysine.

    Science.gov (United States)

    Chauhan, N; Narang, J; Sunny; Pundir, C S

    2013-04-10

    A commercial lysine oxidase (LyOx) from Trichoderma viride was immobilized covalently onto gold nanoparticles (AuNPs) and platinum nanoparticles (PtNPs) electrodeposited onto Au electrode using 3-aminopropyltriethoxy silane (3-APTES) and glutaraldehyde cross linking chemistry. A lysine biosensor was fabricated using LyOx/3-APTES/AuNPs-PtNPs/Au electrode as a working electrode, Ag/AgCl (3M KCl) as standard electrode and Pt wire as auxiliary electrode connected through a potentiostat. The enzyme electrode was characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). The cumulative effect of AuNPs and PtNPs showed excellent electrocatalytic activity at low applied potential for detection of H2O2, a product of LyOx reaction. The sensor showed its optimum response within 4s, when polarized at 0.2V vs. Ag/AgCl in 0.1M phosphate buffer, pH 7.5 at 30°C. The linear range and detection limit of the sensor were 1.0-600μM and 1.0μM (S/N=3), respectively. Biosensor measured lysine level in sera, milk and amino acid tablet, which correlated well with those by standard HPLC method. The enzyme electrode lost 50% of its initial activity after 200 uses over a period of 4 months.

  10. Elicitin-Induced Distal Systemic Resistance in Plants is Mediated Through the Protein-Protein Interactions Influenced by Selected Lysine Residues.

    Science.gov (United States)

    Uhlíková, Hana; Obořil, Michal; Klempová, Jitka; Šedo, Ondrej; Zdráhal, Zbyněk; Kašparovský, Tomáš; Skládal, Petr; Lochman, Jan

    2016-01-01

    Elicitins are a family of small proteins with sterol-binding activity that are secreted by Phytophthora and Pythium sp. classified as oomycete PAMPs. Although α- and β-elicitins bind with the same affinity to one high affinity binding site on the plasma membrane, β-elicitins (possessing 6-7 lysine residues) are generally 50- to 100-fold more active at inducing distal HR and systemic resistance than the α-isoforms (with only 1-3 lysine residues). To examine the role of lysine residues in elicitin biological activity, we employed site-directed mutagenesis to prepare a series of β-elicitin cryptogein variants with mutations on specific lysine residues. In contrast to direct infiltration of protein into leaves, application to the stem revealed a rough correlation between protein's charge and biological activity, resulting in protection against Phytophthora parasitica. A detailed analysis of proteins' movement in plants showed no substantial differences in distribution through phloem indicating differences in consequent apoplastic or symplastic transport. In this process, an important role of homodimer formation together with the ability to form a heterodimer with potential partner represented by endogenous plants LTPs is suggested. Our work demonstrates a key role of selected lysine residues in these interactions and stresses the importance of processes preceding elicitin recognition responsible for induction of distal systemic resistance.

  11. Elicitin-induced distal systemic resistance in plants is mediated through the protein-protein interactions influenced by selected lysine residues

    Directory of Open Access Journals (Sweden)

    Hana eUhlíková

    2016-02-01

    Full Text Available Elicitins are a family of small proteins with sterol-binding activity that are secreted by Phytophthora and Pythium spp. classified as oomycete PAMPs. Although alfa- and beta-elicitins bind with the same affinity to one high affinity binding site on the plasma membrane, beta-elicitins (possessing 6-7 lysine residues are generally 50- to 100-fold more active at inducing distal HR and systemic resistance than the alfa-isoforms (with only 1-3 lysine residues.To examine the role of lysine residues in elicitin biological activity, we employed site-directed mutagenesis to prepare a series of beta-elicitin cryptogein variants with mutations on specific lysine residues. In contrast to direct infiltration of protein into leaves, application to the stem revealed a rough correlation between protein’s charge and biological activity, resulting in protection against Phytophthora parasitica. A detailed analysis of proteins’ movement in plants showed no substantial differences in distribution through phloem indicating differences in consequent apoplastic or symplastic transport. In this process, an important role of homodimer formation together with the ability to form a heterodimer with potential partner represented by endogenous plants LTPs is suggested. Our work demonstrates a key role of selected lysine residues in these interactions and stresses the importance of processes preceding elicitin recognition responsible for induction of distal systemic resistance.

  12. Oxidative deamination of benzylamine and lysine residue in bovine serum albumin by green tea, black tea, and coffee.

    Science.gov (United States)

    Akagawa, Mitsugu; Shigemitsu, Tomoko; Suyama, Kyozo

    2005-10-01

    Oxidative deamination by various polyphenolic compounds is presumed to be due to the oxidative conversion of polyphenols to the corresponding quinones through autoxidation. Here we examined the oxidative deamination by the polyphenol-rich beverages green tea, black tea, and coffee at a physiological pH and temperature. Green tea, black tea, and coffee extracts oxidatively deaminated benzylamine and the lysine residues of bovine serum albumin to benzaldehyde and alpha-aminoadipic delta-semialdehyde residues, respectively, in sodium phosphate buffer (pH 7.4) at 37 degrees C in both the presence and absence of Cu2+, indicating the occurrence of an amine (lysyl) oxidase-like reaction. We also examined the effects of pH and metal ions on the reaction. The possible biological effects of drinking polyphenol-rich beverages on human are also discussed.

  13. Acetylation and glycation of fibrinogen in vitro occur at specific lysine residues in a concentration dependent manner: A mass spectrometric and isotope labeling study

    Energy Technology Data Exchange (ETDEWEB)

    Svensson, Jan, E-mail: jan.svensson@ki.se [Department of Molecular Medicine and Surgery, Karolinska Institutet, Karolinska University Hospital (Solna), SE-171 76 Stockholm (Sweden); Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, SE-182 88 Stockholm (Sweden); Bergman, Ann-Charlotte [Department of Molecular Medicine and Surgery, Karolinska Institutet, Karolinska University Hospital (Solna), SE-171 76 Stockholm (Sweden); Adamson, Ulf [Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, SE-182 88 Stockholm (Sweden); Blombaeck, Margareta [Department of Molecular Medicine and Surgery, Karolinska Institutet, Karolinska University Hospital (Solna), SE-171 76 Stockholm (Sweden); Wallen, Hakan; Joerneskog, Gun [Karolinska Institutet, Department of Clinical Sciences, Danderyd Hospital, SE-182 88 Stockholm (Sweden)

    2012-05-04

    Highlights: Black-Right-Pointing-Pointer Fibrinogen was incubated in vitro with glucose or aspirin. Black-Right-Pointing-Pointer Acetylations and glycations were found at twelve lysine sites by mass spectrometry. Black-Right-Pointing-Pointer The labeling by aspirin and glucose occurred dose-dependently. Black-Right-Pointing-Pointer No competition between glucose and aspirin for binding to fibrinogen was found. -- Abstract: Aspirin may exert part of its antithrombotic effects through platelet-independent mechanisms. Diabetes is a condition in which the beneficial effects of aspirin are less prominent or absent - a phenomenon called 'aspirin resistance'. We investigated whether acetylation and glycation occur at specific sites in fibrinogen and if competition between glucose and aspirin in binding to fibrinogen occurs. Our hypothesis was that such competition might be one explanation to 'aspirin resistance' in diabetes. After incubation of fibrinogen in vitro with aspirin (0.8 mM, 24 h) or glucose (100 mM, 5-10 days), we found 12 modified sites with mass spectrometric techniques. Acetylations in the {alpha}-chain: {alpha}K191, {alpha}K208, {alpha}K224, {alpha}K429, {alpha}K457, {alpha}K539, {alpha}K562, in the {beta}-chain: {beta}K233, and in the {gamma}-chain: {gamma}K170 and {gamma}K273. Glycations were found at {beta}K133 and {gamma}K75, alternatively {gamma}K85. Notably, the lysine 539 is a site involved in FXIII-mediated cross-linking of fibrin. With isotope labeling in vitro, using [{sup 14}C-acetyl]salicylic acid and [{sup 14}C]glucose, a labeling of 0.013-0.084 and 0.12-0.5 mol of acetylated and glycated adduct/mol fibrinogen, respectively, was found for clinically (12.9-100 {mu}M aspirin) and physiologically (2-8 mM glucose) relevant plasma concentrations. No competition between acetylation and glycation could be demonstrated. Thus, fibrinogen is acetylated at several lysine residues, some of which are involved in the cross-linking of

  14. Properties of Direct Coal Liquefaction Residue Modified Asphalt Mixture

    Directory of Open Access Journals (Sweden)

    Jie Ji

    2017-01-01

    Full Text Available The objectives of this paper are to use Direct Coal Liquefaction Residue (DLCR to modify the asphalt binders and mixtures and to evaluate the performance of modified asphalt mixtures. The dynamic modulus and phase angle of DCLR and DCLR-composite modified asphalt mixture were analyzed, and the viscoelastic properties of these modified asphalt mixtures were compared to the base asphalt binder SK-90 and Styrene-Butadiene-Styrene (SBS modified asphalt mixtures. The master curves of the asphalt mixtures were shown, and dynamic and viscoelastic behaviors of asphalt mixtures were described using the Christensen-Anderson-Marasteanu (CAM model. The test results show that the dynamic moduli of DCLR and DCLR-composite asphalt mixtures are higher than those of the SK-90 and SBS modified asphalt mixtures. Based on the viscoelastic parameters of CAM models of the asphalt mixtures, the high- and low-temperature performance of DLCR and DCLR-composite modified asphalt mixtures are obviously better than the SK-90 and SBS modified asphalt mixtures. In addition, the DCLR and DCLR-composite modified asphalt mixtures are more insensitive to the frequency compared to SK-90 and SBS modified asphalt mixtures.

  15. THE INDUSTRIAL UTILIZATION OF CHEMICAL MODIFIED AGRICULTURAL RESIDUES

    Institute of Scientific and Technical Information of China (English)

    Feng Xu; Runcang Sun; Huaiyu Zhan

    2004-01-01

    Various lignocellulosic materials such as wood,agricultural and forest residues has the potential to be valuable substitute for, or complement to,commercial sorbents for removing heavy metal ions or dyes from waste water or spilled oil from inland water or sea. More than 9 million tons of straw pulp are produced annually in china, which account for about 90% of the world′s total straw pulp. However,huge quantity of remain straw is not used as industrial raw material and is burnt in the fields or on the side of road. These resources can be chemical modified such as acetylation. Modified straws have the characteristics of low cost, high capacity, quick uptake, and easy to desorb. This paper reviews the current status of the technology for modified agricultural residues, which focus on hemicellulose and cellulose. The potential of these natural sorbents in main industry is also indicated.

  16. THE INDUSTRIAL UTILIZATION OF CHEMICAL MODIFIED AGRICULTURAL RESIDUES

    Institute of Scientific and Technical Information of China (English)

    FengXu; RuncangSun; HuaiyuZhan

    2004-01-01

    Various lignocellulosic materials such as wood, agricultural and forest residues has the potential to be valuable substitute for, or complement to, commercial sorbents for removing heavy metal ions or dyes from waste water or spilled oil from inland water or sea. More than 9 million tons of straw pulp are produced annually in china, which account for about 90% of the world's total straw pulp. However, huge quantity of remain straw is not used as industrial raw material and is burnt in the fields or on the side of road. These resources can be chemical modified such as acetylation. Modified straws have the characteristics of low cost, high capacity, quick uptake, and easy to desorb. This paper reviews the current status of the technology for modified agricultural residues, which focus on hemicellulose and cellulose. The potential of these natural sorbents in main industry is also indicated.

  17. Simultaneous Detection of Dopamine and Uric Acid Using a Poly(l-lysine/Graphene Oxide Modified Electrode

    Directory of Open Access Journals (Sweden)

    Yuehua Zhang

    2016-09-01

    Full Text Available A novel, simple and selective electrochemical method was investigated for the simultaneous detection of dopamine (DA and uric acid (UA on a poly(l-lysine/graphene oxide (GO modified glassy carbon electrode (PLL/GO/GCE by differential pulse voltammetry (DPV. The electrochemically prepared PLL/GO sensory platform toward the oxidation of UA and DA exhibited several advantages, including high effective surface area, more active sites and enhanced electrochemical activity. Compared to the PLL-modified GCE (PLL/GCE, GO-modified GCE and bare GCE, the PLL/GO/GCE exhibited an increase in the anodic potential difference and a remarkable enhancement in the current responses for both UA and DA. For the simultaneous detection of DA and UA, the detection limits of 0.021 and 0.074 μM were obtained, while 0.031 and 0.018 μM were obtained as the detection limits for the selective detection of UA and DA, using DPV in the linear concentration ranges of 0.5 to 20.0 and 0.5 to 35 μM, respectively. In addition, the PLL/GO/GCE demonstrated good reproducibility, long-term stability, excellent selectivity and negligible interference of ascorbic acid (AA. The proposed modified electrode was successfully implemented in the simultaneous detection of DA and UA in human blood serum, urine and dopamine hydrochloride injection with satisfactory results.

  18. [Preparation of OMC-Au/L-Lysine/Au modified glassy carbon electrode and the study on its detection response to hydroquinone and catechol].

    Science.gov (United States)

    Zhou, Yao-Yu; Tang, Lin; Li, Zhen; Liu, Yuan-Yuan; Yang, Gui-De; Wu, Meng-Shi; Lei, Xiao-Xia; Zheng, Guang-Ming

    2013-03-01

    Ordered mesoporous carbon-Au nanoparticles (OMC-Au) nanocomposites were synthesized by a one-step chemical reduction route, and an OMC-Au/L-Lysine/Au composite film-modified glassy carbon electrode (GCE) was constructed. The microstructure of OMC and OMC-Au/L-Lysine/Au composite films were characterized by SEM, and the preparation process of OMC-Au/L-Lysine/Au modified glassy carbon electrode was investigated using cyclic voltammetry and electrochemical impedance spectroscopy. The electrocatalytic oxidation of hydroquinone and catechol on the modified electrode was discussed by differential pulse voltammetry in this study, and a sensor for separate determination of hydroquinone and catechol based on OMC-Au/L-Lysine/Au modified glassy carbon electrode was developed. Under the optimal conditions, the cathodic peak current was linearly related to hydroquinone concentration over ranges from 1.0 x 10(-6) mol x L(-1) to 8.0 x 10(-4) mol x L(-1) with a detection limit of 3.0 x 10(-7) mol x L(-1), and linearly related to catechol concentration from 1.0 x 10(-7) mol x L(-1) to 8.0 x 10(-5) mol x L(-1) with a detection limit of 8.0 x 10(-7) mol x L(-1).

  19. Piscidin-1-analogs with double L- and D-lysine residues exhibited different conformations in lipopolysaccharide but comparable anti-endotoxin activities

    Science.gov (United States)

    Kumar, Amit; Mahajan, Mukesh; Awasthi, Bhanupriya; Tandon, Anshika; Harioudh, Munesh Kumar; Shree, Sonal; Singh, Pratiksha; Shukla, Praveen Kumar; Ramachandran, Ravishankar; Mitra, Kalyan; Bhattacharjya, Surajit; Ghosh, Jimut Kanti

    2017-01-01

    To become clinically effective, antimicrobial peptides (AMPs) should be non-cytotoxic to host cells. Piscidins are a group of fish-derived AMPs with potent antimicrobial and antiendotoxin activities but limited by extreme cytotoxicity. We conjectured that introduction of cationic residue(s) at the interface of polar and non-polar faces of piscidins may control their insertion into hydrophobic mammalian cell membrane and thereby reducing cytotoxicity. We have designed several novel analogs of piscidin-1 by substituting threonine residue(s) with L and D-lysine residue(s). L/D-lysine-substituted analogs showed significantly reduced cytotoxicity but exhibited either higher or comparable antibacterial activity akin to piscidin-1. Piscidin-1-analogs demonstrated higher efficacy than piscidin-1 in inhibiting lipopolysaccharide (LPS)-induced pro-inflammatory responses in THP-1 cells. T15,21K-piscidin-1 (0.5 mg/Kg) and T15,21dK-piscidin-1 (1.0 mg/Kg) demonstrated 100% survival of LPS (12.0 mg/Kg)-administered mice. High resolution NMR studies revealed that both piscidin-1 and T15,21K-piscidin-1 adopted helical structures, with latter showing a shorter helix, higher amphipathicity and cationic residues placed at optimal distances to form ionic/hydrogen bond with lipid A of LPS. Remarkably, T15,21dK-piscidin-1 showed a helix-loop-helix structure in LPS and its interactions with LPS could be sustained by the distance of separation of side chains of R7 and D-Lys-15 which is close to the inter-phosphate distance of lipid A. PMID:28051162

  20. Site-specific methylation and acetylation of lysine residues in the C-terminal domain (CTD) of RNA polymerase II

    Science.gov (United States)

    Voss, Kirsten; Forné, Ignasi; Descostes, Nicolas; Hintermair, Corinna; Schüller, Roland; Maqbool, Muhammad Ahmad; Heidemann, Martin; Flatley, Andrew; Imhof, Axel; Gut, Marta; Gut, Ivo; Kremmer, Elisabeth; Andrau, Jean-Christophe; Eick, Dirk

    2015-01-01

    Dynamic modification of heptad-repeats with the consensus sequence Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7 of RNA polymerase II (RNAPII) C-terminal domain (CTD) regulates transcription-coupled processes. Mass spectrometry analysis revealed that K7-residues in non-consensus repeats of human RNAPII are modified by acetylation, or mono-, di-, and tri-methylation. K7ac, K7me2, and K7me3 were found exclusively associated with phosphorylated CTD peptides, while K7me1 occurred also in non-phosphorylated CTD. The monoclonal antibody 1F5 recognizes K7me1/2 residues in CTD and reacts with RNAPIIA. Treatment of cellular extracts with phosphatase or of cells with the kinase inhibitor flavopiridol unmasked the K7me1/2 epitope in RNAPII0, consistent with the association of K7me1/2 marks with phosphorylated CTD peptides. Genome-wide profiling revealed high levels of K7me1/2 marks at the transcriptional start site of genes for sense and antisense transcribing RNAPII. The new K7 modifications further expand the mammalian CTD code to allow regulation of differential gene expression. PMID:26566685

  1. Site-specific methylation and acetylation of lysine residues in the C-terminal domain (CTD) of RNA polymerase II.

    Science.gov (United States)

    Voss, Kirsten; Forné, Ignasi; Descostes, Nicolas; Hintermair, Corinna; Schüller, Roland; Maqbool, Muhammad Ahmad; Heidemann, Martin; Flatley, Andrew; Imhof, Axel; Gut, Marta; Gut, Ivo; Kremmer, Elisabeth; Andrau, Jean-Christophe; Eick, Dirk

    2015-01-01

    Dynamic modification of heptad-repeats with the consensus sequence Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7 of RNA polymerase II (RNAPII) C-terminal domain (CTD) regulates transcription-coupled processes. Mass spectrometry analysis revealed that K7-residues in non-consensus repeats of human RNAPII are modified by acetylation, or mono-, di-, and tri-methylation. K7ac, K7me2, and K7me3 were found exclusively associated with phosphorylated CTD peptides, while K7me1 occurred also in non-phosphorylated CTD. The monoclonal antibody 1F5 recognizes K7me1/2 residues in CTD and reacts with RNAPIIA. Treatment of cellular extracts with phosphatase or of cells with the kinase inhibitor flavopiridol unmasked the K7me1/2 epitope in RNAPII0, consistent with the association of K7me1/2 marks with phosphorylated CTD peptides. Genome-wide profiling revealed high levels of K7me1/2 marks at the transcriptional start site of genes for sense and antisense transcribing RNAPII. The new K7 modifications further expand the mammalian CTD code to allow regulation of differential gene expression.

  2. Role of lysine and tryptophan residues in the biological activity of toxin VII (Ts gamma) from the scorpion Tityus serrulatus.

    Science.gov (United States)

    Hassani, O; Mansuelle, P; Cestèle, S; Bourdeaux, M; Rochat, H; Sampieri, F

    1999-02-01

    Toxin VII (TsVII), also known as Ts gamma, is the most potent neurotoxin in the venom of the Brazilian scorpion Tityus serrulatus. It has been purified to homogeneity using a new fast and efficient method. Chemical modification of TsVII with the tryptophan-specific reagent o-nitrophenylsulfenyl chloride yielded three modified derivatives (residues Trp39, Trp50 and Trp54). Acetylation of TsVII mostly generated the monoacetylated Lys12 derivative. No side reactions were detected, as indicated by endoproteinase Lys-C peptide mapping, Edman degradation and electrospray mass spectrometry. Circular dichroism and fluorimetric measurements showed that none of the chemical modifications altered the overall structure of the derivatives. The acetylation of Lys12 or the sulfenylation of Trp39 or Trp54 led to a loss of both toxicity in mice and apparent binding affinity for rat brain and cockroach synaptosomal preparations. Sulfenylation of Trp50, however, moderately affected the toxicity of TsVII in mice and had almost no effect on its binding properties. A 3-dimensional model of TsVII was constructed by homology modeling. It suggests that the most reactive residues (Lys12 and Trp39 and Trp54) are all important in the functional disruption of neuronal sodium channels by TsVII, and are close to each other in the hydrophobic conserved region.

  3. The roles of selected arginine and lysine residues of TAFI (Pro-CPU) in its activation to TAFIa by the thrombin-thrombomodulin complex.

    Science.gov (United States)

    Wu, Chengliang; Kim, Paul Y; Manuel, Reg; Seto, Marian; Whitlow, Marc; Nagashima, Mariko; Morser, John; Gils, Ann; Declerck, Paul; Nesheim, Michael E

    2009-03-13

    Thrombomodulin (TM) increases the catalytic efficiency of thrombin (IIa)-mediated activation of thrombin-activable fibrinolysis inhibitor (TAFI) 1250-fold. Negatively charged residues of the C-loop of TM-EGF-like domain 3 are required for TAFI activation. Molecular models suggested several positively charged residues of TAFI with which the C-loop residues could interact. Seven TAFI mutants were constructed to determine if these residues are required for efficient TAFI activation. TAFI wild-type or mutants were activated in the presence or absence of TM and the kinetic parameters of TAFI activation were determined. When the three consecutive lysine residues in the activation peptide of TAFI were substituted with alanine (K42/43/44A), the catalytic efficiencies for TAFI activation with TM decreased 8-fold. When other positively charged surface residues of TAFI (Lys-133, Lys-211, Lys-212, Arg-220, Lys-240, or Arg-275) were mutated to alanine, the catalytic efficiencies for TAFI activation with TM decreased by 1.7-2.7-fold. All decreases were highly statistically significant. In the absence of TM, catalytic efficiencies ranged from 2.8-fold lower to 1.24-fold higher than wild-type. None of these, except the 2.8-fold lower value, was statistically significant. The average half-life of the TAFIa mutants was 8.1+/-0.6 min, and that of wild type was 8.4+/-0.3 min at 37 degrees C. Our data show that these residues are important in the activation of TAFI by IIa, especially in the presence of TM. Whether the mutated residues promote a TAFI-TM or TAFI-IIa interaction remains to be determined. In addition, these residues do not influence spontaneous inactivation of TAFIa.

  4. Expansion of the Lysine Acylation Landscape

    DEFF Research Database (Denmark)

    Olsen, Christian A.

    2012-01-01

    Leaving marks: The number of known posttranslational modifications for lysine has been expanded considerably. In addition to acetylation of side-chain amino functionalities of lysine residues in proteins, crotonylation, succinylation, and malonylation have now been identified as posttranslational...

  5. EFFECTS OF SUPPLEMENTAL LYSINE ON PERFORMANCE, ANTIBODY TITER AND RECTAL TEMPERATURE IN RESPONSE TO A MODIFIED-LIVE VIRAL VACCINE IN NEONATAL CALVES

    Directory of Open Access Journals (Sweden)

    Kate Sharon

    2014-01-01

    Full Text Available Infectious Bovine Rhinotracheitis (IBR, caused by bovine herpesvirus-1, contributes significantly to economic losses in the dairy and beef cattle industry. Lysine has been shown reduce virulence of herpesviruses in felids and humans. Our objective was to evaluate the effects of supplemental lysine on serum IBR antibody titer and rectal temperature in response to a modified-live Intranasal (IN or Intramuscular (IM respiratory-virus vaccination. Sixty-four neonatal Holstein bull calves (7±2 d of age; BW = 37±4.2 kg were used in a completely randomized design. Calves were fed milk replacer supplemented with either 17 g/d L-lysine monohydrochloride (LYS; 28 calves or an equivalent amount of casein (CAS; 28 calves for 42 d. Calves were then vaccinated with either an IN IBR-Parainfluenza virus-3 (PI3 or an IM (IBR-PI3-bovine viral diarrhea type I and II, bovine respiratory syncytial virus modified-live vaccine on d 36. A control group (8 calves received no supplement or vaccination. All calves were housed in individual calf pens (1.2×2.1 m. Daily feed intakes were monitored and BW measured weekly. Calves were bled on d 0, 35, 36, 37 and 42. Temperature data loggers were attached to rectal probes and temperatures were recorded every 5 min from d 28 to d 42. No significant differences were determined for average performance, rectal temperature, or IBR antibody titers with either IN or IM vaccinations between LYS and CAS treated calves (p>0.10. However, serum urea nitrogen and the ratio of serum lysine: Arginine increased (p<0.05 for LYS compared to CAS calves. These results suggest that supplementing lysine impacts nitrogen metabolism but does not alter the response to IBR vaccination or animal performance in neonatal Holstein calves.

  6. Thermo-sensitive liposomes loaded with doxorubicin and lysine modified single-walled carbon nanotubes as tumor-targeting drug delivery system.

    Science.gov (United States)

    Zhu, Xiali; Xie, Yingxia; Zhang, Yingjie; Huang, Heqing; Huang, Shengnan; Hou, Lin; Zhang, Huijuan; Li, Zhi; Shi, Jinjin; Zhang, Zhenzhong

    2014-11-01

    This report focuses on the thermo-sensitive liposomes loaded with doxorubicin and lysine-modified single-walled carbon nanotube drug delivery system, which was designed to enhance the anti-tumor effect and reduce the side effects of doxorubicin. Doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes was prepared by reverse-phase evaporation method, the mean particle size was 232.0 ± 5.6 nm, and drug entrapment efficiency was 86.5 ± 3.7%. The drug release test showed that doxorubicin released more quickly at 42℃ than at 37℃. Compared with free doxorubicin, doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes could efficiently cross the cell membranes and afford higher anti-tumor efficacy on the human hepatic carcinoma cell line (SMMC-7721) cells in vitro. For in vivo experiments, the relative tumor volumes of the sarcomaia 180-bearing mice in thermo-sensitive liposomes group and doxorubicin group were significantly smaller than those of N.S. group. Meanwhile, the combination of near-infrared laser irradiation at 808 nm significantly enhanced the tumor growth inhibition both on SMMC-7721 cells and the sarcomaia 180-bearing mice. The quality of life such as body weight, mental state, food and water intake of sarcomaia 180 tumor-bearing mice treated with doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes were much higher than those treated with doxorubicin. In conclusion, doxorubicin-lysine/single-walled carbon nanotube-thermo-sensitive liposomes combined with near-infrared laser irradiation at 808 nm may potentially provide viable clinical strategies for targeting delivery of anti-cancer drugs.

  7. Poly-L-lysine-modified reduced graphene oxide stabilizes the copper nanoparticles with higher water-solubility and long-term additively antibacterial activity.

    Science.gov (United States)

    Ouyang, Yu; Cai, Xiang; Shi, QingShan; Liu, Lili; Wan, Dongliang; Tan, Shaozao; Ouyang, Yousheng

    2013-07-01

    In order to improve the water-solubility and long-term antibacterial activity of copper nanoparticles (CuNPs), a poly-L-lysine-modified reduced graphene oxide (PLL-rGO) was used as the carrier of CuNPs, and a poly-L-lysine/reduced graphene oxide/copper nanoparticles (PLL-rGO-CuNPs) hybrid was prepared by anchoring the CuNPs on the reduced graphene oxide surface. The novel PLL-rGO-CuNPs hybrid was characterized and the antibacterial activity of it on gram-negative Escherichia coli and Gram-positive Staphylococcus aureus was tested. Such a hybrid showed additively antibacterial activity, and the CuNPs on PLL-rGO were more stable than those on polyvinyl pyrrolidone, resulting in long-term additively antibacterial effect. Meanwhile, this hybrid showed excellent water-solubility, suggesting great potential application in microbial control.

  8. Lysine221 is the general base residue of the isochorismate synthase from Pseudomonas aeruginosa (PchA) in a reaction that is diffusion limited.

    Science.gov (United States)

    Meneely, Kathleen M; Luo, Qianyi; Dhar, Prajnaparamita; Lamb, Audrey L

    2013-10-01

    The isochorismate synthase from Pseudomonas aeruginosa (PchA) catalyzes the conversion of chorismate to isochorismate, which is subsequently converted by a second enzyme (PchB) to salicylate for incorporation into the salicylate-capped siderophore pyochelin. PchA is a member of the MST family of enzymes, which includes the structurally homologous isochorismate synthases from Escherichia coli (EntC and MenF) and salicylate synthases from Yersinia enterocolitica (Irp9) and Mycobacterium tuberculosis (MbtI). The latter enzymes generate isochorismate as an intermediate before generating salicylate and pyruvate. General acid-general base catalysis has been proposed for isochorismate synthesis in all five enzymes, but the residues required for the isomerization are a matter of debate, with both lysine221 and glutamate313 proposed as the general base (PchA numbering). This work includes a classical characterization of PchA with steady state kinetic analysis, solvent kinetic isotope effect analysis and by measuring the effect of viscosogens on catalysis. The results suggest that isochorismate production from chorismate by the MST enzymes is the result of general acid-general base catalysis with a lysine as the base and a glutamic acid as the acid, in reverse protonation states. Chemistry is determined to not be rate limiting, favoring the hypothesis of a conformational or binding step as the slow step.

  9. Lysine residues in N-terminal and C-terminal regions of human histone H2A are targets for biotinylation by biotinidase.

    Science.gov (United States)

    Chew, Yap Ching; Camporeale, Gabriela; Kothapalli, Nagarama; Sarath, Gautam; Zempleni, Janos

    2006-04-01

    In eukaryotic cell nuclei, DNA associates with the core histones H2A, H2B, H3 and H4 to form nucleosomal core particles. DNA binding to histones is regulated by posttranslational modifications of N-terminal tails (e.g., acetylation and methylation of histones). These modifications play important roles in the epigenetic control of chromatin structure. Recently, evidence that biotinidase and holocarboxylase synthetase (HCS) catalyze the covalent binding of biotin to histones has been provided. The primary aim of this study was to identify biotinylation sites in histone H2A and its variant H2AX. Secondary aims were to determine whether acetylation and methylation of histone H2A affect subsequent biotinylation and whether biotinidase and HCS localize to the nucleus in human cells. Biotinylation sites were identified using synthetic peptides as substrates for biotinidase. These studies provided evidence that K9 and K13 in the N-terminus of human histones H2A and H2AX are targets for biotinylation and that K125, K127 and K129 in the C-terminus of histone H2A are targets for biotinylation. Biotinylation of lysine residues was decreased by acetylation of adjacent lysines but was increased by dimethylation of adjacent arginines. The existence of biotinylated histone H2A in vivo was confirmed by using modification-specific antibodies. Antibodies to biotinidase and HCS localized primarily to the nuclear compartment, consistent with a role for these enzymes in regulating chromatin structure. Collectively, these studies have identified five novel biotinylation sites in human histones; histone H2A is unique among histones in that its biotinylation sites include amino acid residues from the C-terminus.

  10. Substitution of glutamate residue by lysine in the dimerization domain affects DNA binding ability of HapR by inducing structural deformity in the DNA binding domain.

    Directory of Open Access Journals (Sweden)

    Richa Singh

    Full Text Available HapR has been given the status of a high cell density master regulatory protein in Vibrio cholerae. Though many facts are known regarding its structural and functional aspects, much still can be learnt from natural variants of the wild type protein. This work aims at investigating the nature of functional inertness of a HapR natural variant harboring a substitution of a conserved glutamate residue at position 117 which participates in forming a salt bridge by lysine (HapRV2G-E(117K. Experimental evidence presented here reveals the inability of this variant to interact with various cognate promoters by in vitro gel shift assay. Furthermore, the elution profiles of HapRV2G-E(117K protein along with the wild type functional HapRV2G in size-exclusion chromatography as well as circular dichroism spectra did not reflect any significant differences in its structure, thereby indicating the intactness of dimer in the variant protein. To gain further insight into the global shape of the proteins, small angle X-ray scattering analysis (SAXS was performed. Intriguingly, increased radius of gyration of HapRV2G-E(117K of 27.5 Å in comparison to the wild type protein from SAXS data analyses implied a significant alteration in the global shape of the dimeric HapRV2G-E(117K protein. Structure reconstruction brought forth that the DNA binding domains were substantially "parted away" in this variant. Taken together, our data illustrates that substitution of the conserved glutamate residue by lysine in the dimerization domain induces separation of the two DNA binding domains from their native-like positioning without altering the dimeric status of HapR variant.

  11. Substitution of glutamate residue by lysine in the dimerization domain affects DNA binding ability of HapR by inducing structural deformity in the DNA binding domain.

    Science.gov (United States)

    Singh, Richa; Rathore, Yogendra Singh; Singh, Naorem Santa; Peddada, Nagesh; Ashish; Raychaudhuri, Saumya

    2013-01-01

    HapR has been given the status of a high cell density master regulatory protein in Vibrio cholerae. Though many facts are known regarding its structural and functional aspects, much still can be learnt from natural variants of the wild type protein. This work aims at investigating the nature of functional inertness of a HapR natural variant harboring a substitution of a conserved glutamate residue at position 117 which participates in forming a salt bridge by lysine (HapRV2G-E(117)K). Experimental evidence presented here reveals the inability of this variant to interact with various cognate promoters by in vitro gel shift assay. Furthermore, the elution profiles of HapRV2G-E(117)K protein along with the wild type functional HapRV2G in size-exclusion chromatography as well as circular dichroism spectra did not reflect any significant differences in its structure, thereby indicating the intactness of dimer in the variant protein. To gain further insight into the global shape of the proteins, small angle X-ray scattering analysis (SAXS) was performed. Intriguingly, increased radius of gyration of HapRV2G-E(117)K of 27.5 Å in comparison to the wild type protein from SAXS data analyses implied a significant alteration in the global shape of the dimeric HapRV2G-E(117)K protein. Structure reconstruction brought forth that the DNA binding domains were substantially "parted away" in this variant. Taken together, our data illustrates that substitution of the conserved glutamate residue by lysine in the dimerization domain induces separation of the two DNA binding domains from their native-like positioning without altering the dimeric status of HapR variant.

  12. Cationic liposomes enhance targeted delivery and expression of exogenous DNA mediated by N-terminal modified poly(L-lysine)-antibody conjugate in mouse lung endothelial cells.

    Science.gov (United States)

    Trubetskoy, V S; Torchilin, V P; Kennel, S; Huang, L

    1992-07-15

    A new and improved system for targeted gene delivery and expression is described. Transfection efficiency of N-terminal modified poly(L-lysine) (NPLL) conjugated with anti-thrombomodulin antibody 34A can be improved by adding to the system a lipophilic component, cationic liposomes. DNA, antibody conjugate and cationic liposomes form a ternary electrostatic complex which preserves the ability to bind specifically to the target cells. At the same time the addition of liposomes enhance the specific transfection efficiency of antibody-polylysine/DNA binary complex by 10 to 20-fold in mouse lung endothelial cells in culture.

  13. Use of N-terminal modified poly(L-lysine)-antibody conjugate as a carrier for targeted gene delivery in mouse lung endothelial cells.

    Science.gov (United States)

    Trubetskoy, V S; Torchilin, V P; Kennel, S J; Huang, L

    1992-01-01

    A DNA targeted delivery and expression system has been designed based on an N-terminal modified poly(L-lysine) (NPLL)-antibody conjugate, which readily forms a complex with plasmid DNA. Monoclonal antibodies against the cell-surface thrombomodulin conjugated with NPLL were used for targeted delivery of foreign plasmid DNA to an antigen-expressing mouse lung endothelial cell line in vitro and to mouse lungs in vivo. In both cases significant amounts of DNA can be specifically bound to the target cells or tissues. Specific gene expression was observed in the treated mouse lung endothelial cells.

  14. Contribution of a lysine residue in the first transmembrane segment to the selectivity filter region in the CFTR chloride channel.

    Science.gov (United States)

    Negoda, Alexander; El Hiani, Yassine; Cowley, Elizabeth A; Linsdell, Paul

    2017-02-21

    The anion selectivity and conductance of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel are determined predominantly by interactions between permeant anions and the narrow region of the channel pore. This narrow region has therefore been described as functioning as the "selectivity filter" of the channel. Multiple pore-lining transmembrane segments (TMs) have previously been shown to contribute to the selectivity filter region. However, little is known about the three-dimensional organization of this region, or how multiple TMs combine to determine its functional properties. In the present study we have used patch clamp recording to identify changes in channel function associated with the formation of disulfide cross-links between cysteine residues introduced into different TMs within the selectivity filter. Cysteine introduced at position L102 in TM1 was able to form disulfide bonds with F337C and T338C in TM6, two positions that are known to play key roles in determining anion permeation properties. Consistent with this proximal arrangement of L102, F337 and T338, different mutations at L102 altered anion selectivity and conductance properties in a way that suggests that this residue plays an important role in determining selectivity filter function, albeit a much lesser role than that of F337. These results suggest an asymmetric three-dimensional arrangement of the key selectivity filter region of the pore, as well as having important implications regarding the molecular mechanism of anion permeation.

  15. Lysine methylation: beyond histones

    Institute of Scientific and Technical Information of China (English)

    Xi Zhang; Hong Wen; Xiaobing Shi

    2012-01-01

    Posttranslational modifications (PTMs) of histone proteins,such as acetylation,methylation,phosphorylation,and ubiquitylation,play essential roles in regulating chromatin dynamics.Combinations of different modifications on the histone proteins,termed 'histone code' in many cases,extend the information potential of the genetic code by regulating DNA at the epigenetic level.Many PTMs occur on non-histone proteins as well as histones,regulating protein-protein interactions,stability,localization,and/or enzymatic activities of proteins involved in diverse cellular processes.Although protein phosphorylation,ubiquitylation,and acetylation have been extensively studied,only a few proteins other than histones have been reported that can be modified by lysine methylation.This review summarizes the current progress on lysine methylation of nonhistone proteins,and we propose that lysine methylation,like phosphorylation and acetylation,is a common PTM that regulates proteins in diverse cellular processes.

  16. SIRT1 Regulates Thyroid-Stimulating Hormone Release by Enhancing PIP5Kgamma Activity through Deacetylation of Specific Lysine Residues in Mammals.

    Directory of Open Access Journals (Sweden)

    Sayaka Akieda-Asai

    Full Text Available BACKGROUND: SIRT1, a NAD-dependent deacetylase, has diverse roles in a variety of organs such as regulation of endocrine function and metabolism. However, it remains to be addressed how it regulates hormone release there. METHODOLOGY/PRINCIPAL FINDINGS: Here, we report that SIRT1 is abundantly expressed in pituitary thyrotropes and regulates thyroid hormone secretion. Manipulation of SIRT1 level revealed that SIRT1 positively regulated the exocytosis of TSH-containing granules. Using LC/MS-based interactomics, phosphatidylinositol-4-phosphate 5-kinase (PIP5Kgamma was identified as a SIRT1 binding partner and deacetylation substrate. SIRT1 deacetylated two specific lysine residues (K265/K268 in PIP5Kgamma and enhanced PIP5Kgamma enzyme activity. SIRT1-mediated TSH secretion was abolished by PIP5Kgamma knockdown. SIRT1 knockdown decreased the levels of deacetylated PIP5Kgamma, PI(4,5P(2, and reduced the secretion of TSH from pituitary cells. These results were also observed in SIRT1-knockout mice. CONCLUSIONS/SIGNIFICANCE: Our findings indicated that the control of TSH release by the SIRT1-PIP5Kgamma pathway is important for regulating the metabolism of the whole body.

  17. Conserved aspartate and lysine residues of RcsB are required for amylovoran biosynthesis, virulence, and DNA binding in Erwinia amylovora.

    Science.gov (United States)

    Ancona, Veronica; Chatnaparat, Tiyakhon; Zhao, Youfu

    2015-08-01

    In Erwinia amylovora, the Rcs phosphorelay system is essential for amylovoran production and virulence. To further understand the role of conserved aspartate residue (D56) in the phosphor receiver (PR) domain and lysine (K180) residue in the function domain of RcsB, amino acid substitutions of RcsB mutant alleles were generated by site-directed mutagenesis and complementation of various rcs mutants were performed. A D56E substitution of RcsB, which mimics the phosphorylation state of RcsB, complemented the rcsB mutant, resulting in increased amylovoran production and gene expression, reduced swarming motility, and restored pathogenicity. In contrast, D56N and K180A or K180Q substitutions of RcsB did not complement the rcsB mutant. Electrophoresis mobility shift assays showed that D56E, but not D56N, K180Q and K180A substitutions of RcsB bound to promoters of amsG and flhD, indicating that both D56 and K180 are required for DNA binding. Interestingly, the RcsBD56E allele could also complement rcsAB, rcsBC and rcsABCD mutants with restored virulence and increased amylovoran production, indicating that RcsB phosphorylation is essential for virulence of E. amylovora. In addition, mutations of T904 and A905, but not phosphorylation mimic mutation of D876 in the PR domain of RcsC, constitutively activate the Rcs system, suggesting that phosphor transfer is required for activating the Rcs system and indicating both A905 and T904 are required for the phosphatase activity of RcsC. Our results demonstrated that RcsB phosphorylation and dephosphorylation, phosphor transfer from RcsC are essential for the function of the Rcs system, and also suggested that constitutive activation of the Rcs system could reduce the fitness of E. amylovora.

  18. Unintended Changes in Genetically Modified Rice Expressing the Lysine-Rich Fusion Protein Gene Revealed by a Proteomics Approach

    Institute of Scientific and Technical Information of China (English)

    ZHAO Xiang-xiang; TANG Tang; LIU Fu-xia; LU Chang-li; HU Xiao-lan; JI Li-lian; LIU Qiao-quan

    2013-01-01

    Development of new technologies for evaluating genetically modiifed (GM) crops has revealed that there are unintended insertions and expression changes in GM crops. Proifling techniques are non-targeted approaches and are capable of detecting more unintended changes in GM crops. Here, we report the application of a comparative proteomic approach to investigate the protein proifle differences between a GM rice line, which has a lysine-rich protein gene, and its non-transgenic parental line. Proteome analysis by two-dimensional gel electrophoresis (2-DE) and mass spectrum analysis of the seeds identiifed 22 differentially expressed protein spots. Apart from a number of glutelins that were detected as targeted proteins in the GM line, the majority of the other changed proteins were involved in carbohydrate metabolism, protein synthesis and stress responses. These results indicated that the altered proteins were not associated with plant allergens or toxicity.

  19. Regeneration of poly-L-lysine modified carbon electrodes in the accumulation and cathodic stripping voltammetric determination of the cromoglycate anion.

    Science.gov (United States)

    Pereira, F C; Fogg, A G; Zanoni, M V B

    2003-07-27

    Cromoglycate is accumulated on a poly-L-lysine (PLL) modified carbon electrode best from pH 4 solution, where it is anionic and the PLL is cationic, and at which pH the cromoglycate gives a good reduction peak at -0.82 V. The PLL film can be regenerated readily by washing the electrode with 3 M sodium hydroxide solution, in which the PLL is deprotonated. Regeneration of the film is not required as frequently when larger amounts of PLL are incorporated into it. This allows standard addition procedures to be carried out without regenerating the electrode. Linear calibration graphs have been obtained typically in the range 0.1-1.5 microg ml(-1). Detection limits have been calculated to be 10 ng ml(-1). The standard addition method has been applied satisfactorily to diluted urine solutions.

  20. The valine and lysine residues in the conserved FxVTxK motif are important for the function of phylogenetically distant plant cellulose synthases.

    Science.gov (United States)

    Slabaugh, Erin; Scavuzzo-Duggan, Tess; Chaves, Arielle; Wilson, Liza; Wilson, Carmen; Davis, Jonathan K; Cosgrove, Daniel J; Anderson, Charles T; Roberts, Alison W; Haigler, Candace H

    2016-05-01

    Cellulose synthases (CESAs) synthesize the β-1,4-glucan chains that coalesce to form cellulose microfibrils in plant cell walls. In addition to a large cytosolic (catalytic) domain, CESAs have eight predicted transmembrane helices (TMHs). However, analogous to the structure of BcsA, a bacterial CESA, predicted TMH5 in CESA may instead be an interfacial helix. This would place the conserved FxVTxK motif in the plant cell cytosol where it could function as a substrate-gating loop as occurs in BcsA. To define the functional importance of the CESA region containing FxVTxK, we tested five parallel mutations in Arabidopsis thaliana CESA1 and Physcomitrella patens CESA5 in complementation assays of the relevant cesa mutants. In both organisms, the substitution of the valine or lysine residues in FxVTxK severely affected CESA function. In Arabidopsis roots, both changes were correlated with lower cellulose anisotropy, as revealed by Pontamine Fast Scarlet. Analysis of hypocotyl inner cell wall layers by atomic force microscopy showed that two altered versions of Atcesa1 could rescue cell wall phenotypes observed in the mutant background line. Overall, the data show that the FxVTxK motif is functionally important in two phylogenetically distant plant CESAs. The results show that Physcomitrella provides an efficient model for assessing the effects of engineered CESA mutations affecting primary cell wall synthesis and that diverse testing systems can lead to nuanced insights into CESA structure-function relationships. Although CESA membrane topology needs to be experimentally determined, the results support the possibility that the FxVTxK region functions similarly in CESA and BcsA.

  1. Host determinant residue lysine 627 lies on the surface of a discrete, folded domain of influenza virus polymerase PB2 subunit.

    Directory of Open Access Journals (Sweden)

    Franck Tarendeau

    Full Text Available Understanding how avian influenza viruses adapt to human hosts is critical for the monitoring and prevention of future pandemics. Host specificity is determined by multiple sites in different viral proteins, and mutation of only a limited number of these sites can lead to inter-species transmission. Several of these sites have been identified in the viral polymerase, the best characterised being position 627 in the PB2 subunit. Efficient viral replication at the relatively low temperature of the human respiratory tract requires lysine 627 rather than the glutamic acid variant found systematically in avian viruses. However, the molecular mechanism by which any of these host specific sites determine host range are unknown, although adaptation to host factors is frequently evoked. We used ESPRIT, a library screening method, to identify a new PB2 domain that contains a high density of putative host specific sites, including residue 627. The X-ray structure of this domain (denoted the 627-domain exhibits a novel fold with the side-chain of Lys627 solvent exposed. The structure of the K627E mutated domain shows no structural differences but the charge reversal disrupts a striking basic patch on the domain surface. Five other recently proposed host determining sites of PB2 are also located on the 627-domain surface. The structure of the complete C-terminal region of PB2 comprising the 627-domain and the previously identified NLS-domain, which binds the host nuclear import factor importin alpha, was also determined. The two domains are found to pack together with a largely hydrophilic interface. These data enable a three-dimensional mapping of approximately half of PB2 sites implicated in cross-species transfer onto a single structural unit. Their surface location is consistent with roles in interactions with other viral proteins or host factors. The identification and structural characterization of these well-defined PB2 domains will help design

  2. Quaternary complexes modified from pDNA and poly-l-lysine complexes to enhance pH-buffering effect and suppress cytotoxicity.

    Science.gov (United States)

    Kodama, Yukinobu; Yatsugi, Yuiko; Kitahara, Takashi; Kurosaki, Tomoaki; Egashira, Kanoko; Nakashima, Mikiro; Muro, Takahiro; Nakagawa, Hiroo; Higuchi, Norihide; Nakamura, Tadahiro; Sasaki, Hitoshi

    2015-04-01

    We developed a modified complex of pDNA and poly-l-lysine (PLL) by the addition of poly-l-histidine (PLH) and γ-polyglutamic acid (γ-PGA) to enhance its pH-buffering effect and suppress cytotoxicity. The binary and ternary complexes of pDNA with PLL or/and PLH showed particle sizes of approximately 52-76 nm with cationic surface charge. The ternary complexes showed much higher gene expression than the binary complexes with PLL. The mixed solution of PLL and PLH showed higher buffering capacity than PLL solution. The high gene expression of ternary complexes was reduced by bafilomycin A1 . These results indicated the addition of PLH to PLL complexes promoted endosomal escape by enhancing the pH-buffering effect. The binary and ternary complexes showed cytotoxicity and blood agglutination because of their cationic surface charge. We therefore developed quaternary complexes by the addition of anionic γ-PGA, which was reported to decrease the toxicity of cationic complexes. In fact, quaternary complexes showed no cytotoxicity and blood agglutination. Also, quaternary complexes showed higher gene expression than ternary complexes regardless of their anionic surface charge. Quaternary complexes showed selectively high gene expression in the spleen after their intravenous administration. Thus, we successfully developed the quaternary complexes with high gene expression and no toxicity.

  3. Molecular and structural insight into lysine selection on substrate and ubiquitin lysine 48 by the ubiquitin-conjugating enzyme Cdc34

    DEFF Research Database (Denmark)

    Suryadinata, Randy; Holien, Jessica K; Yang, George

    2013-01-01

    The attachment of ubiquitin (Ub) to lysines on substrates or itself by ubiquitin-conjugating (E2) and ubiquitin ligase (E3) enzymes results in protein ubiquitination. Lysine selection is important for generating diverse substrate-Ub structures and targeting proteins to different fates; however......, the mechanisms of lysine selection are not clearly understood. The positioning of lysine(s) toward the E2/E3 active site and residues proximal to lysines are critical in their selection. We investigated determinants of lysine specificity of the ubiquitin-conjugating enzyme Cdc34, toward substrate and Ub lysines....... Evaluation of the relative importance of different residues positioned -2, -1, +1 and +2 toward ubiquitination of its substrate, Sic1, on lysine 50 showed that charged residues in the -1 and -2 positions negatively impact on ubiquitination. Modeling suggests that charged residues at these positions alter...

  4. An autoantibody against N{sup {epsilon}}-(carboxyethyl)lysine (CEL): Possible involvement in the removal of CEL-modified proteins by macrophages

    Energy Technology Data Exchange (ETDEWEB)

    Mera, Katsumi [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Nagai, Ryoji, E-mail: nagai-883@umin.ac.jp [Department of Food and Nutrition, Laboratory of Nutritional Science and Biochemistry, Japan Women' s University, Tokyo (Japan); Takeo, Kazuhiro; Izumi, Miyoko [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Maruyama, Toru [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Center for Clinical Pharmaceutical Science, Kumamoto University, Kumamoto (Japan); Otagiri, Masaki [Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto (Japan); Faculty of Pharmaceutical Sciences, Sojo University, Kumamoto (Japan)

    2011-04-08

    Highlights: {yields} A higher amount of autoantibody against CEL than that of other AGEs was observed in human plasma. {yields} The purified human anti-CEL autoantibody specifically reacted with CEL. {yields} Anti-CEL antibody accelerated the uptake of {sup 125}I-CEL-HSA by macrophage in vitro. {yields} Endocytic uptake of {sup 125}I-CEL-HSA by mice liver was accelerated in the presence of anti-CEL antibody. -- Abstract: Advanced glycation end products (AGEs) are believed to play a significant role in the development of diabetic complications. In this study, we measured the levels of autoantibodies against several AGE structures in healthy human plasma and investigated the physiological role of the autoantibodies. A high titer of the autoantibody against N{sup {epsilon}}-(carboxyethyl)lysine (CEL) was detected in human plasma compared with other AGE structures such as CML and pentosidine. The purified human anti-CEL autoantibody reacted with CEL-modified human serum albumin (CEL-HSA), but not CML-HSA. A rabbit polyclonal anti-CEL antibody, used as a model autoantibody against CEL, accelerated the uptake of CEL-HSA by macrophages, but did not enhance the uptake of native HSA. Furthermore, when {sup 125}I-labeled CEL-HSA was injected into the tail vein of mice, accumulation of {sup 125}I-CEL-HSA in the liver was accelerated by co-injection of the rabbit anti-CEL antibody. These results demonstrate that the autoantibody against CEL in plasma may play a role in the macrophage uptake of CEL-modified proteins.

  5. Effect of bacteriophage lysin on lysogens

    Institute of Scientific and Technical Information of China (English)

    Balaji Subramanyam; Vanaja Kumar

    2011-01-01

    Objective: To study the effect of phage lysin on the growth of lysogens. Methods: Sputum specimens processed by modified Petroff's method were respectively treated with phagebiotics in combination with lysin and lysin alone. The specimens were incubated at 37℃ for 4 days. At the end of day 1, 2, 3 and day 4, the specimens were streaked on blood agar plates and incubated at 37℃ for 18-24 hours. The growth of normal flora observed after day 1 was considered as lysogens.Results:When specimens treated with lysin alone, lysogen formation was avoided and normal flora was controlled. Conclusions: Lysin may have no effect on the growth of lysogens. Sputum specimens treated with phagebiotics-lysin showed the growth of lysogens.

  6. Nucleosome alterations caused by mutations at modifiable histone residues in Saccharomyces cerevisiae.

    Science.gov (United States)

    Liu, Hongde; Wang, Pingyan; Liu, Lingjie; Min, Zhu; Luo, Kun; Wan, Yakun

    2015-10-26

    Nucleosome organization exhibits dynamic properties depending on the cell state and environment. Histone proteins, fundamental components of nucleosomes, are subject to chemical modifications on particular residues. We examined the effect of substituting modifiable residues of four core histones with the non-modifiable residue alanine on nucleosome dynamics. We mapped the genome-wide nucleosomes in 22 histone mutants of Saccharomyces cerevisiae and compared the nucleosome alterations relative to the wild-type strain. Our results indicated that different types of histone mutation resulted in different phenotypes and a distinct reorganization of nucleosomes. Nucleosome occupancy was altered at telomeres, but not at centromeres. The first nucleosomes upstream (-1) and downstream (+1) of the transcription start site (TSS) were more dynamic than other nucleosomes. Mutations in histones affected the nucleosome array downstream of the TSS. Highly expressed genes, such as ribosome genes and genes involved in glycolysis, showed increased nucleosome occupancy in many types of histone mutant. In particular, the H3K56A mutant exhibited a high percentage of dynamic genomic regions, decreased nucleosome occupancy at telomeres, increased occupancy at the +1 and -1 nucleosomes, and a slow growth phenotype under stress conditions. Our findings provide insight into the influence of histone mutations on nucleosome dynamics.

  7. Oligo-lysine Induced Formation of Silica Particles in Neutral Silicate Solution

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Oligo-(lysine)n (n = 1-4) containing different numbers of lysine residues was used to induce the condensation of silicic acid to form silica particles in neutral silicate solution. It was found that the condensation rate and the formation of silica particles are dependent on the number of lysine residues in an oligo-lysine. Oligo-lysine with more lysine residues can link more silicic acid together to form a matrix that promotes the effective aggregation of the condensed silica pieces to form large silica particles.

  8. Hypochlorite-induced damage to proteins: formation of nitrogen-centred radicals from lysine residues and their role in protein fragmentation.

    Science.gov (United States)

    Hawkins, C L; Davies, M J

    1998-01-01

    Stimulated monocytes and neutrophils generate hypochlorite (HOCl) via the release of the enzyme myeloperoxidase and hydrogen peroxide. HOCl damages proteins by reaction with amino acid side-chains or backbone cleavage. Little information is available about the mechanisms and intermediates involved in these reactions. EPR spin trapping has been employed to identify radicals on proteins, peptides and amino acids after treatment with HOCl. Reaction with HOCl gives both high- and low-molecular-mass nitrogen-centred, protein-derived radicals; the yield of the latter increases with both higher HOCl:protein ratios and enzymic digestion. These radicals, which arise from lysine side-chain amino groups, react with ascorbate, glutathione and Trolox. Reaction of HOCl-treated proteins with excess methionine eliminates radical formation, which is consistent with lysine-derived chloramines (via homolysis of N-Cl bonds) being the radical source. Incubation of HOCl-treated proteins, after removal of excess oxidant, gives rise to both nitrogen-centred radicals, over a period of hours, and time-dependent fragmentation of the protein. Treatment with excess methionine or antioxidants (Trolox, ascorbate, glutathione) protects against fragmentation; urate and bilirubin do not. Chloramine formation and nitrogen-centred radicals are therefore key species in HOCl-induced protein fragmentation. PMID:9620862

  9. Modified Layer-Removal Method for Measurement of Residual Stress in Pre-stretched Aluminium Alloy Plate

    Institute of Scientific and Technical Information of China (English)

    Liangbao Liu; Jianfei Sun; Wuyi Chen; Pengfei Sun

    2015-01-01

    Residual stress is one of the factors affecting the machining deformation of monolithic structure parts in the aviation industry. Thus, the studies on machining deformation rules induced by residual stresses largely depend on correctly and efficiently measuring the residual stresses of workpieces. A modified layer⁃removal method is proposed to measure residual stress by analysing the characteristics of a traditional layer⁃removal method. The coefficients of strain release are then deduced according to the simulation results using the finite element method ( FEM) . Moreover, the residual stress in a 7075T651 aluminium alloy plate is measured using the proposed method, and the results are then analyzed and compared with the data obtained by the traditional methods. The analysis indicates that the modified layer⁃removal method is effective and practical for measuring the residual stress distribution in pre⁃stretched aluminium alloy plates.

  10. Study of enzyme biosensor based on carbon nanotubes modified electrode for detection of pesticides residue

    Institute of Scientific and Technical Information of China (English)

    Shu Ping Zhang; Lian Gang Shan; Zhen Ran Tian; Yi Zheng; Li Yi Shi; Deng Song Zhang

    2008-01-01

    The paper describes a controllable layer-by-layer (LBL) self-assembly modification technique of multi-walled carbon nanotubes(MWNTs) and poly(diallyldimethylammonium chloride) (PDDA) towards glassy carbon electrode (GCE), Acetylcholinesterase(ACHE) was immobilized directly to the modified GCE by LBL self-assembly method, the activity value of AChE was detected byusing i-t technique based on the modified Ellman method. Then the composition of carbaryl were detected by the enzyme electrodewith 0.01U activity value and the detection limit of carbaryl is 10-12 g L-1 so the enzyme biosensor showed good properties forpesticides residue detection.2008 Shu Ping Zhang. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

  11. Tetranectin-binding site on plasminogen kringle 4 involves the lysine-binding pocket and at least one additional amino acid residue

    DEFF Research Database (Denmark)

    Graversen, Jonas Heilskov; Sigurskjold, B W; Thøgersen, H C

    2000-01-01

    that all amino acid residues of plasminogen kringle 4 found to be involved in t-AMCHA binding are also involved in binding tetranectin. Notably, one amino acid residue of plasminogen kringle 4, Arg 32, not involved in binding t-AMCHA, is critical for binding tetranectin. We also find that Asp 57 and Asp 55......, we analyze the interaction of wild-type and six single-residue mutants of recombinant plasminogen kringle 4 expressed in Escherichia coli with the recombinant C-type lectin domain of tetranectin and trans-aminomethyl-cyclohexanoic acid (t-AMCHA) using isothermal titration calorimetry. We find...

  12. Probing China's Lysine Market

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    @@ The lysine sector in China developed further in 2006. Both the capacity and the output hit new highs and China had a major impact on the global lysine market. The import amount of lysine satisfied only a very small portion of the domestic market's demand.

  13. Modified Shear Box Test Apparatus for Measuring Shear Strength of Unsaturated Residual Soil

    Directory of Open Access Journals (Sweden)

    Bujang B.K. Huat

    2005-01-01

    Full Text Available Residual soils occur in most countries of the world but the greater areas and depths are normally found in tropical humid areas. Most of these soils exhibit high suctions for most of the year. The shear strength parameters, c’ and Φ’, of soil can be obtained using conventional shear strength tests. However the conventional shear strength test equipments would not be able to measure Φb value (change of shear strength to change in suction without certain modification to them. This study describes the modifications that have been made to a standard shear box test apparatus to enable it to test soil samples in unsaturated conditions. The modifications include fabrication of an air pressure chamber, modifications of the shear box assembly inside the air pressure chamber, modification to the normal loading system, as well as additions of data acquisition devices to enhance the performance and simplify the usage of the modified shear box test apparatus.

  14. Use of the modified Ames test as an indicator of the carcinogenicity of residual aromatic extracts

    Energy Technology Data Exchange (ETDEWEB)

    Boogaard, P.; Hedelin, A.; Riley, A.; Rushton, E.; Vaissiere, M.; Minsavage, G.; Rohde, A.; Dalbey, W.

    2013-01-15

    Existing data demonstrate that residual aromatic extracts (RAEs) can be either carcinogenic or non-carcinogenic. CONCAWE had previously concluded that 'Although limited data available indicate that some RAEs are weakly carcinogenic, it is not possible to provide a general recommendation. Classify on a case-by-case basis' (CONCAWE 2005). Therefore CONCAWE's Health/Toxicology Subgroup (H/TSG) has developed a proposal for the use of the modified Ames test as a short-term predictive screening tool for decisions on the classification of RAEs for carcinogenicity. The relationship between RAE chemistry and carcinogenic potential is not as well understood as it is for some other categories of substances, e.g. Other Lubricant Base Oils (OLBO). However, a correlation has been found between the results of the skin carcinogenicity bioassay and the mutagenicity index (MI) obtained from the modified Ames test. Data supporting this correlation are summarised in this report. The H/TSG confirmed that the modified Ames test can be used as a predictive screening tool and that a cut-off value can be established to make a distinction between carcinogenic and non-carcinogenic products. RAEs with a MI > 0.4 demonstrated carcinogenic potential upon dermal application to mouse skin with chronic exposure. RAEs with a MI > 0.4 did not demonstrate a carcinogenic potential. To justify the use of the modified Ames test with RAEs, additional analysis of the repeatability of the test with RAEs was required. With this objective, CONCAWE sponsored a round robin study with different samples of RAEs from member companies, at three different laboratories. The repeatability demonstrated in the round robin study with RAEs support the proposed use of the modified Ames test. As part of the tools available for use by member companies, the H/TSG proposed a standard operating procedure (SOP) (included as an Appendix to this report) on the conduct of the modified Ames test with RAEs. The H

  15. An update on histone lysine methylation in plants

    Institute of Scientific and Technical Information of China (English)

    Yu Yu; Zhongyuan Bu; Wen-Hui Shen; Aiwu Dong

    2009-01-01

    Histone methylation plays crucial roles in epigenetic regulation.The SET domain proteins are now recognized as generally having methyltransferase activity targeted to specific lysine residues of histones.The enzymes and their specific histone lysine methylation have enormous impacts on the regulation of chromatin structure and function.In this review,we discuss recent advances made on histone lysine methylations and their diverse functions in plant growth and development.

  16. Slaking characteristics of unsaturated granite residual soils based on a modified slaking test method

    Science.gov (United States)

    Zhang, S.

    2012-12-01

    Slaking is one of the distinct process involved in the structural breakdown that occurs when soils are suddenly immersed in, or placed in contact with, water. The process occurs because the tensile stress of soil skeleton cannot withstand the stresses caused by rapid water uptake. Some instability problems caused by slaking process were found on subway tunnels and engineered slopes excavated in granite residual soils (GRS) in Guangzhou, south China. A serious of experimental laboratory studies were carried out in order to get better understanding about the slaking characteristics of GRS. Unsaturated GRS samples with different initial moisture content and different degree of compaction were made for test using homemade apparatus. We proposed a modified slaking test mothod to obtain slaking curves so as to reflect the actual slaking process on the basis of experimental observation and mechanism analysis as much as possible. The method considerred air escape process during water uptaking which combined the two extremes involved in water uptaking with free escape of displaced air and with no air escape. Subsequently, a modified slaking velocity index based on the the slaking curve was calculated and utilized for further data processing and analysis. We discussed the relationship between two main control factors (fillable porosity of soil and initial matric suction of soil) and slaking velocity as well. The results reveal that it has exponential function relationship for fillable porosity of soil and logarithm function relationship for initial matric suction of soil.

  17. Installation of site-specific methylation into histones using methyl lysine analogs.

    Science.gov (United States)

    Simon, Matthew D

    2010-04-01

    Chromatin structure is influenced by post-translational modifications on histones, the principal basic protein component of chromatin. In order to study one of these modifications, lysine methylation, in the context of reconstituted chromatin, this unit describes the installation of analogs of methyl lysine residues into recombinant histones. The modification site is specified by mutating the lysine of interest to cysteine. The mutant histones are expressed and purified, and the cysteine residue alkylated to produce N-methyl aminoethylcysteine, an isosteric analog of methyl lysine. Using different alkylating reagents, it is possible to install analogs of mono-, di-, or trimethyl lysine. While these analogs are not identical to methyl lysine residues, they show similar biochemical properties to their natural counterparts. The ease of synthesis of methyl lysine analog (MLA) histones, especially on a large scale, makes them particularly useful reagents for studying the effects of histone lysine methylation on chromatin structure, biophysics and biochemistry. (c) 2010 by John Wiley & Sons, Inc.

  18. MODIFIED LAYER REMOVAL METHOD FOR MEASUREMENT OF RESIDUAL STRESS DISTRIBUTION IN THICK PRE-STRETCHED ALUMINUM PLATE

    Institute of Scientific and Technical Information of China (English)

    WANGShu-hong; ZUODun-wen; WANGMin; WANGZong-rong

    2004-01-01

    The integrated structure parts are widely used in aircraft. The distortion caused by residual stresses in thick pre-stretched aluminum plates during machining integrated parts is a common and serious problem. To predict and control the machining distortion, the residual stress distribution in the thick plate must be measured firstly. The modified removal method for measuring residual stress in thick pre-stretched aluminum plates is proposed and the stress-strain relation matrix is deduced by elasticity theory. The residual stress distribution in specimen of 7050T7451 plate is measured by using the method, and measurement results are analyzed and compared with data obtained by other methods. The method is effective to measure the residual stress.

  19. SPOTing Acetyl-Lysine Dependent Interactions

    Directory of Open Access Journals (Sweden)

    Sarah Picaud

    2015-08-01

    Full Text Available Post translational modifications have been recognized as chemical signals that create docking sites for evolutionary conserved effector modules, allowing for signal integration within large networks of interactions. Lysine acetylation in particular has attracted attention as a regulatory modification, affecting chromatin structure and linking to transcriptional activation. Advances in peptide array technologies have facilitated the study of acetyl-lysine-containing linear motifs interacting with the evolutionary conserved bromodomain module, which specifically recognizes and binds to acetylated sequences in histones and other proteins. Here we summarize recent work employing SPOT peptide technology to identify acetyl-lysine dependent interactions and document the protocols adapted in our lab, as well as our efforts to characterize such bromodomain-histone interactions. Our results highlight the versatility of SPOT methods and establish an affordable tool for rapid access to potential protein/modified-peptide interactions involving lysine acetylation.

  20. Identification of critical residues of the serotype modifying O-acetyltransferase of Shigella flexneri

    Directory of Open Access Journals (Sweden)

    Thanweer Farzaana

    2012-07-01

    Full Text Available Abstract Background Thirteen serotypes of Shigella flexneri (S. flexneri have been recognised, all of which are capable of causing bacillary dysentery or shigellosis. With the emergence of the newer S. flexneri serotypes, the development of an effective vaccine has only become more challenging. One of the factors responsible for the generation of serotype diversity is an LPS O-antigen modifying, integral membrane protein known as O-acetyltransferase or Oac. Oac functions by adding an acetyl group to a specific O-antigen sugar, thus changing the antigenic signature of the parent S. flexneri strain. Oac is a membrane protein, consisting of hydrophobic and hydrophilic components. Oac bears homology to several known and predicted acetyltransferases with most homology existing in the N-terminal transmembrane (TM regions. Results In this study, the conserved motifs in the TM regions and in hydrophilic loops of S. flexneri Oac were targeted for mutagenesis with the aim of identifying the amino acid residues essential for the function of Oac. We previously identified three critical arginines–R73, R75 and R76 in the cytoplasmic loop 3 of Oac. Re-establishing that these arginines are critical, in this study we suggest a catalytic role for R73 and a structural role for R75 and R76 in O-acetylation. Serine-glycine motifs (SG 52–53, GS 138–139 and SYG 274–276, phenylalanine-proline motifs (FP 78–79 and FPV 282–84 and a tryptophan-threonine motif (WT141-142 found in TM segments and residues RK 110–111, GR 269–270 and D333 found in hydrophilic loops were also found to be critical to Oac function. Conclusions By studying the effect of the mutations on Oac’s function and assembly, an insight into the possible roles played by the chosen amino acids in Oac was gained. The transmembrane serine-glycine motifs and hydrophilic residues (RK 110–111, GR 269–270 and D333 were shown to have an affect on Oac assembly which suggests a structural role

  1. Post-voiding residual urine and capacity increase in orthotopic urinary diversion: Standard vs modified technique

    Directory of Open Access Journals (Sweden)

    Bančević Vladimir

    2010-01-01

    Full Text Available Background/Aim. Ever since the time when the first orthotopic urinary diversion (pouch was performed there has been a constant improvement and modification of surgical techniques. The aim has been to create a urinary reservoir similar to normal bladder, to decrease incidence of postoperative complications and provide an improved life quality. The aim of this study was to compare postvoiding residual urine (PVR and capacity of the pouch constructed by standard or modified technique. Methods. In this prospective and partially retrospective clinical study we included 79 patients. In the group of 41 patients (group ST pouch was constructed using 50-70 cm of the ileum (standard technique. In the group of 38 patients (group MT pouch was constructed using 25-35 cm of the ileum (modified technique. Postoperatively, PVR and pouch capacity were measured using ultrasound in a 3-, 6- and 12-month period. Results. Postoperatively, an increase in PVR and pouch capacity was noticed in both groups. Twelve months postoperatively, PVR was significantly smaller in the group MT than in the group ST [23 (0-90 mL vs 109 (0-570 mL, p < 0,001]. In the same period the pouch capacity was significantly smaller in the MT group than in the ST group [460 (290-710 mL vs 892 (480-2 050 mL, p < 0.001]. Conclusion. Postoperatively, an increase in PVR and pouch capacity was noticed during a 12-month period. A year following the operation the pouch created from a shorter ileal segment reached capacity of the 'normal' bladder with small PVR. The pouch created by standard technique developed an unnecessary large PVR and capacity.

  2. Mutational, kinetic, and NMR studies of the roles of conserved glutamate residues and of lysine-39 in the mechanism of the MutT pyrophosphohydrolase.

    Science.gov (United States)

    Harris, T K; Wu, G; Massiah, M A; Mildvan, A S

    2000-02-22

    The MutT enzyme catalyzes the hydrolysis of nucleoside triphosphates (NTP) to NMP and PP(i) by nucleophilic substitution at the rarely attacked beta-phosphorus. The solution structure of the quaternary E-M(2+)-AMPCPP-M(2+) complex indicated that conserved residues Glu-53, -56, -57, and -98 are at the active site near the bound divalent cation possibly serving as metal ligands, Lys-39 is positioned to promote departure of the NMP leaving group, and Glu-44 precedes helix I (residues 47-59) possibly stabilizing this helix which contributes four catalytic residues to the active site [Lin, J. , Abeygunawardana, C., Frick, D. N., Bessman, M. J., and Mildvan, A. S. (1997) Biochemistry 36, 1199-1211]. To test these proposed roles, the effects of mutations of each of these residues on the kinetic parameters and on the Mn(2+), Mg(2+), and substrate binding properties were examined. The largest decreases in k(cat) for the Mg(2+)-activated enzyme of 10(4.7)- and 10(2.6)-fold were observed for the E53Q and E53D mutants, respectively, while 97-, 48-, 25-, and 14-fold decreases were observed for the E44D, E56D, E56Q, and E44Q mutations, respectively. Smaller effects on k(cat) were observed for mutations of Glu-98 and Lys-39. For wild type MutT and its E53D and E44D mutants, plots of log(k(cat)) versus pH exhibited a limiting slope of 1 on the ascending limb and then a hump, i.e., a sharply defined maximum near pH 8 followed by a plateau, yielding apparent pK(a) values of 7.6 +/- 0.3 and 8.4 +/- 0.4 for an essential base and a nonessential acid catalyst, respectively, in the active quaternary MutT-Mg(2+)-dGTP-Mg(2+) complex. The pK(a) of 7.6 is assigned to Glu-53, functioning as a base catalyst in the active quaternary complex, on the basis of the disappearance of the ascending limb of the pH-rate profile of the E53Q mutant, and its restoration in the E53D mutant with a 10(1.9)-fold increase in (k(cat))(max). The pK(a) of 8.4 is assigned to Lys-39 on the basis of the disappearance

  3. System-wide Analysis of SUMOylation Dynamics in Response to Replication Stress Reveals Novel Small Ubiquitin-like Modified Target Proteins and Acceptor Lysines Relevant for Genome Stability

    DEFF Research Database (Denmark)

    Xiao, Zhenyu; Chang, Jer-Gung; Hendriks, Ivo A;

    2015-01-01

    . Following statistical analysis on five biological replicates, a total of 566 SUMO-2 targets were identified. After 2 hours of Hydroxyurea treatment, 10 proteins were up-regulated for SUMOylation and 2 proteins were down-regulated for SUMOylation, whereas after 24 hours, 35 proteins were up......-regulated for SUMOylation and 13 proteins were down-regulated for SUMOylation. A site-specific approach was used to map over 1,000 SUMO-2 acceptor lysines in target proteins. The methodology is generic and is widely applicable in the ubiquitin field. A large subset of these identified proteins function in one network...... that consists of interacting replication factors, transcriptional regulators, DNA damage response factors including MDC1, ATR-interacting protein ATRIP, the Bloom syndrome protein and the BLM-binding partner RMI1, the crossover junction endonuclease EME1, BRCA1 and CHAF1A. Furthermore, centromeric proteins...

  4. Elucidating the effects of arginine and lysine on a monoclonal antibody C-terminal lysine variation in CHO cell cultures.

    Science.gov (United States)

    Zhang, Xintao; Tang, Hongping; Sun, Ya-Ting; Liu, Xuping; Tan, Wen-Song; Fan, Li

    2015-08-01

    C-terminal lysine variants are commonly observed in monoclonal antibodies (mAbs) and found sensitive to process conditions, especially specific components in culture medium. The potential roles of media arginine (Arg) and lysine (Lys) in mAb heavy chain C-terminal lysine processing were investigated by monitoring the lysine variant levels under various Arg and Lys concentrations. Both Arg and Lys were found to significantly affect lysine variant level. Specifically, lysine variant level increased from 18.7 to 31.8 % when Arg and Lys concentrations were increased from 2 to 10 mM. Since heterogeneity of C-terminal lysine residues is due to the varying degree of proteolysis by basic carboxypeptidases (Cps), enzyme (basic Cps) level, pH conditions, and product (Arg and Lys) inhibition, which potentially affect the enzymatic reaction, were investigated under various Arg and Lys conditions. Enzyme level and pH conditions were found not to account for the different lysine variant levels, which was evident from the minimal variation in transcription level and intracellular pH. On the other hand, product inhibition effect of Arg and Lys on basic Cps was evident from the notable intracellular and extracellular Arg and Lys concentrations comparable with Ki values (inhibition constant) of basic Cps and further confirmed by cell-free assays. Additionally, a kinetic study of lysine variant level during the cell culture process enabled further characterization of the C-terminal lysine processing.

  5. 柠檬酸改性糠醛渣的制备%Preparation of Modified Furfural Residue With Citric Acid

    Institute of Scientific and Technical Information of China (English)

    王昕; 邢琦; 任广军

    2014-01-01

    The preparation process of modified furfural residue with citric acid was studied as well as the optimum conditions for modification. The furfural residue was first pretreated, then it was treated by 20%isopropanol and 20%sodium hydroxide solution, at last the furfural residue was modified by citric acid to obtain the citric acid modified furfural residue. Effects of reaction time, reaction temperature, citric acid solution concentration, ratio of solid to liquid on the modification were investigated. The results show that,the best modification conditions are as follows:reaction time 60 min, reaction temperature 80 ℃, citric acid concentration 100 g/L, ratio of solid to liquid 1︰4. The modified furfural residue has good adsorption for methylene blue solution, removal rate can reach to 98.2%.%研究了柠檬酸改性糠醛渣的制备过程和条件。首先对糠醛渣进行预处理,然后分别用20%的异丙醇和20%的氢氧化钠溶液处理,最后用柠檬酸对其进行改性,得到柠檬酸改性糠醛渣。讨论了反应时间,反应温度,柠檬酸溶液浓度,固液比等因素对改性的影响。结果表明:当反应时间60 min,反应温度80℃,柠檬酸质量浓度100 g/L,固液比1︰3时获得的改性糠醛渣吸附亚甲基蓝效果最好,去除率可达到98.2%。

  6. Effect of dietary lysine on hepatic lysine catabolism in broilers

    Science.gov (United States)

    Lysine is frequently a first- or second-limiting amino acid in poultry diets. Improving the efficiency of lysine use for protein synthesis would effectively lower the lysine requirement and decrease feed costs. Understanding how lysine is degraded and how the degradation is regulated would identif...

  7. The geranyl-modified tryptophan residue is crucial for ComXRO-E-2 pheromone biological activity.

    Science.gov (United States)

    Tsuji, Fumitada; Kobayashi, Ko; Okada, Masahiro; Yamaguchi, Hisao; Ojika, Makoto; Sakagami, Youji

    2011-07-01

    The ComX pheromone is an isoprenoidal oligopeptide containing a modified tryptophan residue, which stimulates natural genetic competence in gram-positive bacteria, Bacillus. We have reported the structure of the ComX(RO-E-2) pheromone, which is produced by the RO-E-2 strain of Bacillus subtilis. ComX(RO-E-2) analogs with substituted amino acids and isoprenoid modified tryptophan residues (e.g., prenyl, geranyl, and farnesyl), were synthesized and examined for biological activity. These results indicate that Phe-Trp(∗)(Ger)-NH(2) is the minimum pharmacophore of the ComX(RO-E-2) pheromone. Furthermore, the length of the isoprenoid moiety (i.e., modification style), and the presence of double bonds, are crucial for biological activity. The modification style of the ComX pheromone is more important than the peptide sequence with respect to biological activity.

  8. Removal of polycyclic aromatic hydrocarbons from aqueous solution by raw and modified plant residue materials as biosorbents.

    Science.gov (United States)

    Xi, Zemin; Chen, Baoliang

    2014-04-01

    Removal of polycyclic aromatic hydrocarbons (PAHs), e.g., naphthalene, acenaphthene, phenanthrene and pyrene, from aqueous solution by raw and modified plant residues was investigated to develop low cost biosorbents for organic pollutant abatement. Bamboo wood, pine wood, pine needles and pine bark were selected as plant residues, and acid hydrolysis was used as an easily modification method. The raw and modified biosorbents were characterized by elemental analysis, Fourier transform infrared spectroscopy and scanning electron microscopy. The sorption isotherms of PAHs to raw biosorbents were apparently linear, and were dominated by a partitioning process. In comparison, the isotherms of the hydrolyzed biosorbents displayed nonlinearity, which was controlled by partitioning and the specific interaction mechanism. The sorption kinetic curves of PAHs to the raw and modified plant residues fit well with the pseudo second-order kinetics model. The sorption rates were faster for the raw biosorbents than the corresponding hydrolyzed biosorbents, which was attributed to the latter having more condensed domains (i.e., exposed aromatic core). By the consumption of the amorphous cellulose component under acid hydrolysis, the sorption capability of the hydrolyzed biosorbents was notably enhanced, i.e., 6-18 fold for phenanthrene, 6-8 fold for naphthalene and pyrene and 5-8 fold for acenaphthene. The sorption coefficients (Kd) were negatively correlated with the polarity index [(O+N)/C], and positively correlated with the aromaticity of the biosorbents. For a given biosorbent, a positive linear correlation between logKoc and logKow for different PAHs was observed. Interestingly, the linear plots of logKoc-logKow were parallel for different biosorbents. These observations suggest that the raw and modified plant residues have great potential as biosorbents to remove PAHs from wastewater. Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of

  9. [The effect of the raw protein supply on the lysine requirements of young pigs of 12-40 kg. 1. Report. Feeding studies with wheat-peanut extraction residue rations].

    Science.gov (United States)

    Schüler, D; Bodenstein, K H; Hennig, A

    1976-09-01

    10 feedings trials were carried out with a total of more than 500 pigs weighing 12 to 40 kgs. To investigate the lysine needs of growing pigs, the animals were fed rations of wheat + extracted ground nut meal. Different food mixtures were tested containing 5 levels of crude protein (19%, 17%, 15%, 13% and 11% of the dry feed. Within each crude protein level 6 graded lysine supplements were added to the ration. The trial showed that the lysine requirements of the weaned pigs were in a decisive measure determined by the percentage proportion of crude protein contained in the ration. The crude protein portion may be calculated (for the liveweight range tested) by using the following regression equation: y=0.28+0.075x (y=lysine requirements expressed as % of the air-dried ration; x=percentage proportion of crude protein in the ration). Rations containing only protein sources of vegetable origin, with a minimum protein content of 15%, produced the same daily weight gains (520 g) as a standard diet, if the lysine demands were met through the supplementation of synthetic lysine.

  10. Modified Coconut Copra Residues as a Low Cost Biosorbent for Adsorption of Humic Substances from Peat Swamp Runoff

    Directory of Open Access Journals (Sweden)

    Siong Fong Sim

    2013-12-01

    Full Text Available The presence of dissolved organic matter, scientifically known as humic substances, gives an undesirable color and taste to water. In addition, they are the precursors of carcinogenic disinfection by-products upon disinfection treatment. Adsorption provides a potential means of removal of humic substances, and lignocellulosic biomass serves as a promising candidate. In this paper, we report the application of modified coconut copra residues for adsorption of humic substances from peat swamp runoff. The FTIR spectra suggest that coconut copra residues are genuinely rich with carboxyl groups with long alkyl chains; this renders the material a natural biosorbent, attaining an average of 50% removal under the conditions of testing. Upon treatment, dissolution of lignin and hemicellulose with the enhancement of effective carboxyl groups occurs, improving the adsorption efficiency to 96%; the treated water is visibly clear. The relative band abundance and band shifts further confirm the involvement of the surface functional groups in the adsorption process. The modified coconut copra residue is an attractive biosorbent option for removal of humic substances. The operating conditions are mild, involving non-toxic chemicals, and no pH adjustment is necessary to allow adsorption.

  11. ß-Lysine discrimination by lysyl-tRNA synthetase

    DEFF Research Database (Denmark)

    Gilreath, Marla S; Roy, Hervé; Bullwinkle, Tammy J

    2011-01-01

    guided by the PoxA structure. A233S LysRS behaved as wild type with a-lysine, while the G469A and A233S/G469A variants decreased stable a-lysyl-adenylate formation. A233S LysRS recognized ß-lysine better than wildtype, suggesting a role for this residue in discriminating a- and ß-amino acids. Both...

  12. Modified Mitchell osteotomy alone does not have higher rate of residual metatarsalgia than combined first and lesser metatarsal osteotomy.

    Science.gov (United States)

    Chen, Shu-Jung; Cheng, Yuh-Min; Lin, Sung-Yen; Chen, Chung-Hwan; Huang, Hsuan-Ti; Huang, Peng-Ju

    2015-04-01

    Transfer metatarsalgia (TM) is a common forefoot disorder secondary to hallux valgus (HV). Some authors suggest that a combined lesser metatarsal osteotomy while undergoing HV surgery improves metatarsalgia, whereas others concluded that isolated HV corrective osteotomy can improve symptomatic metatarsalgia. The main purpose of this retrospective study was to compare clinical outcomes in patients with and without combined lesser metatarsal osteotomy while receiving HV correction surgery. We retrospectively reviewed the patients who underwent osteotomy for HV correction between January 2000 and December 2010. All patients underwent HV correction with modified Mitchell osteotomy. Clinical evaluations including the American Orthopaedic Foot and Ankle Society score and residual metatarsalgia were assessed, and radiographic measurements were carried out. Sixty-five patients (83 feet) meeting the selection criteria were enrolled. Thirty feet receiving a combined lesser metatarsal osteotomy were classified as the combined surgery (CS) group, and the others were classified as the control (CN) group (53 feet). The overall rate of persistent symptomatic metatarsalgia was 19.28% after operative treatment. There were six feet with residual metatarsalgia in the CS group, and 10 feet in the CN group. There was no significant difference in the rate of persistent symptoms between the two groups (p = 0.9). According to this result, modified Mitchell osteotomy alone did not have a higher rate of residual metatarsalgia than CS. We also found that the average recovery rate of TM was about 80.7% and those patients whose preoperative HV angle was > 30° had the higher risk of residual metatarsalgia after surgery. Copyright © 2015. Published by Elsevier Taiwan.

  13. Modified Mitchell osteotomy alone does not have higher rate of residual metatarsalgia than combined first and lesser metatarsal osteotomy

    Directory of Open Access Journals (Sweden)

    Shu-Jung Chen

    2015-04-01

    Full Text Available Transfer metatarsalgia (TM is a common forefoot disorder secondary to hallux valgus (HV. Some authors suggest that a combined lesser metatarsal osteotomy while undergoing HV surgery improves metatarsalgia, whereas others concluded that isolated HV corrective osteotomy can improve symptomatic metatarsalgia. The main purpose of this retrospective study was to compare clinical outcomes in patients with and without combined lesser metatarsal osteotomy while receiving HV correction surgery. We retrospectively reviewed the patients who underwent osteotomy for HV correction between January 2000 and December 2010. All patients underwent HV correction with modified Mitchell osteotomy. Clinical evaluations including the American Orthopaedic Foot and Ankle Society score and residual metatarsalgia were assessed, and radiographic measurements were carried out. Sixty-five patients (83 feet meeting the selection criteria were enrolled. Thirty feet receiving a combined lesser metatarsal osteotomy were classified as the combined surgery (CS group, and the others were classified as the control (CN group (53 feet. The overall rate of persistent symptomatic metatarsalgia was 19.28% after operative treatment. There were six feet with residual metatarsalgia in the CS group, and 10 feet in the CN group. There was no significant difference in the rate of persistent symptoms between the two groups (p = 0.9. According to this result, modified Mitchell osteotomy alone did not have a higher rate of residual metatarsalgia than CS. We also found that the average recovery rate of TM was about 80.7% and those patients whose preoperative HV angle was > 30° had the higher risk of residual metatarsalgia after surgery.

  14. Systemic lupus erythematosus patients contain significantly less igm against mono-methylated lysine than healthy subjects.

    Directory of Open Access Journals (Sweden)

    Sha Guo

    Full Text Available Post-translational modifications on proteins are important in biological processes but may create neo-epitopes that induce autoimmune responses. In this study, we measured the serum IgG and IgM response to a set of non-modified or acetyl- and methyl-modified peptides corresponding to residues 1-19 of the histone 3 N-terminal tail in systemic lupus erythematosus (SLE patients and healthy subjects. Our results indicated that the SLE patients and healthy subjects produced antibodies (Abs to the peptides, but the two groups had different Ab isotype and epitope preferences. Abs to the non-modified form, H31-19, were of the IgG isotype and produced by SLE patients. They could not recognize the scrambled H31-19, which contained the same amino acid composition but a different sequence as H31-19. In comparison, healthy subjects in general did not produce IgG against H31-19. However, about 70% of the healthy subjects produced IgM Abs against mono-methylated K9 of H31-19 (H31-19K9me. Our further studies revealed that ε-amine mono-methylated lysine could completely inhibit the IgM binding to H31-19K9me, but lysine had no inhibitory effect. In addition, the IgM Abs could bind peptides containing a mono-methylated lysine residue but with totally different sequences. Thus, mono-methylated lysine was the sole epitope for the IgM. Interestingly, SLE patients had much lower levels of this type of IgM. There was no obvious correlation between the IgM levels and disease activity and the decreased IgM was unlikely caused by medical treatments.We also found that the IgM Abs were not polyreactive to dsDNA, ssDNA, lipopolysaccharide (LPS or insulin and they did not exist in umbilical cord serum, implying that they were not natural Abs. The IgM Abs against mono-methylated lysine are present in healthy subjects but are significantly lower in SLE patients, suggesting a distinct origin of production and special physiological functions.

  15. Mass spectrometric analysis of lysine ubiquitylation reveals promiscuity at site level

    DEFF Research Database (Denmark)

    Danielsen, Jannie M R; Sylvestersen, Kathrine B; Bekker-Jensen, Simon;

    2011-01-01

    The covalent attachment of ubiquitin to proteins regulates numerous processes in eukaryotic cells. Here we report the identification of 753 unique lysine ubiquitylation sites on 471 proteins using higher-energy collisional dissociation on the LTQ Orbitrap Velos. In total 5756 putative ubiquitin...... substrates were identified. Lysine residues targeted by the ubiquitin-ligase system show no unique sequence feature. Surface accessible lysine residues located in ordered secondary regions, surrounded by smaller and positively charged amino acids are preferred sites of ubiquitylation. Lysine ubiquitylation...

  16. Structural basis for the site-specific incorporation of lysine derivatives into proteins.

    Directory of Open Access Journals (Sweden)

    Veronika Flügel

    Full Text Available Posttranslational modifications (PTMs of proteins determine their structure-function relationships, interaction partners, as well as their fate in the cell and are crucial for many cellular key processes. For instance chromatin structure and hence gene expression is epigenetically regulated by acetylation or methylation of lysine residues in histones, a phenomenon known as the 'histone code'. Recently it was shown that these lysine residues can furthermore be malonylated, succinylated, butyrylated, propionylated and crotonylated, resulting in significant alteration of gene expression patterns. However the functional implications of these PTMs, which only differ marginally in their chemical structure, is not yet understood. Therefore generation of proteins containing these modified amino acids site specifically is an important tool. In the last decade methods for the translational incorporation of non-natural amino acids using orthogonal aminoacyl-tRNA synthetase (aaRS:tRNAaaCUA pairs were developed. A number of studies show that aaRS can be evolved to use non-natural amino acids and expand the genetic code. Nevertheless the wild type pyrrolysyl-tRNA synthetase (PylRS from Methanosarcina mazei readily accepts a number of lysine derivatives as substrates. This enzyme can further be engineered by mutagenesis to utilize a range of non-natural amino acids. Here we present structural data on the wild type enzyme in complex with adenylated ε-N-alkynyl-, ε-N-butyryl-, ε-N-crotonyl- and ε-N-propionyl-lysine providing insights into the plasticity of the PylRS active site. This shows that given certain key features in the non-natural amino acid to be incorporated, directed evolution of this enzyme is not necessary for substrate tolerance.

  17. Simultaneous determination of trace Cd(II), Pb(II) and Cu(II) by differential pulse anodic stripping voltammetry using a reduced graphene oxide-chitosan/poly-l-lysine nanocomposite modified glassy carbon electrode.

    Science.gov (United States)

    Guo, Zhuo; Li, Dong-di; Luo, Xian-Ke; Li, Ya-Hui; Zhao, Qi-Nai; Li, Meng-Meng; Zhao, Yang-Ting; Sun, Tian-Shuai; Ma, Chi

    2017-03-15

    The reduced graphene oxide (RGO) and Chitosan (CS) hybrid matrix RGO-CS were coated onto the glassy carbon electrode (GCE) surface, then, poly-l-lysine films (PLL) were prepared by electropolymerization with cyclic voltammetry (CV) method to prepare RGO-CS/PLL modified glassy carbon electrode (RGO-CS/PLL/GCE) for the simultaneous electrochemical determination of heavy metal ions Cd(II), Pb(II) and Cu(II). Combining the advantageous features of RGO and CS, RGO and CS are used together because the positively charged CS can interact with the negatively changed RGO to prevent their aggregation. Furthermore, CS has many amino groups along its macromolecular chains and possessed strongly reactive with metal ions. Moreover, PLL modified electrodes have good stability, excellent permselectivity, more active sites and strong adherence to electrode surface, which enhanced electrocatalytic activity. The RGO-CS/PLL/GCE was characterized voltammetrically using redox couples (Fe(CN)6(3-/4-)), complemented with electrochemical impedance spectroscopy (EIS). Differential pulse anodic stripping voltammetry (DPASV) has been used for the detection of Cd(II), Pb(II) and Cu(II). The detection limit of RGO-CS/PLL/GCE toward Cd(II), Pb(II) and Cu(II) is 0.01μgL(-1), 0.02μgL(-1) and 0.02μgL(-1), respectively. The electrochemical parameters that exert influence on deposition and stripping of metal ions, such as supporting electrolytes, pH value, deposition potential, and deposition time, were carefully studied.

  18. Sex Modifies Genetic Effects on Residual Variance in Urinary Calcium Excretion in Rat (Rattus norvegicus)

    OpenAIRE

    Perry, Guy M. L.; Nehrke, Keith W.; Bushinsky, David A; Reid, Robert; Lewandowski, Krista L.; Hueber, Paul; Scheinman, Steven J.

    2012-01-01

    Conventional genetics assumes common variance among alleles or genetic groups. However, evidence from vertebrate and invertebrate models suggests that residual genotypic variance may itself be under partial genetic control. Such a phenomenon would have great significance: high-variability alleles might confound the detection of “classically” acting genes or scatter predicted evolutionary outcomes among unpredicted trajectories. Of the few works on this phenomenon, many implicate sex in some a...

  19. Earthworms and Plant Residues Modify Nematodes in Tropical Cropping Soils (Madagascar: A Mesocosm Experiment

    Directory of Open Access Journals (Sweden)

    Cécile Villenave

    2010-01-01

    Full Text Available Free-living nematodes present several characteristics that have led to their use as bioindicators of soil quality. Analyzing the structure of nematofauna is a pertinent way to understand soil biological processes. Earthworms play an important role in soil biological functioning and organic matter dynamics. Their effects on soil nematofauna have seldom been studied. We studied the effect of the tropical endogeic earthworm, Pontoscolex corethrurus, on nematode community structure in a 5-month field mesocosm experiment conducted in Madagascar. Ten different treatments with or without earthworms and with or without organic residues (rice, soybean were compared. Organic residues were applied on the soil surface or mixed with the soil. The abundance of nematodes (bacterial and fungal feeders was higher in presence of P. corethrurus than in their absence. The type of plant residues as well as their localisation had significant effects on the abundance and composition of soil nematodes. The analysis of nematode community structure showed that earthworm activity led to an overall activation of the microbial compartment without specific stimulation of the bacterial or fungal compartment.

  20. Impact of dry heating on physicochemical properties of corn starch and lysine mixture.

    Science.gov (United States)

    Ji, Ying; Yu, Jicheng; Xu, Yongbin; Zhang, Yinghui

    2016-10-01

    Corn starch was modified with lysine by dry heat treatment and to investigate how they can affect the pasting and structural properties of the treated starches. Dry heating with lysine reduced the pasting temperature and resulting in viscosity increase. The particle size of heated starch-lysine mixture increased, suggesting that starch granules were cross-linked to lysine. After dry heating, the onset temperature, peak temperature and conclusion temperature of corn starch-lysine mixture were lower than those of other starches. The degree of crystallinity decreased for the starch after dry heat treatment while these heated starch samples still have the same X-ray diffraction types as the original starch.

  1. Cryptic residual GALT activity is a potential modifier of scholastic outcome in school age children with classic galactosemia.

    Science.gov (United States)

    Ryan, Emily L; Lynch, Mary Ellen; Taddeo, Elles; Gleason, Tyler J; Epstein, Michael P; Fridovich-Keil, Judith L

    2013-11-01

    Classic galactosemia is a potentially lethal disorder that results from profound deficiency of galactose-1-phosphate uridylyltransferase (GALT), the second enzyme in the Leloir pathway of galactose metabolism. Although early diagnosis and rigorous dietary restriction of galactose prevent or resolve the potentially lethal acute symptoms, patients are at markedly increased risk of long-term complications including significant cognitive, speech, and behavioral difficulties, among other problems. The mechanisms that underlie these long-term complications remain unclear, as do the factors that modify their severity. Here we explored the scholastic and behavioral outcomes experienced by a cohort of 54 school age children with classic galactosemia. Data collected included survey responses from parents and teachers, school records including standardized test scores, and GALT genotype data used to estimate predicted residual GALT activity based on a yeast expression system. As expected, many but not all of the children in our study demonstrated speech, scholastic, and behavioral difficulties. Perhaps most striking, we found that predicted cryptic residual GALT activity, often below the threshold of detection of clinical assays, appeared to modify scholastic outcome. These data raise the intriguing possibility that cryptic GALT activity might also influence the severity of other long-term complications in classic galactosemia.

  2. Bromopyruvate, an active site-directed inactivator of E. coli 2-keto-4-hydroxyglutarate(KHG) aldolase, modifies glutamic acid residue-45

    Energy Technology Data Exchange (ETDEWEB)

    Vlahos, C.J.; Dekker, E.E.

    1987-05-01

    E. coli KHG-aldolase (2-keto-4-hydroxyglutarate in equilibrium pyruvate + glyoxylate), a novel trimeric Class I aldolase, requires one active-site lysine residue (Lys 133)/subunit for Schiff-base formation as well as one arginine residue (Arg 49)/subunit for catalytic activity. The substrate analog, 3-bromopyruvate (BRPY), causes a time- and concentration-dependent loss of KHG-aldolase activity. This inactivation is regarded as active site-directed since: (a) BRPY modification results in complete loss of enzymatic activity; (b) saturation kinetics are exhibited, suggesting that a reversible complex is formed between the aldolase and BRPY prior to the rate-limiting inactivation step; (c) over 90% of the initial aldolase activity is protected by either substrate, pyruvate or KHG; (d) 1.1 mol of /sup 14/C-BRPY is bound/enzyme subunit. Peptide isolation and sequencing show that the incorporated radioactivity is associated with residue Glu-45. Denaturation of the enzyme with guanidine x HCl following treatment with excess /sup 14/C-BRPY allows for the incorporation of carbon-14 at Cys-159 and Cys-180 as well. The presence of pyruvate protects Glu-45 from being esterified but does not prevent the alkylation of the two cysteine residues. These results suggest that Glu-45 is essential for the catalytic activity of E. coli KHG-aldolase, most likely functioning as the active-site amphoteric proton donor/acceptor moiety that is involved in the overall mechanism of the reaction catalyzed by this enzyme.

  3. A lysine-to-arginine mutation on NEDD8 markedly reduces the activity of cullin RING E3 ligase through the impairment of neddylation cascades

    Energy Technology Data Exchange (ETDEWEB)

    Sui, Yiyan; Liu, Yaobin; Xu, Guoqiang, E-mail: gux2002@suda.edu.cn

    2015-06-12

    Neural-precursor-cell-expressed developmentally down-regulated 8 (NEDD8) is a ubiquitin-like modifier, which forms covalent conjugates on lysines of its substrates. This post-translational modification, neddylation, plays important roles in tumor cell proliferation and viability. Ubiquitin can form diverse polyubiquitin chains, on its seven lysines, which play important functions in various biological processes. However, the roles of lysines in NEDD8 have not been explored. Here, we generated nine NEDD8 point mutants, each with one lysine replaced by an arginine, to study the putative function of lysines in NEDD8. Our experiments discover that Lys27 in NEDD8 is a critical residue for protein neddylation. Replacement of this residue with arginine almost completely eliminates the conjugation of NEDD8 to its substrates. Furthermore, we find that the K27R mutant impairs NEDD8 conjugation to the E2 enzyme, which normally forms thioester bonds for further transferring NEDD8 to its ligases and substrates. Therefore, this mutation completely inhibits global protein neddylation, including neddylation of cullin family proteins, resulting in decreased activity of cullin-RING E3 ligases. This work sheds new light on the roles of NEDD8 lysines on neddylation cascades and provides a dominant negative mutant for the study of neddylation and its biological functions. - Highlights: • Lys27 in NEDD8 is critical for protein neddylation. • NEDD8 K27R mutant impairs the NEDD8 conjugation. • NEDD8 K27R mutant significantly reduces the activity of cullin-RING E3 ligases.

  4. Lysine acetylation is a common post-translational modification of key metabolic pathway enzymes of the anaerobe Porphyromonas gingivalis.

    Science.gov (United States)

    Butler, Catherine A; Veith, Paul D; Nieto, Matthew F; Dashper, Stuart G; Reynolds, Eric C

    2015-10-14

    Porphyromonas gingivalis is a Gram-negative anaerobe considered to be a keystone pathogen in the development of the bacterial-associated inflammatory oral disease chronic periodontitis. Although post-translational modifications (PTMs) of proteins are commonly found to modify protein function in eukaryotes and prokaryotes, PTMs such as lysine acetylation have not been examined in P. gingivalis. Lysine acetylation is the addition of an acetyl group to a lysine which removes this amino acid's positive charge and can induce changes in a protein's secondary structure and reactivity. A proteomics based approach combining immune-affinity enrichment with high sensitivity Orbitrap mass spectrometry identified 130 lysine acetylated peptides from 92 P. gingivalis proteins. The majority of these peptides (71) were attributed to 45 proteins with predicted metabolic activity; these proteins could be mapped to several P. gingivalis metabolic pathways where enzymes catalysing sequential reactions within the same pathway were often found acetylated. In particular, the catabolic pathways of complex anaerobic fermentation of amino acids to produce energy had 12 enzymes lysine acetylated. The results suggest that lysine acetylation may be an important mechanism in metabolic regulation in P. gingivalis, which is vital for P. gingivalis survival and adaptation of its metabolism throughout infection. Statement of significance. Porphyromonas gingivalis is a keystone pathogen in the development of chronic periodontitis, an inflammatory disease of the supporting tissues of the teeth. The ability of the pathogen to induce dysbiosis and disease is related to an array of specific virulence factors and metabolic regulation that enables the bacterium to proliferate in an inflamed periodontal pocket. The mechanisms P. gingivalis uses to adapt to a changing and hostile environment are poorly understood and here we show, for the first time, that enzymes of critical metabolic pathways for energy

  5. Nanocomposites of polyamide 6/residual monomer with organic-modified montmorillonite and their nanofibers produced by electrospinning

    Directory of Open Access Journals (Sweden)

    Cesar Augusto Gonçalves Beatrice

    2012-08-01

    Full Text Available Nanocomposites of an organic-modified montmorillonite (MMT and polyamide 6 (PA6 with a residual monomer were produced by melt mixing in a torque rheometer. By wide angle X-rays diffraction (WAXD, intercalated/exfoliated structures were observed in the PA6/MMT nanocomposites with 3 and 5 wt. (% of MMT; on the other hand, when 7 wt. (% of MMT was added, a nanocomposite with exfoliated structures was obtained due to the predominant linking reactions between the residual monomer and the "nanoclays" organic surfactant. Solutions of these PA6/MMT nanocomposites at 15, 17 and 20 wt. (% in formic acid were prepared. The 3 and 5 wt. (% nanocomposites were successfully electrospun; however, electrospinning of the 7 wt. (% nanocomposite was not possible. WAXD, scanning and transmission electron microscopy results showed that the 3 and 5 wt. (% nanofibers with average diameter between 80-250 nm had exfoliated structures. These results indicate that the high elongational forces developed during the electrospinning process changed the initial intercalated/exfoliated structure of the nanocomposites to an exfoliated one.

  6. Multi-pesticides residue analysis of grains using modified magnetic nanoparticle adsorbent for facile and efficient cleanup.

    Science.gov (United States)

    Liu, Zhenzhen; Qi, Peipei; Wang, Xiangyun; Wang, Zhiwei; Xu, Xiahong; Chen, Wenxue; Wu, Liyu; Zhang, Hu; Wang, Qiang; Wang, Xinquan

    2017-09-01

    A facile, rapid sample pretreatment method was developed based on magnetic nanoparticles for multi-pesticides residue analysis of grains. Magnetite (Fe3O4) nanoparticles modified with 3-(N,N-diethylamino)propyltrimethoxysilane (Fe3O4-PSA) and commercial C18 were selected as the cleanup adsorbents to remove the target interferences of the matrix, such as fatty acids and non-polar compounds. Rice was used as the representative grain sample for method optimization. The amount of Fe3O4-PSA and C18 were systematically investigated for selecting the suitable purification conditions, and the simultaneous determination of 50 pesticides and 8 related metabolites in rice was established by liquid chromatography-tandem mass spectrometry. Under the optimal conditions, the method validation was performed including linearity, sensitivity, matrix effect, recovery and precision, which all satisfy the requirement for pesticides residue analysis. Compared to the conventional QuEChERS method with non-magnetic material as cleanup adsorbent, the present method can save 30% of the pretreatment time, giving the high throughput analysis possible. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Mutation in aspartic acid residues modifies catalytic and haemolytic activities of Bacillus cereus sphingomyelinase.

    Science.gov (United States)

    Tamura, H; Tameishi, K; Yamada, A; Tomita, M; Matsuo, Y; Nishikawa, K; Ikezawa, H

    1995-01-01

    Four aspartic acid residues (Asp126, Asp156, Asp233 and Asp295) of Bacillus cereus sphingomyelinase (SMase) in the conservative regions were changed to glycine by in vitro mutagenesis, and the mutant SMases [D126G (Asp126-->Gly etc.), D156G, D233G and D295G] were produced in Bacillus brevis 47, a protein-producing strain. The sphingomyelin (SM)-hydrolysing activity of D295G was completely abolished and those of D126G and D156G were reduced by more than 80%, whereas that of D233G was not so profoundly affected. Two mutant enzymes (D126G and D156G) were purified and characterized further. The hydrolytic activities of D126G and D156G toward four phosphocholine-containing substrates with different hydrophobicities, SM, 2-hexadecanoylamino-4-nitrophenylphosphocholine(HNP), lysophosphatidylcholine (lysoPC) and p-nitro-phenylphosphocholine (p-NPPC), were compared with those of the wild-type. The activity of D126G toward water-soluble p-NPPC was comparable with that of the wild-type. On the other hand, D156G catalysed the hydrolysis of hydrophilic substrates such as HNP and p-NPPC more efficiently (> 4-fold) than the wild-type. These results suggested that Asp126 and Asp156, located in the highly conserved region, may well be involved in a substrate recognition process rather than catalytic action. Haemolytic activities of the mutant enzymes were found to be parallel with their SM-hydrolysing activities. Two regions, including the C-terminal region containing Asp295, were found to show considerable sequence identity with the corresponding regions of bovine pancreatic DNase I. Structural predictions indicated structural similarity between SMase and DNase I. An evolutionary relationship based on the catalytic function was suggested between the structures of these two phosphodiesterases. Images Figure 2 Figure 3 Figure 4 Figure 6 PMID:7639690

  8. Chemical tools for unraveling the substrate specificity of the lysine deacylase enzymes

    DEFF Research Database (Denmark)

    Madsen, Andreas Stahl; Olsen, Christian Adam

    The lysine deacylase (KDAC) enzymes catalyze hydrolytic removal of acyl functionalities from theε-amino group of lysine residues ina variety of proteins including histones, and KDAC-mediated deacetylation of proteins has been established as a key epigeneticandmetabolic regulator. Recent studies h......-dependent HDACs 1–11 as well as NAD + -dependent sirtuins (SIRT1–7) will be discussed....

  9. New modified multi-level residue harmonic balance method for solving nonlinearly vibrating double-beam problem

    Science.gov (United States)

    Rahman, Md. Saifur; Lee, Yiu-Yin

    2017-10-01

    In this study, a new modified multi-level residue harmonic balance method is presented and adopted to investigate the forced nonlinear vibrations of axially loaded double beams. Although numerous nonlinear beam or linear double-beam problems have been tackled and solved, there have been few studies of this nonlinear double-beam problem. The geometric nonlinear formulations for a double-beam model are developed. The main advantage of the proposed method is that a set of decoupled nonlinear algebraic equations is generated at each solution level. This heavily reduces the computational effort compared with solving the coupled nonlinear algebraic equations generated in the classical harmonic balance method. The proposed method can generate the higher-level nonlinear solutions that are neglected by the previous modified harmonic balance method. The results from the proposed method agree reasonably well with those from the classical harmonic balance method. The effects of damping, axial force, and excitation magnitude on the nonlinear vibrational behaviour are examined.

  10. Extensive lysine methylation in hyperthermophilic crenarchaea : potential implications for protein stability and recombinant enzymes

    OpenAIRE

    Botting, Catherine H.; Paul Talbot; Sonia Paytubi; White, Malcolm F

    2010-01-01

    In eukarya and bacteria, lysine methylation is relatively rare and is catalysed by sequence-specific lysine methyltransferases that typically have only a single-protein target. Using RNA polymerase purified from the thermophilic crenarchaeum Sulfolobus solfataricus, we identified 21 methyllysines distributed across 9 subunits of the enzyme. The modified lysines were predominantly in alpha-helices and showed no conserved sequence context. A limited survey of the Thermoproteus tenax proteome re...

  11. Characterization and crystal structure of lysine insensitive Corynebacterium glutamicum dihydrodipicolinate synthase (cDHDPS) protein.

    Science.gov (United States)

    Rice, Elena A; Bannon, Gary A; Glenn, Kevin C; Jeong, Soon Seog; Sturman, Eric J; Rydel, Timothy J

    2008-12-15

    The lysine insensitive Corynebacterium glutamicum dihydrodipicolinate synthase enzyme (cDHDPS) was recently successfully introduced into maize plants to enhance the level of lysine in the grain. To better understand lysine insensitivity of the cDHDPS, we expressed, purified, kinetically characterized the protein, and solved its X-ray crystal structure. The cDHDPS enzyme has a fold and overall structure that is highly similar to other DHDPS proteins. A noteworthy feature of the active site is the evidence that the catalytic lysine residue forms a Schiff base adduct with pyruvate. Analyses of the cDHDPS structure in the vicinity of the putative binding site for S-lysine revealed that the allosteric binding site in the Escherichia coli DHDPS protein does not exist in cDHDPS due to three non-conservative amino acids substitutions, and this is likely why cDHDPS is not feedback inhibited by lysine.

  12. Separating nano graphene oxide from the residual strong-acid filtrate of the modified Hummers method with alkaline solution

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Xuebing, E-mail: xuebinghu2010@gmail.com [Key Laboratory of Inorganic Membrane, Jingdezhen Ceramic Institute, Jingdezhen 333001 (China); Key Laboratory of Inorganic Coating Materials, Shanghai Institute of Ceramics, Chinese Academy of Science, Shanghai 201800 (China); Yu, Yun, E-mail: yunyush@mail.sic.ac.cn [Key Laboratory of Inorganic Coating Materials, Shanghai Institute of Ceramics, Chinese Academy of Science, Shanghai 201800 (China); Wang, Yongqing; Zhou, Jianer [Key Laboratory of Inorganic Membrane, Jingdezhen Ceramic Institute, Jingdezhen 333001 (China); Song, Lixin [Key Laboratory of Inorganic Coating Materials, Shanghai Institute of Ceramics, Chinese Academy of Science, Shanghai 201800 (China)

    2015-02-28

    Graphical abstract: By adding an alkaline (NaOH or KOH) solution, the unprecipitated nano graphene oxide undergoes fast aggregation from the residual strong-acid filtrate of the modified Hummers method and forms the stable floccules when the pH value of the filtrate is about 1.7. The acid–base interaction with the surface functional groups of the carbon layers plays a role in the aggregation of the unprecipitated nano graphene oxide. - Highlights: • The novel and high-efficient method for separating graphene oxide was showed. • Graphene oxide undergoes aggregation and forms the floccules when pH value is ∼1.7. • The acid–base interaction plays a role in the aggregation of graphene oxide. - Abstract: In the modified Hummers method for preparing graphene oxide, the yellow slurry can be obtained. After filtering through a quantitative filter paper, the strong-acid filtrate containing the unprecipitated nano graphene oxide was gained. The corresponding filtrate was added gradually with an alkaline (NaOH or KOH) solution at room temperature. The unprecipitated nano graphene oxide could undergo fast aggregation when the pH value of the filtrate was about 1.7 and formed the stable floccules. X-ray diffraction analysis shows the dominant peak of the floccules is about 11°, which accords to the peak of graphene oxide. Spectra of X-ray photoelectron spectroscopy confirm the presence in the floccules of an abundance of oxygen functional groups and the purified graphene oxide floccules can be obtained. Atomic force microscopy measurement shows the graphene oxide floccules consists of sheet-like objects, mostly containing only a few layers (about 5 layers). Zeta potential analysis demonstrates the surface charge of the graphene oxide is pH-sensitive and its isoelectric point is ∼1.7. The flocculation mechanism of graphene oxide ascribes to the acid–base interaction with the surface functional groups of the carbon layers.

  13. Induction by fructose force-feeding of histone H3 and H4 acetylation at their lysine residues around the Slc2a5 gene and its expression in mice.

    Science.gov (United States)

    Honma, Kazue; Mochizuki, Kazuki; Goda, Toshinao

    2013-01-01

    It has been reported that fructose force-feeding rapidly induced jejunal Slc2a5 gene expression in rodents. We demonstrate in this study that acetylation at lysine (K) 9 of histone H3 and acetylation at K5 and K16 of histone H4 were more enhanced in the promoter/enhancer to transcribed regions of the Slc2a5 gene in fructose force-fed mice than in glucose force-fed mice. However, fructose force-feeding did not induce acetylation at K14 of histone H3, or at K8 and K12 of histone H4 around the Slc2a5 gene. These results suggest that fructose force-feeding induced selective histone acetylation, particularly of H3 and H4, around the jejunal Slc2a5 gene in mice.

  14. Lysine Ubiquitination and Acetylation of Human Cardiac 20S Proteasomes

    Science.gov (United States)

    Lau, Edward; Choi, Howard JH; Ng, Dominic CM; Meyer, David; Fang, Caiyun; Li, Haomin; Wang, Ding; Zelaya, Ivette M; Yates, John R; Lam, Maggie PY

    2016-01-01

    Purpose Altered proteasome functions are associated with multiple cardiomyopathies. While the proteasome targets poly-ubiquitinated proteins for destruction, it itself is modifiable by ubiquitination. We aim to identify the exact ubiquitination sites on cardiac proteasomes and examine whether they are also subject to acetylations. Experimental design Assembled cardiac 20S proteasome complexes were purified from five human hearts with ischemic cardiomyopathy, then analyzed by high-resolution MS to identify ubiquitination and acetylation sites. We developed a library search strategy that may be used to complement database search in identifying PTM in different samples. Results We identified 63 ubiquitinated lysines from intact human cardiac 20S proteasomes. In parallel, 65 acetylated residues were also discovered, 39 of which shared with ubiquitination sites. Conclusion and clinical relevance This is the most comprehensive characterization of cardiac proteasome ubiquitination to-date. There are significant overlaps between the discovered ubiquitination and acetylation sites, permitting potential crosstalk in regulating proteasome functions. The information presented here will aid future therapeutic strategies aimed at regulating the functions of cardiac proteasomes. PMID:24957502

  15. New lysine-acetylated proteins screened by immunoaffinity and liquid chromatography-mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The lack of selective extraction specific for lysine-acetylated proteins has been a major problem in the field of acetylation biology,though acetylation plays a key role in many biological processes.In this paper,we report for the first time the proteomic screening of lysine-acetylated proteins from a mouse liver tissue,by a new approach of immunoaffinity purification of lysine-acetylated peptides combined with nano-HPLC/MS/MS analysis.We have found 20 lysine-acetylated proteins with 21 lysine-acetylated sites,among which 12 lysine-acetylated proteins and 16 lysine-acetylated sites have never been reported before.Notably,three acetyltransferases harboring in mitochondrion are newly discovered acetyltransferases responsible for the acetylation of nonhistone proteins.We have explored the significant patterns of residue preference by the hierarchical clustering analysis of amino acid residues surrounding acetylation sites,which could be helpful to the prediction of new sites of lysine acetylation.Our findings provide more candidates for studying the important roles played by acetylation in diverse cellular pathways and related human diseases.

  16. Effects of fortified lysine on the amino acid profile and sensory qualities of deep-fried and dried noodles.

    Science.gov (United States)

    Polpuech, C; Chavasit, V; Srichakwal, P; Paniangvait, P

    2011-08-01

    Lysine fortification of wheat flour has been used toward reducing protein energy malnutrition in developing countries. The feasibility of fortifying instant noodles with lysine was evaluated based on sensory qualities and the residual lysine content. Fifty grams of deep-fried and dried instant noodles were fortified with 0.23 and 0.21 g lysine, respectively. The production temperatures used for deep-frying were 165-175 degrees C and for drying, 80-105 degrees C; these are the temperatures used in the industrial production of both kinds of noodles. Lysine fortification was then performed at the local factories using the commercial production lines and packaging for both types of instant noodles. Both fortified and unfortified deep-fried and dried instant noodles were stored at 50 degrees C under fluorescent light for 2 and 4 months, respectively. The fortified products were tested for residual lysine content and sensory qualities as compared with unfortified noodles. The results show fortified products from the tested processing temperatures were all accepted. After storage, significant losses of lysine were not found in both types of noodles analysed. The lysine-fortified noodles had amino acid scores of 102% and 122%, respectively. After 2 months, the sensory quality of fortified deep-fried noodles was still acceptable; however, the dried noodles turned to an unacceptable dark colour. This study shows that it is feasible to fortify deep-fried instant noodles with lysine, though lysine fortification exhibited an undesirable colour in the dried instant noodles after storage.

  17. Effect of heat damage in an autoclave on the reactive lysine contents of soy products and corn distillers dried grains with solubles. Use of the results to check on lysine damage in common qualities of these ingredients.

    Science.gov (United States)

    Fontaine, Johannes; Zimmer, Ulrike; Moughan, Paul J; Rutherfurd, Shane M

    2007-12-26

    The suitability of the homoarginine reaction for determining the reactive lysine in soy products and corn distillers dried grain with solubles (DDGS) was tested. For this purpose, some batches were subjected to deliberate heat damage for up to 30 min in an autoclave with 135 degrees C hot steam, and the samples were analyzed for total lysine and reactive lysine. In addition, 84 samples of common soy and 80 samples of corn DDGS were tested for their content of total and reactive lysine, and the contents were compared with those of the autoclave tests. For soy products conclusive results were obtained. In the case of heat treatment, both total lysine and reactive lysine decrease, but the latter is clearly a more sensitive indicator of lysine damage. Most normal products are quite similar, with toasting-induced damage to reactive lysine of ca. 15% compared to untoasted beans. The cause of the constantly occurring residual lysine after guanidination and the poorer reaction balance in the case of damage were explained. For common DDGS samples, however, less favorable results were obtained. Reactive and total lysine decreased almost in parallel due to heat damage, showing a great gap between them. Results showed indeed that variation of total and reactive lysine in DDGS is high, proving that its production conditions are not yet optimal for a feed ingredient.

  18. Post-translational serine/threonine phosphorylation and lysine acetylation: a novel regulatory aspect of the global nitrogen response regulator GlnR in S. coelicolor M145.

    Directory of Open Access Journals (Sweden)

    Rafat Amin

    2016-08-01

    Full Text Available Soil-dwelling Streptomyces bacteria such as S. coelicolor have to constantly adapt to the nitrogen (N availability in their habitat. Thus, strict transcriptional and post-translational control of the N-assimilation is fundamental for survival of this species. GlnR is a global response regulator that controls transcription of the genes related to the N-assimilation in S. coelicolor and other members of the Actinomycetales. GlnR represents an atypical orphan response regulator that is not activated by the phosphorylation of the conserved aspartate residue (Asp 50. We have applied transcriptional analysis, LC-MS/MS analysis and electrophoretic mobility shift assays (EMSAs to understand the regulation of GlnR in S. coelicolor M145. The expression of glnR and GlnR-target genes was revisited under four different N-defined conditions and a complex N-rich condition. Although, the expression of selected GlnR-target genes was strongly responsive to changing N-concentrations, the glnR expression itself was independent of the N-availability. Using LC-MS/MSanalysis we demonstrated that GlnR was post-translationally modified. The post-translational modifications of GlnR comprise phosphorylation of the serine/threonine residues and acetylation of lysine residues. In the complex N-rich medium GlnR was phosphorylated on six serine/ threonine residues and acetylated on one lysine residue. Under defined N-excess conditions only two phosphorylated residues were detected whereas under defined N-limiting conditions no phosphorylation was observed. GlnR phosphorylation is thus clearly correlated with N-rich conditions. Furthermore, GlnR was acetylated on four lysine residues independently of the N-concentration in the defined media and on only one lysine residue in the complex N-rich medium. Using EMSAs we demonstrated that phosphorylation inhibited the binding of GlnR to its targets genes, whereas acetylation had little influence on the formation of GlnR-DNA complex

  19. Preparation of Modified Furfural Residue With Epoxy Chloropropane%环氧氯丙烷改性糠醛渣的制备

    Institute of Scientific and Technical Information of China (English)

    张春丽; 邢琦; 任广军

    2013-01-01

    The optimum preparation process and conditions of modified furfural residue were studied. The furfural slag was firstly pretreated, and then it was stirred in 20%sodium hydroxide solution for 30 min and laid for 18 h, at last epoxy chloropropane was added to prepare the modified furfural residue. Effects of reaction time, reaction temperature, solution concentration, ratio of solid to liquid on properties of modified furfural residue were investigated. The results show that, the best modification conditions are as follows: reaction time 30min, reaction temperature 60 ℃, epichlorohydrin concentration 70%, ratio of solid to liquid 1∶4;When the modified furfural residue is used to adsorb methylene blue solution, the removal rate can reach to 98%.%研究了改性糠醛渣的制备过程和最佳改性条件。首先将糠醛渣进行预处理,然后在浓度为20%的氢氧化钠溶液中搅拌30 min,并静置18 h,再加入环氧氯丙烷,在一定条件下进行改性得到环氧氯丙烷改性糠醛渣。再根据反应时间,反应温度,溶液浓度,固液比确定最佳改性条件,结果表明:最佳改性条件为反应时间30 min,反应温度60℃,环氧氯丙烷浓度70%,固液比1︰4。改性后的糠醛渣吸附亚甲基蓝溶液时,去除率可达到98%。

  20. The effect of modifying rooting depths and nitrification inhibitors on nutrient uptake from organic biogas residues in maize

    Science.gov (United States)

    Dietrich, Charlotte C.; Koller, Robert; Nagel, Kerstin A.; Schickling, Anke; Schrey, Silvia D.; Jablonowski, Nicolai D.

    2017-04-01

    shallower layers, where their effect on plant growth was temporarily most pronounced. At final harvest (21 DAS) however, effects of nitrification inhibitors on plant height were visible only in deeper layers. Furthermore, the statistically significant interaction between the factors time x layer depths x nitrification inhibitors underlined the dynamic influence of nitrification inhibitors on plant growth over time and across rooting depths. This study offers insights into optimizing nutrient uptake and plant productivity by (re-) using residues from the biogas industry. It is among the first to monitor and try to explain the dynamics of nitrification inhibitors on root system architecture over time. A modified N-fertilization application scheme might also serve as a promising tool in optimizing phytoremediation and phytomining techniques through predictably altering root structure in fertilized layers. References: Nagel, K. A. ; Putz, A. ; Gilmer, F. ; Heinz, K. ; Fischbach, A. ; Pfeifer, J. ; Faget, M. ; Blossfeld, S. ; Ernst, M. ; Dimaki, C. ; Kastenholz, B. ; Kleinert, A.-K. ; Galinski, A. ; Scharr, H. ; Fiorani, F. ; Schurr, U. (2012): GROWSCREEN-Rhizo is a novel phenotyping robot enabling simultaneous measurements of root and shoot growth for plants grown in soil-filled rhizotrons.
Functional plant biology 39(11), 891-904.

  1. The presence of modifiable residues in the core peptide part of precursor nisin is not crucial for precursor nisin interactions with NisB- and NisC.

    Directory of Open Access Journals (Sweden)

    Rustem Khusainov

    Full Text Available Precursor nisin is a model posttranslationally modified precursor lantibiotic that can be structurally divided into a leader peptide sequence and a modifiable core peptide part. The nisin core peptide clearly plays an important role in the precursor nisin-nisin modification enzymes interactions, since it has previously been shown that the construct containing only the nisin leader sequence is not sufficient to pull-down the nisin modification enzymes NisB and NisC. Serines and threonines in the core peptide part are the residues that NisB specifically dehydrates, and cysteines are the residues that NisC stereospecifically couples to the dehydrated amino acids. Here, we demonstrate that increasing the number of negatively charged residues in the core peptide part of precursor nisin, which are absent in wild-type nisin, does not abolish binding of precursor nisin to the modification enzymes NisB and NisC, but dramatically decreases the antimicrobial potency of these nisin mutants. An unnatural precursor nisin variant lacking all serines and threonines in the core peptide part and an unnatural precursor nisin variant lacking all cysteines in the core peptide part still bind the nisin modification enzymes NisB and NisC, suggesting that these residues are not essential for direct interactions with the nisin modification enzymes NisB and NisC. These results are important for lantibiotic engineering studies.

  2. 改性糠醛渣对废水中镍离子的吸附性能%Adsorption Performance of Modified Furfural Residue for Nickel Ion

    Institute of Scientific and Technical Information of China (English)

    任广军; 邢琦; 王晓朋

    2013-01-01

    Adsorption properties of modified furfural residue for nickel ion was investigted.The results showed that the adsorption capacity of modified furfural residue for nickel ion in wastewater was reached to balance at 30 min.In the wastewater with volume of 50 mL and nickel concentration of 20 mg/L,the adsorption capacity of 0.02 g modified furfural residue for nickel ions was about 19 mg/g.Relationship between the equilibrium adsorption capacity and concentration was accorded with the Freundlich and Langmuir isothermal adsorption equation.%研究了改性糠醛渣对镍离子的吸附性能.结果表明,改性糠醛渣对废水中镍离子的吸附在30 min达到平衡;在50 mL含20mg/L的镍离子废水中,加入改性糠醛渣0.02g时,对废水中镍离子的吸附量达到19mg/g左右;平衡吸附量与平衡浓度之间的关系符合Freundlich和Langmuir等温吸附方程所描述的规律.

  3. Digestible lysine requirements of broilers

    Directory of Open Access Journals (Sweden)

    LEP Bernal

    2014-03-01

    Full Text Available Modern broilers have been submitted to continuous genetic improvement, and therefore, their nutritional requirements must be constantly updated to ensure their performance. Two experiments were carried out to evaluate different digestible lysine levels for starter (1021 days and grower (22-35 days phases. The experiments were carried out with male and female Cobb 500 broilers, distributed according to a randomized block experimental design in a 5x2 factorial arrangement (5 increasing digestible lysine levels x 2 sexes, totaling 10 treatments, with 8 replicates of 22 and 20 birds during the starter and grower phase, respectively. Digestible lysine levels of 1.06, 1.12, 1.18, 1.24, and 1.30 were used in the starter diets (10-21 days and 0.9, 0.98, 1.04, 1.10, and 1.16% in the grower diets (22-35 days. Based on the statistical analyses of the evaluated performance parameters, digestible lysine requirements for maximum performance were determined as 1.22% for males and 1.24% for females in the starter phase, and 1.16% for both sexes in the grower phase. Carcass and performance results indicate that digestible lysine requirements vary with sex and evaluated production parameter. Considering the most relevant broiler production parameters, in 22- to 35-d-old males, digestible lysine requirement for breast meat yield (1.16% was higher than those for feed conversion ratio (1.07% and weight gain (1.05%.

  4. DNA Damage-Induced Acetylation of Lysine 3016 of ATM Activates ATM Kinase Activity▿ †

    OpenAIRE

    Sun, Yingli; Xu, Ye; Roy, Kanaklata; Price, Brendan D.

    2007-01-01

    The ATM protein kinase is essential for cells to repair and survive genotoxic events. The activation of ATM's kinase activity involves acetylation of ATM by the Tip60 histone acetyltransferase. In this study, systematic mutagenesis of lysine residues was used to identify regulatory ATM acetylation sites. The results identify a single acetylation site at lysine 3016, which is located in the highly conserved C-terminal FATC domain adjacent to the kinase domain. Antibodies specific for acetyl-ly...

  5. Extensive Lysine Methylation in Hyperthermophilic Crenarchaea: Potential Implications for Protein Stability and Recombinant Enzymes

    Directory of Open Access Journals (Sweden)

    Catherine H. Botting

    2010-01-01

    Full Text Available In eukarya and bacteria, lysine methylation is relatively rare and is catalysed by sequence-specific lysine methyltransferases that typically have only a single-protein target. Using RNA polymerase purified from the thermophilic crenarchaeum Sulfolobus solfataricus, we identified 21 methyllysines distributed across 9 subunits of the enzyme. The modified lysines were predominantly in α-helices and showed no conserved sequence context. A limited survey of the Thermoproteus tenax proteome revealed widespread modification with 52 methyllysines in 30 different proteins. These observations suggest the presence of an unusual lysine methyltransferase with relaxed specificity in the crenarchaea. Since lysine methylation is known to enhance protein thermostability, this may be an adaptation to a thermophilic lifestyle. The implications of this modification for studies and applications of recombinant crenarchaeal enzymes are discussed.

  6. Systematic Analysis of the Functions of Lysine Acetylation in the Regulation of Tat Activity.

    Directory of Open Access Journals (Sweden)

    Minghao He

    Full Text Available The Tat protein of HIV-1 has several well-known properties, such as nucleocytoplasmic trafficking, transactivation of transcription, interaction with tubulin, regulation of mitotic progression, and induction of apoptosis. Previous studies have identified a couple of lysine residues in Tat that are essential for its functions. In order to analyze the functions of all the lysine residues in Tat, we mutated them individually to alanine, glutamine, and arginine. Through systematic analysis of the lysine mutants, we discovered several previously unidentified characteristics of Tat. We found that lysine acetylation could modulate the subcellular localization of Tat, in addition to the regulation of its transactivation activity. Our data also revealed that lysine mutations had distinct effects on microtubule assembly and Tat binding to bromodomain proteins. By correlation analysis, we further found that the effects of Tat on apoptosis and mitotic progression were not entirely attributed to its effect on microtubule assembly. Our findings suggest that Tat may regulate diverse cellular activities through binding to different proteins and that the acetylation of distinct lysine residues in Tat may modulate its interaction with various partners.

  7. Apolipoprotein A-I lysine modification: effects on helical content, lipid binding and cholesterol acceptor activity.

    Science.gov (United States)

    Brubaker, Gregory; Peng, Dao-Quan; Somerlot, Benjamin; Abdollahian, Davood J; Smith, Jonathan D

    2006-01-01

    We examined the role of the positively charged lysine residues in apoAI by chemical modification. Lysine modification by reductive methylation did not alter apoAI's net charge, secondary or tertiary structure as observed by circular dichroism and trytophan fluorescence, respectively, or have much impact on lipid binding or ABCA1-dependent cholesterol acceptor activity. Acetylation of lysine residues lowered the isoelectric point of apoAI, altered its secondary and tertiary structure, and led to a 40% decrease in cholesterol acceptor activity, while maintaining 93% of its lipid binding activity. Exhaustive lysine acetoacetylation lowered apoAI's isoelectric point, profoundly disrupted its secondary and tertiary structure, and led to 90% and 82% reductions in cholesterol acceptor and lipid binding activities, respectively. The dose-dependent acetoacetylation of an increasing proportion of apoAI lysine residues demonstrated that cholesterol acceptor activity was more sensitive to this modification than lipid binding activity, suggesting that apoAI lysine positive charges play an important role in ABCA1 mediated lipid efflux beyond the role needed to maintain alpha-helical content and lipid binding activity.

  8. Improvement of retroviral vectors by coating with poly(ethylene glycol)-poly(L-lysine) block copolymer (PEG-PLL).

    Science.gov (United States)

    Katakura, Hiromichi; Harada, Atsushi; Kataoka, Kazunori; Furusho, Miki; Tanaka, Fumihiro; Wada, Hiromi; Ikenaka, Kazuhiro

    2004-04-01

    Although some cationic reagents, such as polybrene, improve gene transduction in vitro, their use in vivo is prohibited due to their toxicity to the exposed cells. This paper demonstrates that a new cationic reagent, poly(ethylene glycol)-poly(L-lysine) block copolymer (PEG-PLL), improves gene transduction with retroviral vectors without increasing cell toxicity. A retroviral vector derived from the Moloney leukemia virus, containing the lacZ gene, was modified with PEG-PLL prior to transduction into NIH3T3, Lewis lung carcinoma, and primary cultured mouse brain cells. LacZ transduction efficacy was evaluated by counting the number of X-Gal-positive cells. We have demonstrated that PEG-PLL is able to stably modify the viral particle surface due to the affinity of the PEG moiety to the biomembrane, and neutralizes negative charges by the cationic nature of the poly-lysine residue. Thus, PEG-PLL increased the gene transduction efficiency and minimized cell toxicity because free PEG-PLL was removable by centrifugation. We have shown that PEG-PLL increased the viral gene transduction efficiency 3- to 7-fold with NIH3T3 or Lewis lung carcinoma cell lines without increasing cytotoxicity. It improved retroviral gene transduction efficacy even against labile cells, such as primary cultured brain cells. PEG-PLL is a novel reagent that improves retroviral gene transduction efficacy without increasing cytotoxicity. Copyright 2004 John Wiley & Sons, Ltd.

  9. Benzimidazole Carbamate Residues in Milk: Detection by SPR Biosensor; using a Modified QuEChERS Method for Extraction

    Science.gov (United States)

    A surface plasmon resonance (SPR) biosensor screening assay was developed and validated to detect 11 benzimidazole carbamate (BZT) veterinary drug residues in milk. The polyclonal antibody used was raised in sheep against a methyl 5 (6)-[(carboxypentyl)-thio]-2-benzimidazole carbamate protein conjug...

  10. Lysine-Grafted MCM-41 Silica as an Antibacterial Biomaterial

    Directory of Open Access Journals (Sweden)

    María F. Villegas

    2017-09-01

    Full Text Available This paper proposes a facile strategy for the zwitterionization of bioceramics that is based on the direct incorporation of l-lysine amino acid via the ε-amino group onto mesoporous MCM-41 materials. Fourier transform infrared (FTIR studies of lysine-grafted MCM-41 (MCM-LYS simultaneously showed bands at 3080 and 1540 cm−1 and bands at 1625 and 1415 cm−1 corresponding to -NH3+/COO− pairs, which demonstrate the incorporation of the amino acid on the material surface keeping its zwitterionic character. Both elemental and thermogravimetric analyses showed that the amount of grafted lysine was 8 wt. % based on the bioceramic total weight. Moreover, MCM-LYS exhibited a reduction of adhesion of S. aureus and E. coli bacteria in 33% and 50%, respectively at physiological pH, as compared with pristine MCM-41. Biofilm studies onto surfaces showed that lysine functionalization elicited a reduction of the area covered by S. aureus biofilm from 42% to only 5% (88%. This research shows a simple and effective approach to chemically modify bioceramics using single amino acids that provides zwitterionic functionality, which is useful to develop new biomaterials that are able to resist bacterial adhesion.

  11. Use of Oxalic-Acid-Modified Stellerite for Improving the Filter Capability of PM2.5 of Paper Composed of Bamboo Residues

    Directory of Open Access Journals (Sweden)

    Hua Chen

    2016-01-01

    Full Text Available In this study, pulping conditions for kraft pulping of bamboo residues were investigated, predominantly focusing on cooking temperature and time during pulping. Oxalic acid and cationic starch were used for the modification of natural stellerite, and the use of modified stellerite for preparing filter paper for PM2.5 filtration was investigated. The optimal pulping technology of bamboo residues was established based on the following experimental parameters: liquor ratio of 1 : 5.5, cooking temperature of 160°C, and a holding time of 2 h. Modification by oxalic acid resulted in the promotion of pore formation at the stellerite surfaces and induced the microscopic changes. Nevertheless, paper strength remained practically unchanged after the addition of fillers, indicating that the cationic starch preblend method is a promising technique for papermaking because it enhances the strength properties of paper. With the variation in the addition of modified stellerite from 3 to 15%, while simultaneously maintaining the basis weight constant at 60 gm−2, the filtration efficiency of paper sheets first increased and then decreased later; thus the optimum stellerite content was found to be 9%. Filtration efficiency was suggested to be affected by gas flowing velocity.

  12. Available lysine in canned fish

    OpenAIRE

    Rao, D. Ramananda; Gadre, Ujjwala V.

    1984-01-01

    Otolithus argenteus was canned in brine by heat processing at two different steam pressures either at 0.70 kg/cm super(2) or 1.05 kg/cm super(2) for 25 minutes. The nutritive value of canned fish as evaluated by the total nitrogen and available lysine did not alter much either during heat processing or during storage over a period of nine months at 28 degree plus or minus 5 degree C.

  13. BIOLOGICAL ADHESIVES. Adaptive synergy between catechol and lysine promotes wet adhesion by surface salt displacement.

    Science.gov (United States)

    Maier, Greg P; Rapp, Michael V; Waite, J Herbert; Israelachvili, Jacob N; Butler, Alison

    2015-08-01

    In physiological fluids and seawater, adhesion of synthetic polymers to solid surfaces is severely limited by high salt, pH, and hydration, yet these conditions have not deterred the evolution of effective adhesion by mussels. Mussel foot proteins provide insights about adhesive adaptations: Notably, the abundance and proximity of catecholic Dopa (3,4-dihydroxyphenylalanine) and lysine residues hint at a synergistic interplay in adhesion. Certain siderophores—bacterial iron chelators—consist of paired catechol and lysine functionalities, thereby providing a convenient experimental platform to explore molecular synergies in bioadhesion. These siderophores and synthetic analogs exhibit robust adhesion energies (E(ad) ≥-15 millijoules per square meter) to mica in saline pH 3.5 to 7.5 and resist oxidation. The adjacent catechol-lysine placement provides a "one-two punch," whereby lysine evicts hydrated cations from the mineral surface, allowing catechol binding to underlying oxides.

  14. PLMD: An updated data resource of protein lysine modifications.

    Science.gov (United States)

    Xu, Haodong; Zhou, Jiaqi; Lin, Shaofeng; Deng, Wankun; Zhang, Ying; Xue, Yu

    2017-05-20

    Post-translational modifications (PTMs) occurring at protein lysine residues, or protein lysine modifications (PLMs), play critical roles in regulating biological processes. Due to the explosive expansion of the amount of PLM substrates and the discovery of novel PLM types, here we greatly updated our previous studies, and presented a much more integrative resource of protein lysine modification database (PLMD). In PLMD, we totally collected and integrated 284,780 modification events in 53,501 proteins across 176 eukaryotes and prokaryotes for up to 20 types of PLMs, including ubiquitination, acetylation, sumoylation, methylation, succinylation, malonylation, glutarylation, glycation, formylation, hydroxylation, butyrylation, propionylation, crotonylation, pupylation, neddylation, 2-hydroxyisobutyrylation, phosphoglycerylation, carboxylation, lipoylation and biotinylation. Using the data set, a motif-based analysis was performed for each PLM type, and the results demonstrated that different PLM types preferentially recognize distinct sequence motifs for the modifications. Moreover, various PLMs synergistically orchestrate specific cellular biological processes by mutual crosstalks with each other, and we totally found 65,297 PLM events involved in 90 types of PLM co-occurrences on the same lysine residues. Finally, various options were provided for accessing the data, while original references and other annotations were also present for each PLM substrate. Taken together, we anticipated the PLMD database can serve as a useful resource for further researches of PLMs. PLMD 3.0 was implemented in PHP + MySQL and freely available at http://plmd.biocuckoo.org. Copyright © 2017 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Ltd. All rights reserved.

  15. Detección daño estructural empleando el vector de fuerza residual modificado y el algoritmo Simulated Annealing (SA Damage Detection Using the Modified Residual Force Vector and the Simulated Annealing Algorithm (SA

    Directory of Open Access Journals (Sweden)

    Óscar Begambre

    2010-01-01

    Full Text Available En este trabajo, el algoritmo Simulated Annealing (SA es empleado para solucionar el problema inverso de detección de daño en vigas usando información modal contaminada con ruido. La formulación de la función objetivo para el procedimiento de optimización, basado en el SA, está fundamentada en el método de la fuerza residual modificada. El desempeño del SA empleado en este estudio superó el de un algoritmo genético (AG en dos funciones de prueba reportadas en la literatura internacional. El procedimiento de evaluación de integridad aquí propuesto se confirmó y validó numéricamente empleando la teoría de vigas de Euler-Bernoulli y un Modelo de Elementos Finitos (MEF de vigas en voladizo y apoyadas libremente.In this research, the Simulated Annealing Algorithm (SA is employed to solve damage detection problems in beam type structures using noisy polluted modal data. The formulation of the objective function for the SA optimization procedure is based on the modified residual force method. The SA used in this research performs better than the Genetic Algorithm (GA in two difficult benchmark functions. The proposed structural damage-identification scheme is confirmed and assessed using a Finite Element Model (FEM of cantilever and a free-free Euler-Bernoulli beam model

  16. Entamoeba histolytica: protein arginine transferase 1a methylates arginine residues and potentially modify the H4 histone.

    Science.gov (United States)

    Borbolla-Vázquez, Jessica; Orozco, Esther; Betanzos, Abigail; Rodríguez, Mario A

    2015-04-10

    In eukaryotes, histone arginine methylation associates with both active and repressed chromatin states depending on the residues involved and the status of methylation. Even when the amino-terminus of Entamoeba histolytica histones diverge from metazoan sequences, these regions contain arginine residues that are potential targets for methylation. However, histone arginine methylation as well as the activity of arginine methyltransferases (PRMTs) has not been studied in this parasite. The aim of this work was to examine the dimethylation of arginine 3 of H4 histone (H4R3me2) and to identify the parasite PRMT that could be responsible for this modification (EhPRMT1). To examine the presence of H4R3me2 in E histolytica, we performed Western blot and immunofluorescence assays on trophozoites using an antibody against this epigenetic mark. To recognize the PRMT1 enzyme of this parasite that possibly perform that modification, we first performed a phylogenetic analysis of E. histolytica and human PRMTs. RT-PCR assays were carried out to analyze the expression of the putative PRMT1 genes. One of these genes was cloned and expressed in Escherichia coli. The recombinant protein was tested by its recognition by an antibody against human PRMT1 and in its ability to form homodimers and to methylate commercial histones. The arginine 3 of human H4, which is subjected to post translational methylation, was aligned with the arginine 8 of E. histolytica H4, suggesting that this residue could be methylated. The recognition of an 18 kDa nuclear protein of E. histolytica by an antibody against H4R3me2 confirmed this assumption. We found that this parasite expresses three phylogenetic and structural proteins related to PRMT1. Antibodies against the human PRMT1 detected E. histolytica proteins in cytoplasm and nuclei and recognized a recombinant PRMT1 of this parasite. The recombinant protein was able to form homodimers and homotetramers and displayed methyltransferase activity on

  17. Identification and characterization of lysine-methylated sites on histones and non-histone proteins.

    Science.gov (United States)

    Lee, Tzong-Yi; Chang, Cheng-Wei; Lu, Cheng-Tzung; Cheng, Tzu-Hsiu; Chang, Tzu-Hao

    2014-06-01

    Protein methylation is a kind of post-translational modification (PTM), and typically takes place on lysine and arginine amino acid residues. Protein methylation is involved in many important biological processes, and most recent studies focused on lysine methylation of histones due to its critical roles in regulating transcriptional repression and activation. Histones possess highly conserved sequences and are homologous in most species. However, there is much less sequence conservation among non-histone proteins. Therefore, mechanisms for identifying lysine-methylated sites may greatly differ between histones and non-histone proteins. Nevertheless, this point of view was not considered in previous studies. Here we constructed two support vector machine (SVM) models by using lysine-methylated data from histones and non-histone proteins for predictions of lysine-methylated sites. Numerous features, such as the amino acid composition (AAC) and accessible surface area (ASA), were used in the SVM models, and the predictive performance was evaluated using five-fold cross-validations. For histones, the predictive sensitivity was 85.62% and specificity was 80.32%. For non-histone proteins, the predictive sensitivity was 69.1% and specificity was 88.72%. Results showed that our model significantly improved the predictive accuracy of histones compared to previous approaches. In addition, features of the flanking region of lysine-methylated sites on histones and non-histone proteins were also characterized and are discussed. A gene ontology functional analysis of lysine-methylated proteins and correlations of lysine-methylated sites with other PTMs in histones were also analyzed in detail. Finally, a web server, MethyK, was constructed to identify lysine-methylated sites. MethK now is available at http://csb.cse.yzu.edu.tw/MethK/.

  18. A novel amperometric biosensor based on a co-crosslinked L-lysine-α-oxidase/overoxidized polypyrrole bilayer for the highly selective determination of L-lysine.

    Science.gov (United States)

    Guerrieri, Antonio; Ciriello, Rosanna; Cataldi, Tommaso R I

    2013-09-17

    An amperometric biosensor for the determination of L-lysine based on L-lysine-α-oxidase immobilized by co-crosslinking on a platinum electrode previously modified by an overoxidized polypyrrole film is described. The optimization of experimental parameters, such as pH and flow rate, permitted to minimize significantly substrate interferences even using a low specific, commercial enzyme. The relevant biases introduced in the measurement of lysine were just about 1% for L-arginine, L-histidine and L-ornithine, roughly 4% for L-phenylalanine and L-tyrosine. The developed approach allowed linear lysine responses from 0.02 mM up to 2 mM with a sensitivity of 41 nA/(mM × mm(2)) and a detection limit of 4 μM (S/N=3). No appreciable loss in lysine sensitivity was observed up to about 40 days. Allowing polypyrrole layer to remove interference from electroactive compounds, the present method revealed suitable to detect L-lysine in a pharmaceutical and cheese sample, showing a good agreement with the expected values. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Measurement of the Residual Gases O2 and CO2 in Meat Products Packed in Modified Atmosphere

    Directory of Open Access Journals (Sweden)

    Jozef Čapla

    2013-02-01

    Full Text Available Nowadays, consumers have increased demand for quality and food safety and also rising demand for natural foods without chemical additives. There are many ways to presserve freshness of these products, one of them is modified atmosphere packaging, which can mean elimination and/or replacement surrounding the product before closing it in package with a mixture of gases other than the original ambient air atmosphere. for replacement of atmosphere are generally used three types of gases such as carbon dioxide, oxygen and nitrogen. this type of packaging is often used for meat and meat products, which belongs to foods that are under normal conditions perishable and for increasing the shelf life of meat products are also used various other preservation methods or their combinations. Packaging of meat and meat products in modified atmosphere is usually made with a high content of carbon dioxide, which has good bacteriostatic and fungistatic effect and is also an effective mean for increasing the shelf life of packaged products during storage and sale.

  20. The SUVR4 histone lysine methyltransferase binds ubiquitin and converts H3K9me1 to H3K9me3 on transposon chromatin in Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Silje V Veiseth

    2011-03-01

    Full Text Available Chromatin structure and gene expression are regulated by posttranslational modifications (PTMs on the N-terminal tails of histones. Mono-, di-, or trimethylation of lysine residues by histone lysine methyltransferases (HKMTases can have activating or repressive functions depending on the position and context of the modified lysine. In Arabidopsis, trimethylation of lysine 9 on histone H3 (H3K9me3 is mainly associated with euchromatin and transcribed genes, although low levels of this mark are also detected at transposons and repeat sequences. Besides the evolutionarily conserved SET domain which is responsible for enzyme activity, most HKMTases also contain additional domains which enable them to respond to other PTMs or cellular signals. Here we show that the N-terminal WIYLD domain of the Arabidopsis SUVR4 HKMTase binds ubiquitin and that the SUVR4 product specificity shifts from di- to trimethylation in the presence of free ubiquitin, enabling conversion of H3K9me1 to H3K9me3 in vitro. Chromatin immunoprecipitation and immunocytological analysis showed that SUVR4 in vivo specifically converts H3K9me1 to H3K9me3 at transposons and pseudogenes and has a locus-specific repressive effect on the expression of such elements. Bisulfite sequencing indicates that this repression involves both DNA methylation-dependent and -independent mechanisms. Transcribed genes with high endogenous levels of H3K4me3, H3K9me3, and H2Bub1, but low H3K9me1, are generally unaffected by SUVR4 activity. Our results imply that SUVR4 is involved in the epigenetic defense mechanism by trimethylating H3K9 to suppress potentially harmful transposon activity.

  1. Hemoglobin Labeled by Radioactive Lysine

    Science.gov (United States)

    Bale, W. F.; Yuile, C. L.; DeLaVergne, L.; Miller, L. L.; Whipple, G. H.

    1949-12-08

    This paper reports on the utilization of tagged epsilon carbon of DL-lysine by a dog both anemic and hypoproteinemic due to repeated bleeding plus a diet low in protein. The experiment extended over period of 234 days, a time sufficient to indicate an erythrocyte life span of at least 115 days based upon the rate of replacement of labeled red cell proteins. The proteins of broken down red cells seem not to be used with any great preference for the synthesis of new hemoglobin.

  2. Interferon α: the salvage therapy for patients with unsatisfactory response to minimal residual disease-directed modified donor lymphocyte infusion

    Institute of Scientific and Technical Information of China (English)

    Mo Xiaodong; Zhao Xiangyu; Xu Lanping; Liu Daihong; Zhang Xiaohui; Chen Huan; Wang Yu

    2014-01-01

    Background Minimal residual disease (MRD)-directedmodified donor lymphocyte infusion (mDLI) is used to treat relapse after hematopoietic stem cell transplantation (HSCT).For patients who experience an unsatisfactory response tomDLI,relapse is usually inevitable.Therefore,we sought to evaluate the efficacy ofinterferon α therapy in these patients.Methods Regular MRD monitoring was carried out after the HSCT.The patients who were MRD-positive underwent mDLI.Patients with an unsatisfactory response to mDLI received interferon α therapy (3 million units,twice weekly) with regular monitoring of MRD.To ensure the immunomodulatory effects of interferon α,immunosuppressant treatment would be stopped before interferon α treatment.Results Five patients with an unsatisfactory response to mDLI treatment received interferon α (3 had t(8;21) chromosomal translocation acute myeloid leukemia,and 2 had common acute leukemia).They had significantly reduced or resolved MRD.Four patients developed chronic graft-versus-host disease.Two of the 5 patients reported transient fevers,and no significant bone marrow suppression was observed.All of them were in continuous complete remission after interferon α treatment.The median survival time was 469 days (range 368-948 days).Conclusions In patients with an unsatisfactory response to MRD-directed mDLI,interferon α may directly or indirectly induce the graft-versus-leukemia effect to improve mDLI efficacy and clear MRD.

  3. Autoacetylation of the MYST lysine acetyltransferase MOF protein.

    Science.gov (United States)

    Yang, Chao; Wu, Jiang; Sinha, Sarmistha H; Neveu, John M; Zheng, Yujun George

    2012-10-12

    The MYST family of histone acetyltransferases (HATs) plays critical roles in diverse cellular processes, such as the epigenetic regulation of gene expression. Lysine autoacetylation of the MYST HATs has recently received considerable attention. Nonetheless, the mechanism and function of the autoacetylation process are not well defined. To better understand the biochemical mechanism of MYST autoacetylation and the impact of autoacetylation on the cognate histone acetylation, we carried out detailed analyses of males-absent-on-the-first (MOF), a key member of the MYST family. A number of mutant MOF proteins were produced with point mutations at several key residues near the active site of the enzyme. Autoradiography and immunoblotting data showed that mutation of these residues affects the autoacetylation activity and HAT activity of MOF by various degrees demonstrating that MOF activity is highly sensitive to the chemical changes in those residues. We produced MOF protein in the deacetylated form by using a nonspecific lysine deacetylase. Interestingly, both the autoacetylation activity and the histone acetylation activity of the deacetylated MOF were found to be very close to that of wild-type MOF, suggesting that autoacetylation of MOF only marginally modulates the enzymatic activity. Also, we found that the autoacetylation rates of MOF and deacetylated MOF were much slower than the cognate substrate acetylation. Thus, autoacetylation does not seem to contribute to the intrinsic enzymatic activity in a significant manner. These data provide new insights into the mechanism and function of MYST HAT autoacetylation.

  4. Autoacetylation of the MYST Lysine Acetyltransferase MOF Protein*

    Science.gov (United States)

    Yang, Chao; Wu, Jiang; Sinha, Sarmistha H.; Neveu, John M.; Zheng, Yujun George

    2012-01-01

    The MYST family of histone acetyltransferases (HATs) plays critical roles in diverse cellular processes, such as the epigenetic regulation of gene expression. Lysine autoacetylation of the MYST HATs has recently received considerable attention. Nonetheless, the mechanism and function of the autoacetylation process are not well defined. To better understand the biochemical mechanism of MYST autoacetylation and the impact of autoacetylation on the cognate histone acetylation, we carried out detailed analyses of males-absent-on-the-first (MOF), a key member of the MYST family. A number of mutant MOF proteins were produced with point mutations at several key residues near the active site of the enzyme. Autoradiography and immunoblotting data showed that mutation of these residues affects the autoacetylation activity and HAT activity of MOF by various degrees demonstrating that MOF activity is highly sensitive to the chemical changes in those residues. We produced MOF protein in the deacetylated form by using a nonspecific lysine deacetylase. Interestingly, both the autoacetylation activity and the histone acetylation activity of the deacetylated MOF were found to be very close to that of wild-type MOF, suggesting that autoacetylation of MOF only marginally modulates the enzymatic activity. Also, we found that the autoacetylation rates of MOF and deacetylated MOF were much slower than the cognate substrate acetylation. Thus, autoacetylation does not seem to contribute to the intrinsic enzymatic activity in a significant manner. These data provide new insights into the mechanism and function of MYST HAT autoacetylation. PMID:22918831

  5. Evaluation of the interfacial shear strength and residual stress of TiAlN coating on ZIRLO™ fuel cladding using a modified shear-lag model approach

    Science.gov (United States)

    Liu, Y.; Bhamji, I.; Withers, P. J.; Wolfe, D. E.; Motta, A. T.; Preuss, M.

    2015-11-01

    This paper investigates the residual stresses and interfacial shear strength of a TiAlN coating on Zr-Nb-Sn-Fe alloy (ZIRLO™) substrate designed to improve corrosion resistance of fuel cladding used in water-cooled nuclear reactors, both during normal and exceptional conditions, e.g. a loss of coolant event (LOCA). The distribution and maximum value of the interfacial shear strength has been estimated using a modified shear-lag model. The parameters critical to this analysis were determined experimentally. From these input parameters the interfacial shear strength between the TiAlN coating and ZIRLO™ substrate was inferred to be around 120 MPa. It is worth noting that the apparent strength of the coating is high (∼3.4 GPa). However, this is predominantly due to the large compressive residuals stress (3 GPa in compression), which must be overcome for the coating to fail in tension, which happens at a load just 150 MPa in excess of this.

  6. Separating nano graphene oxide from the residual strong-acid filtrate of the modified Hummers method with alkaline solution

    Science.gov (United States)

    Hu, Xuebing; Yu, Yun; Wang, Yongqing; Zhou, Jianer; Song, Lixin

    2015-02-01

    In the modified Hummers method for preparing graphene oxide, the yellow slurry can be obtained. After filtering through a quantitative filter paper, the strong-acid filtrate containing the unprecipitated nano graphene oxide was gained. The corresponding filtrate was added gradually with an alkaline (NaOH or KOH) solution at room temperature. The unprecipitated nano graphene oxide could undergo fast aggregation when the pH value of the filtrate was about 1.7 and formed the stable floccules. X-ray diffraction analysis shows the dominant peak of the floccules is about 11°, which accords to the peak of graphene oxide. Spectra of X-ray photoelectron spectroscopy confirm the presence in the floccules of an abundance of oxygen functional groups and the purified graphene oxide floccules can be obtained. Atomic force microscopy measurement shows the graphene oxide floccules consists of sheet-like objects, mostly containing only a few layers (about 5 layers). Zeta potential analysis demonstrates the surface charge of the graphene oxide is pH-sensitive and its isoelectric point is ∼1.7. The flocculation mechanism of graphene oxide ascribes to the acid-base interaction with the surface functional groups of the carbon layers.

  7. PENILAIAN PENGARUH PENAMBAHAN LYSINE PADA NASI

    Directory of Open Access Journals (Sweden)

    Ignatius Tarwotjo

    2012-11-01

    Full Text Available Pengaruh penambahan lysine pada mutu protein nasi dilakukan pada tikus putih dengan mengukur Protein Efficiency Ratio. Nasi dan Nasi dengan sayur beserta laukpauk, seperti dikonsumsi oleh kebanyakan keluarga di Indonesia, yang berasnya lebih dulu ditambahi butiran premix berisi lysine, thiamine dan riboflavin ternaya menghasilkan Protein Efficiency Ratio lebih tinggi dari pada yang tidak ditambahi.

  8. The CF-modifying gene EHF promotes p.Phe508del-CFTR residual function by altering protein glycosylation and trafficking in epithelial cells.

    Science.gov (United States)

    Stanke, Frauke; van Barneveld, Andrea; Hedtfeld, Silke; Wölfl, Stefan; Becker, Tim; Tümmler, Burkhard

    2014-05-01

    The three-base-pair deletion c.1521_1523delCTT (p.Phe508del, F508del) in the cystic fibrosis transmembrane conductance regulator (CFTR) is the most frequent disease-causing lesion in cystic fibrosis (CF). The CFTR gene encodes a chloride and bicarbonate channel at the apical membrane of epithelial cells. Altered ion transport of CFTR-expressing epithelia can be used to differentiate manifestations of the so-called CF basic defect. Recently, an 11p13 region has been described as a CF modifier by the North American CF Genetic Modifier Study Consortium. Selecting the epithelial-specific transcription factor EHF (ets homologous factor) as the likely candidate gene on 11p13, we have genotyped two intragenic microsatellites in EHF to replicate the 11p13 finding in the patient cohort of the European CF Twin and Sibling Study. We could observe an association of rare EHF haplotypes among homozygotes for c.1521_1523delCTT in CFTR, which exhibit a CF-untypical manifestation of the CF basic defect such as CFTR-mediated residual chloride secretion and low response to amiloride. We have reviewed transcriptome data obtained from intestinal epithelial samples of homozygotes for c.1521_1523delCTT in CFTR, which were stratified for their EHF genetic background. Transcripts that were upregulated among homozygotes for c.1521_1523delCTT in CFTR, who carry two rare EHF alleles, were enriched for genes that alter protein glycosylation and trafficking, both mechanisms being pivotal for the effective targeting of fully functional p.Phe508del-CFTR to the apical membrane of epithelial cells. We conclude that EHF modifies the CF phenotype by altering capabilities of the epithelial cell to correctly process the folding and trafficking of mutant p.Phe508del-CFTR.

  9. Engineering a Lysine-ON Riboswitch for Metabolic Control of Lysine Production in Corynebacterium glutamicum.

    Science.gov (United States)

    Zhou, Li-Bang; Zeng, An-Ping

    2015-12-18

    Riboswitches are natural RNA elements that regulate gene expression by binding a ligand. Here, we demonstrate the possibility of altering a natural lysine-OFF riboswitch from Eschericia coli (ECRS) to a synthetic lysine-ON riboswitch and using it for metabolic control. To this end, a lysine-ON riboswitch library was constructed using tetA-based dual genetic selection. After screening the library, the functionality of the selected lysine-ON riboswitches was examined using a report gene, lacZ. Selected lysine-ON riboswitches were introduced into the lysE gene (encoding a lysine transport protein) of Corynebacterium glutamicum and used to achieve dynamic control of lysine transport in a recombinant lysine-producing strain, C. glutamicum LPECRS, which bears a deregulated aspartokinase and a lysine-OFF riboswitch for dynamic control of the enzyme citrate synthase. Batch fermentation results of the strains showed that the C. glutamicum LPECRS strain with an additional lysine-ON riboswitch for the control of lysE achieved a 21% increase in the yield of lysine compared to that of the C. glutamicum LPECRS strain and even a 89% increase in yield compared to that of the strain with deregulated aspartokinase. This work provides a useful approach to generate lysine-ON riboswitches for C. glutamicum metabolic engineering and demonstrates for the first time a synergetic effect of lysine-ON and -OFF riboswitches for improving lysine production in this industrially important microorganism. The approach can be used to dynamically control other genes and can be applied to other microorganisms.

  10. Jarid2 binds mono-ubiquitylated H2A lysine 119 to mediate crosstalk between Polycomb complexes PRC1 and PRC2

    DEFF Research Database (Denmark)

    Cooper, Sarah; Grijzenhout, Anne; Underwood, Elizabeth

    2016-01-01

    The Polycomb repressive complexes PRC1 and PRC2 play a central role in developmental gene regulation in multicellular organisms. PRC1 and PRC2 modify chromatin by catalysing histone H2A lysine 119 ubiquitylation (H2AK119u1), and H3 lysine 27 methylation (H3K27me3), respectively. Reciprocal...

  11. Site-Specific Conjugation to Native and Engineered Lysines in Human Immunoglobulins by Microbial Transglutaminase.

    Science.gov (United States)

    Spidel, Jared L; Vaessen, Benjamin; Albone, Earl F; Cheng, Xin; Verdi, Arielle; Kline, J Bradford

    2017-09-20

    The use of microbial transglutaminase (MTG) to produce site-specific antibody-drug conjugates (ADCs) has thus far focused on the transamidation of engineered acyl donor glutamine residues in an antibody based on the hypothesis that the lower specificity of MTG for acyl acceptor lysines may result in the transamidation of multiple native lysine residues, thereby yielding heterogeneous products. We investigated the utilization of native IgG lysines as acyl acceptor sites for glutamine-based acyl donor substrates. Of the approximately 80 lysines in multiple recombinant IgG monoclonal antibodies (mAbs), none were transamidated. Because recombinant mAbs lack the C-terminal Lys447 due to cleavage by carboxypeptidase B in the production cell host, we explored whether blocking the cleavage of Lys447 by the addition of a C-terminal amino acid could result in transamidation of Lys447 by a variety of acyl donor substrates. MTG efficiently transamidated Lys447 in the presence of any nonacidic, nonproline amino acid residue at position 448. Lysine scanning mutagenesis throughout the antibody further revealed several transamidation sites in both the heavy- and light-chain constant regions. Additionally, scanning mutagenesis of the hinge region in a Fab' fragment revealed sites of transamidation that were not reactive in the context of the full-length mAb. Here, we demonstrate the utility of single lysine substitutions and the C-terminal Lys447 for engineering efficient acyl acceptor sites suitable for site-specific conjugation to a range of glutamine-based acyl donor substrates.

  12. Increased responsiveness to dietary lysine deficiency of pectoralis major muscle protein turnover in broilers selected on breast development.

    Science.gov (United States)

    Tesseraud, S; Temim, S; Le Bihan-Duval, E; Chagneau, A M

    2001-04-01

    It has been previously established that growth and carcass qualities of chicks are modified by genotype and dietary amino acid supply. In this study, we evaluated the effects of lysine deficiency and genetic selection on muscle protein metabolism. Chicks originating from an experimental line selected for breast development (QL) and its control line (CL) were provided ad libitum access to isoenergetic diets containing 20% crude protein but differing in their lysine content (0.75 or 1.01%). Protein fractional synthesis rates (FSR) were measured in vivo in the pectoralis major and sartorius muscles of 3-wk-old chickens (flooding dose of [3H]phenylalanine). Fractional breakdown rates (FBR) were estimated as the difference between synthesis and deposition. Lysine deficiency reduced (P 0.14). In the pectoralis major muscle, chicks of both lines receiving an adequate lysine intake also exhibited similar protein turnover rates. However, in chicks fed the lysine-deficient (0.75% lysine) diet, FSR and Cs were higher in QL than in CL chicks (P < 0.05), and FBR tended (P = 0.07) to be higher in QL chicks. This increased protein turnover in the QL birds on the lysine-deficient diet suggests that the responsiveness of muscle protein metabolism to amino acid supply is modified by genetic selection for breast development.

  13. Acetyl-Phosphate Is a Critical Determinant of Lysine Acetylation in E. coli

    DEFF Research Database (Denmark)

    Weinert, Brian T; Iesmantavicius, Vytautas; Wagner, Sebastian A

    2013-01-01

    Lysine acetylation is a frequently occurring posttranslational modification in bacteria; however, little is known about its origin and regulation. Using the model bacterium Escherichia coli (E. coli), we found that most acetylation occurred at a low level and accumulated in growth-arrested cells...... acetylate lysine residues in vitro and that AcP levels are correlated with acetylation levels in vivo, suggesting that AcP may acetylate proteins nonenzymatically in cells. These results uncover a critical role for AcP in bacterial acetylation and indicate that most acetylation in E. coli occurs at a low...

  14. Modified Dwyer osteotomy with rotation and reinsertion of autograft bone wedge for residual heel deformity despite previous delayed subtalar joint arthrodesis after calcaneal fracture.

    Science.gov (United States)

    Boffeli, Troy J; Abben, Kyle W

    2014-01-01

    Calcaneal fracture patterns vary widely, and many factors determine the type and timing of the treatment rendered. Severe calcaneus fractures involving joint damage, loss of heel height, and varus deformity of the tuberosity are ideally treated with open reduction and internal fixation to repair the joint surface and re-establish anatomic structure. This is not always possible owing to delayed presentation, soft tissue compromise, unrelated injuries, unstable medical condition, or lack of expertise by the treating physician. We present the case of a patient who had residual forefoot and rearfoot deformity despite undergoing delayed subtalar joint arthrodesis at an outside hospital 10 years before for a calcaneal fracture that was initially treated nonoperatively. At 4 years of follow-up after modified Dwyer calcaneal osteotomy with rotation and reinsertion of the autograft bone wedge and Cotton midfoot osteotomy, the postoperative gait was relatively normal, other than the expected lack of hindfoot mobility. The lateral column pain was resolved. The patient remained highly satisfied with the outcome at long-term follow-up of 48 months, with improved heel alignment, lack of a wide stance gait, a functional medial column, and a relatively normal gait. This case demonstrates the value of periarticular calcaneal osteotomies without the need to revise the subtalar joint arthrodesis for this challenging clinical situation. Copyright © 2014 American College of Foot and Ankle Surgeons. Published by Elsevier Inc. All rights reserved.

  15. The synergistic effect on production of lignin-modifying enzymes through submerged co-cultivation of Phlebia radiata, Dichomitus squalens and Ceriporiopsis subvermispora using agricultural residues.

    Science.gov (United States)

    Dong, Ya-Chen; Wang, Wei; Hu, Zhong-Ce; Fu, Ming-Liang; Chen, Qi-He

    2012-06-01

    The lignin-modifying enzymes (LMEs) play an important role in decomposition of agricultural residues, which contain a certain amount of lignin. In this study, the production of LMEs by three co-cultivated combinations of Phlebia radiata, Dichomitus squalens and Ceriporiopsis subvermispora and the respective monocultures was comparatively investigated. Laccase and manganese peroxidases (MnP) were significantly promoted in the co-culture of P. radiata and D. squalens, and corncob was verified to be beneficial for laccase and MnP production. Moreover, laccase production by co-culture of P. radiata and D. squalens with high ratio of glucose to nitrogen was higher than low ratio under carbon- and nitrogen-meager conditions. New laccase isoenzymes measured by Native-PAGE were stimulated by co-cultured P. radiata with D. squalens or C. subvermispora, respectively, growing in the defined medium containing corncob, but the expression of laccase was greatly restrained by the co-culturing of D. squalens with C. subvermispora. This study showed that the synergistic and depressing effects of co-cultivation of P. radiata, D. squalens and C. subvermispora on LMEs were species specific.

  16. Reactive lysine content in commercially available pet foods

    NARCIS (Netherlands)

    Rooijen, van C.; Bosch, G.; Poel, van der A.F.B.; Wierenga, P.A.; Alexander, L.; Hendriks, W.H.

    2014-01-01

    The Maillard reaction can occur during processing of pet foods. During this reaction, the e-amino group of lysine reacts with reducing sugars to become unavailable for metabolism. The aim of the present study was to determine the reactive lysine (RL; the remaining available lysine) to total lysine (

  17. Oligo(L-lysine)-induced titanium dioxide: Effects of consecutive lysine on precipitation

    Science.gov (United States)

    Ahn, Sungjun; Park, Sangwoo; Lee, Sang-Yup

    2011-11-01

    Biomineralization of metal oxide utilizes biomolecular substances, such as peptides and proteins, to induce mineralization of metal precursors in a mild aqueous solution. In this study, we investigated biomineralization of an abiological substance, titanium dioxide (TiO 2), by oligo(L-lysine). Specifically, we systemically studied the influence of the number of consecutive lysine on TiO 2 precipitation. Oligo(L-lysine) was chosen as a homopeptide lysine source whose lysine quantity was adjusted. When oligo(L-lysine) contains more than three consecutive lysine, it induces notably fast precipitation, while single and dilysine do not readily form TiO 2 precipitates. Precipitation of TiO 2 was promoted with the length of oligo(L-lysine). The oligo(L-lysine) was associated with TiO 2 precipitate, which was confirmed by spectroscopic and thermogravitational analyses. The outcomes of this research provide a plausible rationale for explaining precipitation of the Ti precursor that is highly dependent on peptide sequences.

  18. A Novel Staphylococcus Podophage Encodes a Unique Lysin with Unusual Modular Design

    Science.gov (United States)

    Cater, Katie; Dandu, Vidya Sree; Bari, S. M. Nayeemul; Lackey, Kim; Everett, Gabriel F. K.

    2017-01-01

    ABSTRACT Drug-resistant staphylococci, particularly Staphylococcus aureus and Staphylococcus epidermidis, are leading causes of hospital-acquired infections. Bacteriophages and their peptidoglycan hydrolytic enzymes (lysins) are currently being explored as alternatives to conventional antibiotics; however, only a limited diversity of staphylococcal phages and their lysins has yet been characterized. Here, we describe a novel staphylococcal phage and its lysins. Bacteriophage Andhra is the first reported S. epidermidis phage belonging to the family Podoviridae. Andhra possesses an 18,546-nucleotide genome with 20 open reading frames. BLASTp searches revealed that gene product 10 (gp10) and gp14 harbor putative catalytic domains with predicted peptidase and amidase activities, characteristic functions of phage lysins. We purified these proteins and show that both Andhra_gp10 and Andhra_gp14 inhibit growth and degrade cell walls of diverse staphylococci, with Andhra_gp10 exhibiting more robust activity against the panel of cell wall substrates tested. Site-directed mutagenesis of its predicted catalytic residues abrogated the activity of Andhra_gp10, consistent with the presence of a catalytic CHAP domain on its C terminus. The active site location combined with the absence of an SH3b cell wall binding domain distinguishes Andhra_gp10 from the majority of staphylococcal lysins characterized to date. Importantly, close homologs of Andhra_gp10 are present in related staphylococcal podophages, and we propose that these constitute a new class of phage-encoded lysins. Altogether, our results reveal insights into the biology of a rare family of staphylococcal phages while adding to the arsenal of antimicrobials with potential for therapeutic use. IMPORTANCE The spread of antibiotic resistance among bacterial pathogens is inciting a global public health crisis. Drug-resistant Staphylococcus species, especially S. aureus and S. epidermidis, have emerged in both hospital

  19. Crystal structures of lysine-preferred racemases, the non-antibiotic selectable markers for transgenic plants.

    Directory of Open Access Journals (Sweden)

    Hsin-Mao Wu

    Full Text Available Lysine racemase, a pyridoxal 5'-phosphate (PLP-dependent amino acid racemase that catalyzes the interconversion of lysine enantiomers, is valuable to serve as a novel non-antibiotic selectable marker in the generation of transgenic plants. Here, we have determined the first crystal structure of a lysine racemase (Lyr from Proteus mirabilis BCRC10725, which shows the highest activity toward lysine and weaker activity towards arginine. In addition, we establish the first broad-specificity amino acid racemase (Bar structure from Pseudomonas putida DSM84, which presents not only the highest activity toward lysine but also remarkably broad substrate specificity. A complex structure of Bar-lysine is also established here. These structures demonstrate the similar fold of alanine racemase, which is a head-to-tail homodimer with each protomer containing an N-terminal (α/β(8 barrel and a C-terminal β-stranded domain. The active-site residues are located at the protomer interface that is a funnel-like cavity with two catalytic bases, one from each protomer, and the PLP binding site is at the bottom of this cavity. Structural comparisons, site-directed mutagenesis, kinetic, and modeling studies identify a conserved arginine and an adjacent conserved asparagine that fix the orientation of the PLP O3 atom in both structures and assist in the enzyme activity. Furthermore, side chains of two residues in α-helix 10 have been discovered to point toward the cavity and define the substrate specificity. Our results provide a structural foundation for the design of racemases with pre-determined substrate specificity and for the development of the non-antibiotic selection system in transgenic plants.

  20. Lysine requirement of growing male Pekin ducks.

    Science.gov (United States)

    Bons, A; Timmler, R; Jeroch, H

    2002-12-01

    1. One growth experiment and one balance test were conducted to study the response to increasing levels of dietary lysine supplementation in male Pekin ducks with special reference to the growth periods from 1 to 3 weeks and 4 to 7 weeks of age. 2. Two different low-lysine diets were used as basal diets in both periods. The basal lysine levels were 7.6 g/kg (d 1 to 21) and 6.2 g/kg (d 22 to 49) and the ranges in lysine concentration were 7.6 to 12.6 g/kg (d 1 to 21) and 6.2 to 11.2 g/kg (d 22 to 49). 3. Growth performance, feed conversion efficiency and meat yield increased (P < 0.05) with increasing lysine concentration (requirement defined as 95% of the asymptote). 4. It is concluded that the dietary lysine concentration should be 0.93 g/MJ nitrogen corrected apparent metabolisable energy (AMEN) (11.7 g/kg) for the starter period (until d 21) and 0.75 g/MJ AMEN (10.0 g/kg) for the grower period (from d 22 onwards).

  1. Structure of lysine adducts with 16 alpha-hydroxyestrone and cortisol.

    Science.gov (United States)

    Bucala, R; Ulrich, P C; Chait, B T; Bencsath, F A; Cerami, A

    1986-07-01

    Recent studies indicate that steroids containing a vicinal hydroxyketone moiety can react with proteins both in vitro and in vivo to form covalent addition products. This reaction is non-enzymatic and occurs via the Heyns rearrangement of an initial Schiff base adduct between the steroid carbonyl and the epsilon-amino group of lysine residues. The present study describes the synthesis, isolation, and structural analysis of model adducts prepared by the incubation of 16 alpha-hydroxyesterone or cortisol with NaCNBH3 and lysine derivatives blocked in the N alpha-position. The product formed from the reaction of 16 alpha-hydroxyesterone and lysine was found to have the structure predicted for a reduced Schiff base between these molecules. A stable, cortisol-lysine adduct was similarly synthesized and isolated. This conjugate was found not to be the expected reduced Schiff base but rather a C-20 cyano amine. This compound most likely was formed by the nucleophilic addition of cyanide during the course of the incubation. The observation that the cortisol-lysine Schiff base is not reducible with NaCNBH3 accounts for the observation that the incorporation rate of glucocorticoids into proteins is not increased by the presence of NaCNBH3.

  2. Differences in lysine pKa values may be used to improve NMR signal dispersion in reductively methylated proteins

    Energy Technology Data Exchange (ETDEWEB)

    Abraham, Sherwin J. [University of Illinois at Chicago, Department of Biochemistry and Molecular Genetics (United States); Kobayashi, Tomoyoshi; John Solaro, R. [University of Illinois at Chicago, Department of Physiology and Biophysics, Center for Cardiovascular Research (United States); Gaponenko, Vadim [University of Illinois at Chicago, Department of Biochemistry and Molecular Genetics (United States)], E-mail: vadimg@uic.edu

    2009-04-15

    Reductive methylation of lysine residues in proteins offers a way to introduce {sup 13}C methyl groups into otherwise unlabeled molecules. The {sup 13}C methyl groups on lysines possess favorable relaxation properties that allow highly sensitive NMR signal detection. One of the major limitations in the use of reductive methylation in NMR is the signal overlap of {sup 13}C methyl groups in NMR spectra. Here we show that the uniform influence of the solvent on chemical shifts of exposed lysine methyl groups could be overcome by adjusting the pH of the buffering solution closer to the pKa of lysine side chains. Under these conditions, due to variable pKa values of individual lysine side chains in the protein of interest different levels of lysine protonation are observed. These differences are reflected in the chemical shift differences of methyl groups in reductively methylated lysines. We show that this approach is successful in four different proteins including Ca{sup 2+}-bound Calmodulin, Lysozyme, Ca{sup 2+}-bound Troponin C, and Glutathione S-Transferase. In all cases significant improvement in NMR spectral resolution of methyl signals in reductively methylated proteins was obtained. The increased spectral resolution helps with more precise characterization of protein structural rearrangements caused by ligand binding as shown by studying binding of Calmodulin antagonist trifluoperazine to Calmodulin. Thus, this approach may be used to increase resolution in NMR spectra of {sup 13}C methyl groups on lysine residues in reductively methylated proteins that enhances the accuracy of protein structural assessment.

  3. Bioavailability of free lysine and protein-bound lysine from casein and fishmeal in juvenile turbot (Psetta maxima).

    Science.gov (United States)

    Kroeckel, Saskia; Dietz, Carsten; Schulz, Carsten; Susenbeth, Andreas

    2015-03-14

    In the present study, a linear regression analysis between lysine intake and lysine retention was conducted to investigate the efficiency of lysine utilisation (k(Lys)) at marginal lysine intake of either protein-bound or free lysine sources in juvenile turbot (Psetta maxima). For this purpose, nine isonitrogenous and isoenergetic diets were formulated to contain 2·25-4·12 g lysine/100 g crude protein (CP) to ensure that lysine was the first-limiting amino acid in all diets. The basal diet contained 2·25 g lysine/100 g CP. Graded levels of casein (Cas), fishmeal (FM) and L-lysine HCl (Lys) were added to the experimental diets to achieve stepwise lysine increments. A total of 240 fish (initial weight 50·1 g) were hand-fed all the experimental diets once daily until apparent satiation over a period of 56 d. Feed intake was significantly affected by dietary lysine concentration rather than by dietary lysine source. Specific growth rate increased significantly at higher lysine concentrations (PCas, Lys or FM were 0·833, 0·857 and 0·684, respectively. The bioavailability of lysine from the respective lysine sources was determined by a slope-ratio approach. The bioavailability of lysine (relative to the reference lysine source Cas) from FM and Lys was 82·1 and 103 %, respectively. Nutrient requirement for maintenance was in the range of 16·7-23·4 mg/kg(0·8) per d, and did not differ between the treatments. There were no significant differences in lysine utilisation efficiency or bioavailability of protein-bound or crystalline lysine from the respective sources observed when lysine was confirmed to be the first-limiting nutrient.

  4. Rapid analysis of multi-pesticide residues in lotus seeds by a modified QuEChERS-based extraction and GC-ECD.

    Science.gov (United States)

    Miao, Qing; Kong, Weijun; Yang, Shihai; Yang, Meihua

    2013-05-01

    A modified quick, easy, cheap, efficient, rugged and safe method (QuEChERS) coupled to gas chromatography with electron capture detector (GC-ECD) was developed for rapid extraction and simultaneous determination of 36 pesticides in lotus seeds. The extraction solvent (acetone, ethyl acetate, acetonitrile, n-hexane and n-hexane in combination with ethyl acetate) and purifying agent (neutral alumina, primary secondary amine, graphite carbon block and florisil) for QuEChERS extraction were optimized. The GC-ECD method was in-house validated in terms of linearity, selectivity, reproducibility, stability and recovery. The limits of detection (LODs) of the developed GC-ECD method for all investigated pesticides ranged from 0.01 to 3.0μgL(-1) and limits of quantification (LOQs) from 0.05 to 10.0μgL(-1). The satisfactory data demonstrated the good reproducibility and stability of the method with relative standard deviations (RSDs) lower than 15%. Recoveries for spiked lotus seed samples were from 60.84% to 119.91% with RSDs lower than 13.06%. Two out of 24 batches of lotus seeds collected in China were found to be contaminated with trans-chlordane, which were below LOQ. This is the first attempt in China using QuEChERS to GC-ECD to determine 36 major pesticides with differences in physio-chemical properties in lotus seeds. The method described here was found to be practicable in the routine residue analysis of pesticides in lotus seeds.

  5. Lysine-Rich Proteins in High-Lysine Hordeum Vulgare Grain

    DEFF Research Database (Denmark)

    Ingversen, J.; Køie, B.

    1973-01-01

    The salt-soluble proteins in barley grain selected for high-lysine content (Hiproly, CI 7115 and the mutants 29 and 86) and of a control (Carlsberg II) with normal lysine content, contain identical major proteins as determined by MW and electrophoretic mobility. The concentration of a protein group...

  6. A lysinated thiophene-based semiconductor as a multifunctional neural bioorganic interface.

    Science.gov (United States)

    Bonetti, Simone; Pistone, Assunta; Brucale, Marco; Karges, Saskia; Favaretto, Laura; Zambianchi, Massimo; Posati, Tamara; Sagnella, Anna; Caprini, Marco; Toffanin, Stefano; Zamboni, Roberto; Camaioni, Nadia; Muccini, Michele; Melucci, Manuela; Benfenati, Valentina

    2015-06-03

    Lysinated molecular organic semiconductors are introduced as valuable multifunctional platforms for neural cells growth and interfacing. Cast films of quaterthiophene (T4) semiconductor covalently modified with lysine-end moieties (T4Lys) are fabricated and their stability, morphology, optical/electrical, and biocompatibility properties are characterized. T4Lys films exhibit fluorescence and electronic transport as generally observed for unsubstituted oligothiophenes combined to humidity-activated ionic conduction promoted by the charged lysine-end moieties. The Lys insertion in T4 enables adhesion of primary culture of rat dorsal root ganglion (DRG), which is not achievable by plating cells on T4. Notably, on T4Lys, the number on adhering neurons/area is higher and displays a twofold longer neurite length than neurons plated on glass coated with poly-l-lysine. Finally, by whole-cell patch-clamp, it is shown that the biofunctionality of neurons cultured on T4Lys is preserved. The present study introduces an innovative concept for organic material neural interface that combines optical and iono-electronic functionalities with improved biocompatibility and neuron affinity promoted by Lys linkage and the softness of organic semiconductors. Lysinated organic semiconductors could set the scene for the fabrication of simplified bioorganic devices geometry for cells bidirectional communication or optoelectronic control of neural cells biofunctionality.

  7. Snorkeling of lysine side chains in transmembrane helices: how easy can it get?

    Science.gov (United States)

    Strandberg, Erik; Killian, J Antoinette

    2003-06-05

    Transmembrane segments of proteins are often flanked by lysine residues. The side chains of these residues may snorkel, i.e. they may bury themselves with their aliphatic part in the hydrophobic region of the lipid bilayer, while positioning the charged amino group in the more polar interface. Here we estimate the free energy cost of snorkeling from thermodynamical calculations based on studies with synthetic transmembrane peptides [Strandberg et al. (2002) Biochemistry 41, 7190-7198]. The value is estimated to be between 0.07 and 0.7 kcal mol(-1) for a lysine side chain. This very low value indicates that snorkeling may be a common process, which should be taken into consideration both in experimental and in theoretical studies on protein-lipid interactions.

  8. NMR determination of lysine pKa values in the Pol lambda lyase domain: mechanistic implications.

    Science.gov (United States)

    Gao, Guanghua; DeRose, Eugene F; Kirby, Thomas W; London, Robert E

    2006-02-14

    The base excision repair (BER) process requires removal of an abasic deoxyribose-5-phosphate group, a catalytic activity that has been demonstrated for the N-terminal 8 kDa domain of DNA polymerase beta (Pol beta), and for the homologous domain of DNA polymerase lambda (Pol lambda). Previous studies have demonstrated that this activity results from formation of a Schiff base adduct of the abasic deoxyribose C-1' with a lysine residue (K312 in the case of Pol lambda), followed by a beta-elimination reaction. To better understand the underlying chemistry, we have determined pKa values for the lysine residues in the Pol lambda lyase domain labeled with [epsilon-13C]lysine. At neutral pH, the H(epsilon) protons on 3 of the 10 lysine residues in this domain, K287, K291, and K312, exhibit chemical shift inequivalence that results from immobilization of the lysyl side chains. For K287 and K291, this results from the K287-E261 and K291-E298 salt bridge interactions, while for K312, immobilization apparently results from steric and hydrogen-bonding interactions that constrain the position of the lysine side chain. The pKa value of K312 is depressed to 9.58, a value indicating that at physiological pH K312 will exist predominantly in the protonated form. Titration of the domain with hairpin DNA containing a 5'-tetrahydrofuran terminus to model the abasic site produced shifts of the labeled lysine resonances that were in fast exchange but appeared to be complete at a stoichiometry of approximately 1:1.3, consistent with a dissociation constant of approximately 1 microM. The epsilon-proton shifts of K273 were the most sensitive to the addition of the DNA, apparently due to changes in the relative orientation between K273 and W274 in the DNA complex. The average pKa values increased by 0.55, consistent with the formation of some DNA-lysine salt bridges and with the general pH increase expected to result from a reduction in the net positive charge of the complex. A general

  9. Lysine methylation regulates the pRb tumour suppressor protein.

    Science.gov (United States)

    Munro, S; Khaire, N; Inche, A; Carr, S; La Thangue, N B

    2010-04-22

    The pRb tumour suppressor protein has a central role in coordinating early cell cycle progression. An important level of control imposed on pRb occurs through post-translational modification, for example, phosphorylation. We describe here a new level of regulation on pRb, mediated through the targeted methylation of lysine residues, by the methyltransferase Set7/9. Set7/9 methylates the C-terminal region of pRb, both in vitro and in cells, and methylated pRb interacts with heterochromatin protein HP1. pRb methylation is required for pRb-dependent cell cycle arrest and transcriptional repression, as well as pRb-dependent differentiation. Our results indicate that methylation can influence the properties of pRb, and raise the interesting possibility that methylation modulates pRb tumour suppressor activity.

  10. A mechanism-based potent sirtuin inhibitor containing Nε-thiocarbamoyl-lysine (TuAcK)

    OpenAIRE

    2011-01-01

    In the current study, we have identified Nε-thiocarbamoyl-lysine (TuAcK) as a general sirtuin inhibitory warhead which was shown to be able to confer potent sirtuin inhibition. This inhibition was also shown to be mechanism-based in that the TuAck residue was able to be processed by a sirtuin enzyme with the formation of a stalled S-alkylamidate intermediate.

  11. SET7/9 Catalytic Mutants Reveal the Role of Active Site Water Molecules in Lysine Multiple Methylation

    Energy Technology Data Exchange (ETDEWEB)

    Del Rizzo, Paul A.; Couture, Jean-François; Dirk, Lynnette M.A.; Strunk, Bethany S.; Roiko, Marijo S.; Brunzelle, Joseph S.; Houtz, Robert L.; Trievel, Raymond C. (Michigan); (NWU); (Kentucky)

    2010-11-15

    SET domain lysine methyltransferases (KMTs) methylate specific lysine residues in histone and non-histone substrates. These enzymes also display product specificity by catalyzing distinct degrees of methylation of the lysine {epsilon}-amino group. To elucidate the molecular mechanism underlying this specificity, we have characterized the Y245A and Y305F mutants of the human KMT SET7/9 (also known as KMT7) that alter its product specificity from a monomethyltransferase to a di- and a trimethyltransferase, respectively. Crystal structures of these mutants in complex with peptides bearing unmodified, mono-, di-, and trimethylated lysines illustrate the roles of active site water molecules in aligning the lysine {epsilon}-amino group for methyl transfer with S-adenosylmethionine. Displacement or dissociation of these solvent molecules enlarges the diameter of the active site, accommodating the increasing size of the methylated {epsilon}-amino group during successive methyl transfer reactions. Together, these results furnish new insights into the roles of active site water molecules in modulating lysine multiple methylation by SET domain KMTs and provide the first molecular snapshots of the mono-, di-, and trimethyl transfer reactions catalyzed by these enzymes.

  12. Deregulation of histone lysine methyltransferases contributes to oncogenic transformation of human bronchoepithelial cells

    Directory of Open Access Journals (Sweden)

    Yoda Satoshi

    2008-11-01

    Full Text Available Abstract Background Alterations in the processing of the genetic information in carcinogenesis result from stable genetic mutations or epigenetic modifications. It is becoming clear that nucleosomal histones are central to proper gene expression and that aberrant DNA methylation of genes and histone methylation plays important roles in tumor progression. To date, several histone lysine methyltransferases (HKMTs have been identified and histone lysine methylation is now considered to be a critical regulator of transcription. However, still relatively little is known about the role of HKMTs in tumorigenesis. Results We observed differential HKMT expression in a lung cancer model in which normal human bronchial epithelial (NHBE cells expressing telomerase, SV40 large T antigen, and Ras were immortal, formed colonies in soft agar, and expressed specific HKMTs for H3 lysine 9 and 27 residues but not for H3 lysine 4 residue. Modifications in the H3 tails affect the binding of proteins to the histone tails and regulate protein function and the position of lysine methylation marks a gene to be either activated or repressed. In the present study, suppression by siRNA of HKMTs (EZH2, G9A, SETDB1 and SUV39H1 that are over-expressed in immortalized and transformed cells lead to reduced cell proliferation and much less anchorage-independent colony growth. We also found that the suppression of H3-K9, G9A and SUV39H1 induced apoptosis and the suppression of H3-K27, EZH2 caused G1 arrest. Conclusion Our results indicate the potential of these HKMTs in addition to the other targets for epigenetics such as DNMTs and HDACs to be interesting therapeutic targets.

  13. A study on the effect of surface lysine to arginine mutagenesis on protein stability and structure using green fluorescent protein.

    Directory of Open Access Journals (Sweden)

    Sriram Sokalingam

    Full Text Available Two positively charged basic amino acids, arginine and lysine, are mostly exposed to protein surface, and play important roles in protein stability by forming electrostatic interactions. In particular, the guanidinium group of arginine allows interactions in three possible directions, which enables arginine to form a larger number of electrostatic interactions compared to lysine. The higher pKa of the basic residue in arginine may also generate more stable ionic interactions than lysine. This paper reports an investigation whether the advantageous properties of arginine over lysine can be utilized to enhance protein stability. A variant of green fluorescent protein (GFP was created by mutating the maximum possible number of lysine residues on the surface to arginines while retaining the activity. When the stability of the variant was examined under a range of denaturing conditions, the variant was relatively more stable compared to control GFP in the presence of chemical denaturants such as urea, alkaline pH and ionic detergents, but the thermal stability of the protein was not changed. The modeled structure of the variant indicated putative new salt bridges and hydrogen bond interactions that help improve the rigidity of the protein against different chemical denaturants. Structural analyses of the electrostatic interactions also confirmed that the geometric properties of the guanidinium group in arginine had such effects. On the other hand, the altered electrostatic interactions induced by the mutagenesis of surface lysines to arginines adversely affected protein folding, which decreased the productivity of the functional form of the variant. These results suggest that the surface lysine mutagenesis to arginines can be considered one of the parameters in protein stability engineering.

  14. Efficient Production of Enantiopure d-Lysine from l-Lysine by a Two-Enzyme Cascade System

    Directory of Open Access Journals (Sweden)

    Xin Wang

    2016-10-01

    Full Text Available The microbial production of d-lysine has been of great interest as a medicinal raw material. Here, a two-step process for d-lysine production from l-lysine by the successive microbial racemization and asymmetric degradation with lysine racemase and decarboxylase was developed. The whole-cell activities of engineered Escherichia coli expressing racemases from the strains Proteus mirabilis (LYR and Lactobacillus paracasei (AAR were first investigated comparatively. When the strain BL21-LYR with higher racemization activity was employed, l-lysine was rapidly racemized to give dl-lysine, and the d-lysine yield was approximately 48% after 0.5 h. Next, l-lysine was selectively catabolized to generate cadaverine by lysine decarboxylase. The comparative analysis of the decarboxylation activities of resting whole cells, permeabilized cells, and crude enzyme revealed that the crude enzyme was the best biocatalyst for enantiopure d-lysine production. The reaction temperature, pH, metal ion additive, and pyridoxal 5′-phosphate content of this two-step production process were subsequently optimized. Under optimal conditions, 750.7 mmol/L d-lysine was finally obtained from 1710 mmol/L l-lysine after 1 h of racemization reaction and 0.5 h of decarboxylation reaction. d-lysine yield could reach 48.8% with enantiomeric excess (ee ≥ 99%.

  15. Radioactive Lysine in Protein Metabolism Studies

    Science.gov (United States)

    Miller, L. L.; Bale, W. F.; Yuile, C. L.; Masters, R. E.; Tishkoff, G. H.; Whipple,, G. H.

    1950-01-09

    Studies of incorporation of DL-lysine in various body proteins of the dog; the time course of labeled blood proteins; and apparent rate of disappearance of labeled plasma proteins for comparison of behavior of the plasma albumin and globulin fractions; shows more rapid turn over of globulin fraction.

  16. Lysine and arginine requirements of Salminus brasiliensis

    Directory of Open Access Journals (Sweden)

    Jony Koji Dairiki

    2013-08-01

    Full Text Available The objective of this work was to determine the dietary lysine (DL and dietary arginine (DA requirements of dourado (Salminus brasiliensis, through dose-response trials using the amino acid profiles of whole carcasses as a reference. Two experiments were carried out in a completely randomized design (n=4. In the first experiment, groups of 12 feed-conditioned dourado juveniles (11.4±0.2 g were stocked in 60 L cages placed in 300 L plastic indoor tanks in a closed circulation system. Fish were fed for 60 days on diets containing 1.0, 1.5, 2.0, 2.5, 3.0, or 3.5 % dietary lysine. In the second experiment, dourado juveniles (27.0±0.8 g were fed for 60 days on semipurified diets containing arginine at 1.0, 1.5, 2.0, 2.5 or 3.0%, in similar conditions to those of the first experiment. Optimal DL requirements, as determined by broken-line analysis method for final weight, weight gain and specific growth rate, were 2.15% DL or 5% lysine in dietary protein, and 1.48% DA or 3.43% arginine in dietary protein. The best feed conversion ratio is attained with 2.5% DL or 5.8% lysine in dietary protein and 1.4% DA or 3.25% arginine in dietary protein.

  17. Desensitization to inhaled aztreonam lysine in an allergic patient with cystic fibrosis using a novel approach.

    Science.gov (United States)

    Guglani, Lokesh; Abdulhamid, Ibrahim; Ditouras, Joanna; Montejo, Jenny

    2012-10-01

    To report the successful desensitization of a highly allergic patient with cystic fibrosis (CF) to inhaled aztreonam lysine using the novel approach of intravenous desensitization followed by full-dose inhaled therapy without any adverse reactions. A 19-year-old woman with CF had persistent Pseudomonas aeruginosa-positive cultures and a history of type I hypersensitivity reactions to multiple medications, including aztreonam and tobramycin (intravenous and inhaled). To start therapy with an inhaled antipseudomonal antibiotic on a chronic basis, she underwent rapid desensitization to intravenous aztreonam followed by initiation of inhaled aztreonam lysine. Following intravenous desensitization with aztreonam, there was no adverse reaction or decline in lung function noted with inhaled aztreonam lysine and the chronic therapy was continued at home, with a modified regimen to maintain desensitization. Aztreonam lysine has been used for treatment of patients with CF with chronic P. aeruginosa colonization. Previous allergic reaction to intravenous aztreonam is considered a contraindication for use of aztreonam lysine. Our patient had a history of hives and facial swelling following administration of intravenous aztreonam (type I hypersensitivity reaction) as well as hypersensitivity to tobramycin. Rapid desensitization can be done for drugs that mediate a type I hypersensitivity reaction, with mast cells and basophils being the cellular targets. There are a few case reports of desensitization to inhaled antibiotics such as tobramycin and colistin, but desensitization to aztreonam lysine has not previously been reported. Desensitization of a patient with CF who is allergic to intravenous aztreonam was successfully accomplished with the novel approach of rapid intravenous desensitization followed by inhaled therapy. As inhaled antibiotics are being increasingly used for patients with CF, this novel strategy can be used for desensitizing allergic patients with CF to

  18. Lysine kinetics in preterm infants: the importance of enteral feeding

    NARCIS (Netherlands)

    S.R.D. van der Schoor (Sophie); P.J. Reeds; F. Stellaard; J.L.D. Wattimena (Josias); P.J.J. Sauer (Pieter); H.A. Büller (Hans); J.B. van Goudoever (Hans)

    2004-01-01

    textabstractINTRODUCTION: Lysine is the first limiting essential amino acid in the diet of newborns. First pass metabolism by the intestine of dietary lysine has a direct effect on systemic availability. We investigated whether first pass lysine metabolism in the intestine is high

  19. Lysine kinetics in preterm infants : the importance of enteral feeding

    NARCIS (Netherlands)

    van der Schoor, SRD; Reeds, PJ; Stellaard, F; Wattimena, JDL; Sauer, PJJ; Buller, HA; van Goudoever, JB

    2004-01-01

    Introduction: Lysine is the first limiting essential amino acid in the diet of newborns. First pass metabolism by the intestine of dietary lysine has a direct effect on systemic availability. We investigated whether first pass lysine metabolism in the intestine is high in preterm infants, particular

  20. The Presence of Modifiable Residues in the Core Peptide Part of Precursor Nisin Is Not Crucial for Precursor Nisin Interactions with NisB- and NisC

    NARCIS (Netherlands)

    Khusainov, Rustem; Kuipers, Oscar P.

    2013-01-01

    Precursor nisin is a model posttranslationally modified precursor lantibiotic that can be structurally divided into a leader peptide sequence and a modifiable core peptide part. The nisin core peptide clearly plays an important role in the precursor nisin - nisin modification enzymes interactions, s

  1. Saturation mutagenesis of lysine 12 leads to the identification of derivatives of nisin A with enhanced antimicrobial activity.

    Directory of Open Access Journals (Sweden)

    Evelyn M Molloy

    Full Text Available It is becoming increasingly apparent that innovations from the "golden age" of antibiotics are becoming ineffective, resulting in a pressing need for novel therapeutics. The bacteriocin family of antimicrobial peptides has attracted much attention in recent years as a source of potential alternatives. The most intensively studied bacteriocin is nisin, a broad spectrum lantibiotic that inhibits gram-positive bacteria including important food pathogens and clinically relevant antibiotic resistant bacteria. Nisin is gene-encoded and, as such, is amenable to peptide bioengineering, facilitating the generation of novel derivatives that can be screened for desirable properties. It was to this end that we used a site-saturation mutagenesis approach to create a bank of producers of nisin A derivatives that differ with respect to the identity of residue 12 (normally lysine; K12. A number of these producers exhibited enhanced bioactivity and the nisin A K12A producer was deemed of greatest interest. Subsequent investigations with the purified antimicrobial highlighted the enhanced specific activity of this modified nisin against representative target strains from the genera Streptococcus, Bacillus, Lactococcus, Enterococcus and Staphylococcus.

  2. Production of L-lysine on different silage juices using genetically engineered Corynebacterium glutamicum.

    Science.gov (United States)

    Neuner, Andreas; Wagner, Ines; Sieker, Tim; Ulber, Roland; Schneider, Konstantin; Peifer, Susanne; Heinzle, Elmar

    2013-01-20

    Corynebacterium glutamicum, the best established industrial producer organism for lysine was genetically modified to allow the production of lysine on grass and corn silages. The resulting strain C. glutamicum lysC(fbr)dld(Psod)pyc(Psod)malE(Psod)fbp(Psod)gapX(Psod) was based on earlier work (Neuner and Heinzle, 2011). That mutant carries a point mutation in the aspartokinase (lysC) regulatory subunit gene as well as overexpression of D-lactate dehydrogenase (dld), pyruvate carboxylase (pyc) and malic enzyme (malE) using the strong Psod promoter. Here, we additionally overexpressed fructose 1,6-bisphosphatase (fbp) and glyceraldehyde 3-phosphate dehydrogenase (gapX) using the same promoter. The resulting strain grew readily on grass and corn silages with a specific growth rate of 0.35 h⁻¹ and lysine carbon yields of approximately 90 C-mmol (C-mol)⁻¹. Lysine yields were hardly affected by oxygen limitation whereas linear growth was observed under oxygen limiting conditions. Overall, this strain seems very robust with respect to the composition of silage utilizing all quantified low molecular weight substrates, e.g. lactate, glucose, fructose, maltose, quinate, fumarate, glutamate, leucine, isoleucine and alanine.

  3. Interaction of L-lysine and soluble elastin with the semicarbazide-sensitive amine oxidase in the context of its vascular-adhesion and tissue maturation functions.

    LENUS (Irish Health Repository)

    Olivieri, Aldo

    2010-04-01

    The copper-containing quinoenzyme semicarbazide-sensitive amine oxidase (EC 1.4.3.21; SSAO) is a multifunctional protein. In some tissues, such as the endothelium, it also acts as vascular-adhesion protein 1 (VAP-1), which is involved in inflammatory responses and in the chemotaxis of leukocytes. Earlier work had suggested that lysine might function as a recognition molecule for SSAO\\/VAP-1. The present work reports the kinetics of the interaction of L-lysine and some of its derivatives with SSAO. Binding was shown to be saturable, time-dependent but reversible and to cause uncompetitive inhibition with respect to the amine substrate. It was also specific, since D-lysine, L-lysine ethyl ester and epsilon-acetyl-L-lysine, for example, did not bind to the enzyme. The lysine-rich protein soluble elastin bound to the enzyme relatively tightly, which may have relevance to the reported roles of SSAO in maintaining the extracellular matrix (ECM) and in the maturation of elastin. Our data show that lysyl residues are not oxidized by SSAO, but they bind tightly to the enzyme in the presence of hydrogen peroxide. This suggests that binding in vivo of SSAO to lysyl residues in physiological targets might be regulated in the presence of H(2)O(2), formed during the oxidation of a physiological SSAO substrate, yet to be identified.

  4. 石棉尾矿酸浸渣填充改性道路沥青的研究%Research on Road Asphalt Filled and Modified with Acid-leaching Residue of Asbestos Tailings

    Institute of Scientific and Technical Information of China (English)

    孙志明; 郑水林; 文明; 吴照洋

    2009-01-01

    The acid-leaching residue of asbestos tailings is silicon slag of asbestos tailings, one kind of solid waste after acid leaching extraction of magnesium. In this experiment, using acid leaching residue of asbestos tailings after calcination as the filler of asphalt modifier, through the test of penetration, ductility, softening point of modified asphalt, the effects of the adding volume and mixing conditions (temperature, time, etc.) of the acid leaching residue on the performance of road asphalt has been studied. The results showed that the appropriate conditions of modified process is that the adding volume of acid leaching residue 6%, mixing temperature 140℃, and heating mixing time 20rain. In this condition, the performance of modified asphalt material, such as high temperature and low temperature performance, temperature sensitivity, and anti-aging property have been significantly improved.%石棉尾矿酸浸渣是石棉尾矿蛇纹石酸浸提取氧化镁后的硅质粉体材料.本实验以煅烧后的石棉尾矿酸浸渣作为道路沥青填充改性剂,通过测定改性沥青的针入度、延度、软化点等指标,研究了酸浸渣填充量、混合控温以及加热混合时间对道路沥青综合性能的影响.结果表明,石棉尾矿酸浸渣填充改性沥青适宜的填充工艺条件为填充量6%,混合控温140℃,加热混合时间20min;在适宜的填充工艺条件下,改性沥青的高温性能、低温性能、高低温稳定性以及抗老化性均得到显著改善或提高.

  5. Expression and purification of histone H3 proteins containing multiple sites of lysine acetylation using nonsense suppression.

    Science.gov (United States)

    Young, Isaac A; Mittal, Chitvan; Shogren-Knaak, Michael A

    2016-02-01

    Lysine acetylation is a common post-translational modification, which is especially prevalent in histone proteins in chromatin. A number of strategies exist for generating histone proteins containing lysine acetylation, but an especially attractive approach is to genetically encode acetyl-lysine residues using nonsense suppression. This strategy has been successfully applied to single sites of histone acetylation. However, because histone acetylation can often occur at multiple sites simultaneously, we were interested in determining whether this approach could be extended. Here we show that we can express histone H3 proteins that incorporate up to four sites of lysine acetylation on the histone tail. Because the amount of expressed multi-acetylated histone is reduced relative to the wild type, a purification strategy involving affinity purification and ion exchange chromatography was optimized. This expression and purification strategy ultimately generates H3 histone uniformly acetylated at the desired position at levels and purity sufficient to assemble histone octamers. Histone octamers containing four sites of lysine acetylation were assembled into mononucleosomes and enzymatic assays confirmed that this acetylation largely blocks further acetylation by the yeast SAGA acetyltransferase complex.

  6. Lysine conservation and context in TGFbeta and Wnt signaling suggest new targets and general themes for posttranslational modification.

    Science.gov (United States)

    Konikoff, Charlotte E; Wisotzkey, Robert G; Newfeld, Stuart J

    2008-10-01

    TGFbeta and Wnt pathways play important roles in the development of animals from sponges to humans. In both pathways posttranslational modification as a means of regulating their function, such as lysine modification by ubiquitination and sumoylation, has been observed. However, a gap exists between the immunological observation of posttranslational modification and the identification of the target lysine. To fill this gap, we conducted a phylogenetic analysis of lysine conservation and context in TGFbeta and Wnt pathway receptors and signal transducers and suggest numerous high-probability candidates for posttranslational modification. Further comparison of results from both pathways suggests two general features for biochemical regulation of intercellular signaling: receptors are less frequent targets for modification than signal transduction agonists, and a lysine adjacent to an upstream hydrophobic residue may be a preferred context for modification. Overall the results suggest numerous applications for an evolutionary approach to the biochemical regulation of developmental pathways, including (1) streamlining of the identification of the target lysine, (2) determination of when members of a multigene family acquire distinct activities, (3) application to any conserved protein family, and (4) application to any modification of a specific amino acid.

  7. Differences in lysine adduction by acrolein and methyl vinyl ketone: implications for cytotoxicity in cultured hepatocytes.

    Science.gov (United States)

    Kaminskas, Lisa M; Pyke, Simon M; Burcham, Philip C

    2005-11-01

    Acrolein is a highly toxic environmental pollutant that readily alkylates the epsilon-amino group of lysine residues in proteins. In model systems, such chemistry involves sequential addition of two acrolein molecules to a given nitrogen, forming bis-Michael-adducted species that undergo aldol condensation and dehydration to form Nepsilon-(3-formyl-3,4-dehydropiperidino)lysine. Whether this ability to form cyclic adducts participates in the toxicity of acrolein is unknown. To address this issue, we compared the chemistry of protein adduction by acrolein to that of its close structural analogue methyl vinyl ketone, expecting that the alpha-methyl group would hinder the intramolecular cyclization of any bis-adducted species formed by methyl vinyl ketone. Both acrolein and methyl vinyl ketone displayed comparable protein carbonylating activity during in vitro studies with the model protein bovine serum albumin, confirming the alpha,beta,-unsaturated bond of both compounds is an efficient Michael acceptor for protein nucleophiles. However, differences in adduction chemistry became apparent during the use of electrospray ionization-MS to monitor reaction products in a lysine-containing peptide after modification by each compound. For example, although a Schiff base adduct was detected following reaction of the peptide with acrolein, an analogous species was not formed by methyl vinyl ketone. Furthermore, while ions corresponding to mono- and bis-Michael adducts were detected at the N-terminus and lysine residues following peptide modification by both carbonyls, only acrolein modification generated ions attributable to cyclic adducts. Despite these differences in adduction chemistry, in mouse hepatocytes, the two compounds exhibited very comparable abilities to induce rapid, concentration-dependent cell death as well as protein carbonylation. These findings suggest that the acute toxicity of short-chain alpha,beta-unsaturated carbonyl compounds involves their ability to

  8. Catalytic roles of lysines (K9, K27, K31) in the N-terminal domain in human adenylate kinase by random site-directed mutagenesis.

    Science.gov (United States)

    Ayabe, T; Park, S K; Takenaka, H; Sumida, M; Uesugi, S; Takenaka, O; Hamada, M

    1996-11-01

    To elucidate lysine residues in the N-terminal domain of human cytosolic adenylate kinase (hAK1, EC 2.7.4.3), random site-directed mutagenesis of K9, K27, and K31 residues was performed, and six mutants were analyzed by steady-state kinetics. K9 residue may play an important role in catalysis by interacting with AMP2-. K27 and K31 residues appear to play a functional role in catalysis by interacting with MgATP2-. In human AK, the epsilon-amino group in the side chain of these lysine residues would be essential for phosphoryl transfer between MgATP2- and AMP2- during transition state.

  9. Lysine fortification: past, present, and future.

    Science.gov (United States)

    Pellett, Peter L; Ghosh, Shibani

    2004-06-01

    Fortification with lysine to improve the protein value of human diets that are heavily based on cereals has received support from the results of these recent studies [1,2]. Support also comes from examination of average food and nutrient availability data derived from food balance sheets. Whereas nutritional status is influenced by the nutrient content of foods consumed in relation to need, the requirements for protein and amino acids are influenced by many additional factors [10, 12, 14, 28, 29]. These include age, sex, body size, physical activity, growth, pregnancy and lactation, infection, and the efficiency of nutrient utilization. Even if the immune response was influenced by the added lysine, adequate water and basic sanitation would remain essential. Acute and chronic undernutrition and most micronutrient deficiencies primarily affect poor and deprived people who do not have access to food of adequate nutritional value, live in unsanitary environments without access to clean water and basic services, and lack access to appropriate education and information [30]. A further variable is the possible interaction between protein and food energy availability [31]. This could affect the protein value of diets when food energy is limiting to a significant degree. Thus, the additional effects of food energy deficiency on protein utilization could well be superimposed on the very poorest. The improvement of dietary diversity must be the long-term aim, with dietary fortification considered only a short-term solution. The former should take place as wealth improves and the gaps between rich and poor diminish. Although such changes are taking place, they are highly uneven. Over the last several decades, increases have occurred in the availability of food energy, total protein, and animal protein for both developed and developing countries. However, for the very poorest developing countries over the same period, changes have been almost nonexistent, and the values for

  10. Charge neutralization of the central lysine cluster in prion protein (PrP) promotes PrP(Sc)-like folding of recombinant PrP amyloids.

    Science.gov (United States)

    Groveman, Bradley R; Kraus, Allison; Raymond, Lynne D; Dolan, Michael A; Anson, Kelsie J; Dorward, David W; Caughey, Byron

    2015-01-09

    The structure of the infectious form of prion protein, PrP(Sc), remains unclear. Most pure recombinant prion protein (PrP) amyloids generated in vitro are not infectious and lack the extent of the protease-resistant core and solvent exclusion of infectious PrP(Sc), especially within residues ∼90-160. Polyanionic cofactors can enhance infectivity and PrP(Sc)-like characteristics of such fibrils, but the mechanism of this enhancement is unknown. In considering structural models of PrP(Sc) multimers, we identified an obstacle to tight packing that might be overcome with polyanionic cofactors, namely, electrostatic repulsion between four closely spaced cationic lysines within a central lysine cluster of residues 101-110. For example, in our parallel in-register intermolecular β-sheet model of PrP(Sc), not only would these lysines be clustered within the 101-110 region of the primary sequence, but they would have intermolecular spacings of only ∼4.8 Å between stacked β-strands. We have now performed molecular dynamics simulations predicting that neutralization of the charges on these lysine residues would allow more stable parallel in-register packing in this region. We also show empirically that substitution of these clustered lysine residues with alanines or asparagines results in recombinant PrP amyloid fibrils with extended proteinase-K resistant β-sheet cores and infrared spectra that are more reminiscent of bona fide PrP(Sc). These findings indicate that charge neutralization at the central lysine cluster is critical for the folding and tight packing of N-proximal residues within PrP amyloid fibrils. This charge neutralization may be a key aspect of the mechanism by which anionic cofactors promote PrP(Sc) formation.

  11. Influence of shifting positions of Ser, Thr, and Cys residues in prenisin on the efficiency of modification reactions and on the antimicrobial activities of the modified prepeptides

    NARCIS (Netherlands)

    Lubelski, Jacek; Overkamp, Wout; Kluskens, Leon D.; Moll, Gert N.; Kuipers, Oscar P.

    2008-01-01

    Since the recent discovery that the nisin modification and transport machinery can be used to produce and modify peptides unrelated to nisin, specific questions arose concerning the specificity of the modification enzymes involved and the limits of their promiscuity with respect to the dehydration a

  12. Exploring lysine riboswitch for metabolic flux control and improvement of L-lysine synthesis in Corynebacterium glutamicum.

    Science.gov (United States)

    Zhou, Li-Bang; Zeng, An-Ping

    2015-06-19

    Riboswitch, a regulatory part of an mRNA molecule that can specifically bind a metabolite and regulate gene expression, is attractive for engineering biological systems, especially for the control of metabolic fluxes in industrial microorganisms. Here, we demonstrate the use of lysine riboswitch and intracellular l-lysine as a signal to control the competing but essential metabolic by-pathways of lysine biosynthesis. To this end, we first examined the natural lysine riboswitches of Eschericia coli (ECRS) and Bacillus subtilis (BSRS) to control the expression of citrate synthase (gltA) and thus the metabolic flux in the tricarboxylic acid (TCA) cycle in E. coli. ECRS and BSRS were then successfully used to control the gltA gene and TCA cycle activity in a lysine producing strain Corynebacterium glutamicum LP917, respectively. Compared with the strain LP917, the growth of both lysine riboswitch-gltA mutants was slower, suggesting a reduced TCA cycle activity. The lysine production was 63% higher in the mutant ECRS-gltA and 38% higher in the mutant BSRS-gltA, indicating a higher metabolic flux into the lysine synthesis pathway. This is the first report on using an amino acid riboswitch for improvement of lysine biosynthesis. The lysine riboswitches can be easily adapted to dynamically control other essential but competing metabolic pathways or even be engineered as an "on-switch" to enhance the metabolic fluxes of desired metabolic pathways.

  13. Antimicrobial activity of chicken NK-lysin against Eimeria sporozoites.

    Science.gov (United States)

    Hong, Yeong H; Lillehoj, Hyun S; Siragusa, Gregory R; Bannerman, Douglas D; Lillehoj, Erik P

    2008-06-01

    NK-lysin is an antimicrobial and antitumor polypeptide that is considered to play an important role in innate immunity. Chicken NK-lysin is a member of the saposin-like protein family and exhibits potent antitumor cell activity. To evaluate the antimicrobial properties of chicken NK-lysin, we examined its ability to reduce the viability of various bacterial strains and two species of Eimeria parasites. Culture supernatants from COS7 cells transfected with a chicken NK-lysin cDNA and His-tagged purified NK-lysin from the transfected cells both showed high cytotoxic activity against Eimeria acervulina and Eimeria maxima sporozoites. In contrast, no bactericidal activity was observed. Further studies using synthetic peptides derived from NK-lysin may be useful for pharmaceutical and agricultural uses in the food animal industry.

  14. Global analysis of lysine acetylation in strawberry leaves

    Directory of Open Access Journals (Sweden)

    Xianping eFang

    2015-09-01

    Full Text Available Protein lysine acetylation is a reversible and dynamic post-translational modification. It plays an important role in regulating diverse cellular processes including chromatin dynamic, metabolic pathways and transcription in both prokaryotes and eukaryotes. Although studies of lysine acetylome in plants have been reported, the throughput was not high enough, hindering the deep understanding of lysine acetylation in plant physiology and pathology. In this study, taking advantages of anti-acetyllysine-based enrichment and high-sensitive-mass spectrometer, we applied an integrated proteomic approach to comprehensively investigate lysine acetylome in strawberry. In total, we identified 1392 acetylation sites in 684 proteins, representing the largest dataset of acetylome in plants to date. To reveal the functional impacts of lysine acetylation in strawberry, intensive bioinformatic analysis was performed. The results significantly expanded our current understanding of plant acetylome and demonstrated that lysine acetylation is involved in multiple cellular metabolism and cellular processes. More interestingly, nearly 50% of all acetylated proteins identified in this work were localized in chloroplast and the vital role of lysine acetylation in photosynthesis was also revealed. Taken together, this study not only established the most extensive lysine acetylome in plants to date, but also systematically suggests the significant and unique roles of lysine acetylation in plants.

  15. Acrolein modification impairs key functional features of rat apolipoprotein E: identification of modified sites by mass spectrometry.

    Science.gov (United States)

    Tran, Tuyen N; Kosaraju, Malathi G; Tamamizu-Kato, Shiori; Akintunde, Olayemi; Zheng, Ying; Bielicki, John K; Pinkerton, Kent; Uchida, Koji; Lee, Yuan Yu; Narayanaswami, Vasanthy

    2014-01-21

    Apolipoprotein E (apoE), an antiatherogenic apolipoprotein, plays a significant role in the metabolism of lipoproteins. It lowers plasma lipid levels by acting as a ligand for the low-density lipoprotein receptor (LDLr) family of proteins, in addition to playing a role in promoting macrophage cholesterol efflux in atherosclerotic lesions. The objective of this study is to examine the effect of acrolein modification on the structure and function of rat apoE and to determine the sites and nature of modification by mass spectrometry. Acrolein is a highly reactive aldehyde, which is generated endogenously as one of the products of lipid peroxidation and is present in the environment in pollutants such as tobacco smoke and heated oils. In initial studies, acrolein-modified apoE was identified by immunoprecipitation using an acrolein-lysine specific antibody in the plasma of 10-week old male rats that were exposed to filtered air (FA) or low doses of environmental tobacco smoke (ETS). While both groups displayed acrolein-modified apoE in the lipoprotein fraction, the ETS group had higher levels in the lipid-free fraction compared with the FA group. This observation provided the rationale to further investigate the effect of acrolein modification on rat apoE at a molecular level. Treatment of recombinant rat apoE with a 10-fold molar excess of acrolein resulted in (i) a significant decrease in lipid-binding and cholesterol efflux abilities, (ii) impairment in the LDLr- and heparin-binding capabilities, and (iii) significant alterations in the overall stability of the protein. The disruption in the functional abilities is attributed directly or indirectly to acrolein modification yielding an aldimine adduct at K149 and K155 (+38); a propanal adduct at K135 and K138 (+56); an N(ε)-(3-methylpyridinium)lysine (MP-lysine) at K64, K67, and K254 (+76), and an N(ε)-(3-formyl-3,4-dehydropiperidino)lysine (FDP-lysine) derivative at position K68 (+94), as determined by matrix

  16. Histone H4 Lysine 20 methylation

    DEFF Research Database (Denmark)

    Jørgensen, Stine; Schotta, Gunnar; Sørensen, Claus Storgaard

    2013-01-01

    of histones have emerged as key regulators of genomic integrity. Intense research during the past few years has revealed histone H4 lysine 20 methylation (H4K20me) as critically important for the biological processes that ensure genome integrity, such as DNA damage repair, DNA replication and chromatin...... instability, demonstrating the important functions of H4K20 methylation in genome maintenance. In this review, we explain molecular mechanisms underlying these defects and discuss novel ideas for furthering our understanding of genome maintenance in higher eukaryotes....

  17. Optimization of lysine metabolism in Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Rytter, Jakob Vang

    the project intends to eliminate. PGI catalyzes the conversion of alpha-D-glucose-6-phosphate to fructose-6-phosphate just downstream of the branch in the glycolysis, but it also catalyzes the reverse reaction. It is unknown whether up- or down-regulation of the pgi is required to increase the flux through......, and increased NADPH availability is therefore a potential way to enhance lysine production. The generation of NADPH is mainly located in the pentose phosphate pathway (PPP). Using the genome scale model the phosphoglucoisomerase enzyme (PGI) has been identified as a possible bottleneck in the metabolism, which...

  18. Chromosomal protein HMGN1 enhances the acetylation of lysine 14 in histone H3

    OpenAIRE

    Lim, Jae-Hwan; West, Katherine L.; Rubinstein, Yaffa; Bergel, Michael; Postnikov, Yuri V.; Bustin, Michael

    2005-01-01

    The acetylation levels of lysine residues in nucleosomes, which are determined by the opposing activities of histone acetyltransferases (HATs) and deacetylases, play an important role in regulating chromatin-related processes, including transcription. We report that HMGN1, a nucleosomal binding protein that reduces the compaction of the chromatin fiber, increases the levels of acetylation of K14 in H3. The levels of H3K14ac in Hmgn1−/− cells are lower than in Hmgn1+/+ cells. Induced expressio...

  19. Triple therapy with pyridoxine, arginine supplementation and dietary lysine restriction in pyridoxine-dependent epilepsy: Neurodevelopmental outcome.

    Science.gov (United States)

    Coughlin, Curtis R; van Karnebeek, Clara D M; Al-Hertani, Walla; Shuen, Andrew Y; Jaggumantri, Sravan; Jack, Rhona M; Gaughan, Sommer; Burns, Casey; Mirsky, David M; Gallagher, Renata C; Van Hove, Johan L K

    2015-01-01

    Pyridoxine-dependent epilepsy (PDE) is an epileptic encephalopathy characterized by response to pharmacologic doses of pyridoxine. PDE is caused by deficiency of α-aminoadipic semialdehyde dehydrogenase resulting in impaired lysine degradation and subsequent accumulation of α-aminoadipic semialdehyde. Despite adequate seizure control with pyridoxine monotherapy, 75% of individuals with PDE have significant developmental delay and intellectual disability. We describe a new combined therapeutic approach to reduce putative toxic metabolites from impaired lysine metabolism. This approach utilizes pyridoxine, a lysine-restricted diet to limit the substrate that leads to neurotoxic metabolite accumulation and L-arginine to compete for brain lysine influx and liver mitochondrial import. We report the developmental and biochemical outcome of six subjects who were treated with this triple therapy. Triple therapy reduced CSF, plasma, and urine biomarkers associated with neurotoxicity in PDE. The addition of arginine supplementation to children already treated with dietary lysine restriction and pyridoxine further reduced toxic metabolites, and in some subjects appeared to improve neurodevelopmental outcome. Dietary lysine restriction was associated with improved seizure control in one subject, and the addition of arginine supplementation increased the objective motor outcome scale in two twin siblings, illustrating the contribution of each component of this treatment combination. Optimal results were noted in the individual treated with triple therapy early in the course of the disease. Residual disease symptoms could be related to early injury suggested by initial MR imaging prior to initiation of treatment or from severe epilepsy prior to diagnosis. This observational study reports the use of triple therapy, which combines three effective components in this rare condition, and suggests that early diagnosis and treatment with this new triple therapy may ameliorate the

  20. File list: Oth.Unc.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Unc.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Unclassified ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Unc.50.Crotonyl_lysine.AllCell.bed ...

  1. File list: Oth.Pan.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Pan.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Pancreas http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Pan.05.Crotonyl_lysine.AllCell.bed ...

  2. File list: Oth.Plc.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Plc.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Placenta http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Plc.20.Crotonyl_lysine.AllCell.bed ...

  3. File list: Oth.Unc.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Unc.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Unclassified ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Unc.10.Crotonyl_lysine.AllCell.bed ...

  4. File list: Oth.Unc.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Unc.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Unclassified ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Unc.20.Crotonyl_lysine.AllCell.bed ...

  5. File list: Oth.Pan.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Pan.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Pancreas http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Pan.50.Crotonyl_lysine.AllCell.bed ...

  6. File list: Oth.Plc.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Plc.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Placenta http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Plc.50.Crotonyl_lysine.AllCell.bed ...

  7. File list: Oth.Prs.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Prs.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Prostate http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Prs.10.Crotonyl_lysine.AllCell.bed ...

  8. File list: Oth.Prs.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Prs.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Prostate http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Prs.05.Crotonyl_lysine.AllCell.bed ...

  9. File list: Oth.Prs.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Prs.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Prostate http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Prs.20.Crotonyl_lysine.AllCell.bed ...

  10. File list: Oth.Plc.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Plc.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Placenta http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Plc.05.Crotonyl_lysine.AllCell.bed ...

  11. File list: Oth.Pan.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Pan.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Pancreas http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Pan.10.Crotonyl_lysine.AllCell.bed ...

  12. File list: Oth.Plc.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Plc.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Placenta http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Plc.10.Crotonyl_lysine.AllCell.bed ...

  13. File list: Oth.Prs.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Prs.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Prostate http...://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Prs.50.Crotonyl_lysine.AllCell.bed ...

  14. File list: Oth.Unc.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Unc.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Unclassified ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Unc.05.Crotonyl_lysine.AllCell.bed ...

  15. Quantification of Lysine Acetylation and Succinylation Stoichiometry in Proteins Using Mass Spectrometric Data-Independent Acquisitions (SWATH).

    Science.gov (United States)

    Meyer, Jesse G; D'Souza, Alexandria K; Sorensen, Dylan J; Rardin, Matthew J; Wolfe, Alan J; Gibson, Bradford W; Schilling, Birgit

    2016-11-01

    Post-translational modification of lysine residues by NƐ-acylation is an important regulator of protein function. Many large-scale protein acylation studies have assessed relative changes of lysine acylation sites after antibody enrichment using mass spectrometry-based proteomics. Although relative acylation fold-changes are important, this does not reveal site occupancy, or stoichiometry, of individual modification sites, which is critical to understand functional consequences. Recently, methods for determining lysine acetylation stoichiometry have been proposed based on ratiometric analysis of endogenous levels to those introduced after quantitative per-acetylation of proteins using stable isotope-labeled acetic anhydride. However, in our hands, we find that these methods can overestimate acetylation stoichiometries because of signal interferences when endogenous levels of acylation are very low, which is especially problematic when using MS1 scans for quantification. In this study, we sought to improve the accuracy of determining acylation stoichiometry using data-independent acquisition (DIA). Specifically, we use SWATH acquisition to comprehensively collect both precursor and fragment ion intensity data. The use of fragment ions for stoichiometry quantification not only reduces interferences but also allows for determination of site-level stoichiometry from peptides with multiple lysine residues. We also demonstrate the novel extension of this method to measurements of succinylation stoichiometry using deuterium-labeled succinic anhydride. Proof of principle SWATH acquisition studies were first performed using bovine serum albumin for both acetylation and succinylation occupancy measurements, followed by the analysis of more complex samples of E. coli cell lysates. Although overall site occupancy was low (<1%), some proteins contained lysines with relatively high acetylation occupancy. Graphical Abstract ᅟ.

  16. Quantification of Lysine Acetylation and Succinylation Stoichiometry in Proteins Using Mass Spectrometric Data-Independent Acquisitions (SWATH)

    Science.gov (United States)

    Meyer, Jesse G.; D'Souza, Alexandria K.; Sorensen, Dylan J.; Rardin, Matthew J.; Wolfe, Alan J.; Gibson, Bradford W.; Schilling, Birgit

    2016-09-01

    Post-translational modification of lysine residues by NƐ-acylation is an important regulator of protein function. Many large-scale protein acylation studies have assessed relative changes of lysine acylation sites after antibody enrichment using mass spectrometry-based proteomics. Although relative acylation fold-changes are important, this does not reveal site occupancy, or stoichiometry, of individual modification sites, which is critical to understand functional consequences. Recently, methods for determining lysine acetylation stoichiometry have been proposed based on ratiometric analysis of endogenous levels to those introduced after quantitative per-acetylation of proteins using stable isotope-labeled acetic anhydride. However, in our hands, we find that these methods can overestimate acetylation stoichiometries because of signal interferences when endogenous levels of acylation are very low, which is especially problematic when using MS1 scans for quantification. In this study, we sought to improve the accuracy of determining acylation stoichiometry using data-independent acquisition (DIA). Specifically, we use SWATH acquisition to comprehensively collect both precursor and fragment ion intensity data. The use of fragment ions for stoichiometry quantification not only reduces interferences but also allows for determination of site-level stoichiometry from peptides with multiple lysine residues. We also demonstrate the novel extension of this method to measurements of succinylation stoichiometry using deuterium-labeled succinic anhydride. Proof of principle SWATH acquisition studies were first performed using bovine serum albumin for both acetylation and succinylation occupancy measurements, followed by the analysis of more complex samples of E. coli cell lysates. Although overall site occupancy was low (<1%), some proteins contained lysines with relatively high acetylation occupancy.

  17. Determination of Residual Nonsteroidal Anti-Inflammatory Drugs in Aqueous Sample Using Magnetic Nanoparticles Modified with Cetyltrimethylammonium Bromide by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Malihe Khoeini Sharifabadi

    2014-01-01

    Full Text Available A simple and sensitive solid-phase extraction method for separation and preconcentration of trace amount of four nonsteroidal anti-inflammatory drugs (naproxen, indomethacin, diclofenac, and ibuprofen using Fe3O4 magnetic nanoparticles modified with cetyltrimethylammonium bromide has been developed. For this purpose, the surface of MNPs was modified with cetyltrimethylammonium bromide (CTAB as a cationic surfactant. Effects of different parameters influencing the extraction efficiency of drugs including the pH, amount of salt, shaking time, eluent type, the volume of solvent, amount of adsorbent, sample volume, and the time of desorption were investigated and optimized. Methanol has been used as desorption solvent and the extracts were analysed on a reversed-phase octadecyl silica column using 0.02 M phosphate-buffer (pH = 6.02 acetonitrile (65 : 35 v/v as the mobile phase and the effluents were measured at 202 nm with ultraviolet detector. The relative standard deviation (RSD% of the method was investigated at three concentrations (25, 50, and 200 ng/mL and was in the range of 3.98–9.83% (n=6 for 50 ng/mL. The calibration curves obtained for studied drugs show reasonable linearity (R2>0.99 and the limit of detection (LODs ranged between 2 and 7 ng/mL. Finally, the proposed method has been effectively employed in extraction and determination of the drugs in biological and environmental samples.

  18. Quantification of Nε-(2-Furoylmethyl)-L-lysine (furosine), Nε-(Carboxymethyl)-L-lysine (CML), Nε-(Carboxyethyl)-L-lysine (CEL) and total lysine through stable isotope dilution assay and tandem mass spectrometry

    NARCIS (Netherlands)

    Troise, A.D.; Fiore, A.; Wiltafsky, M.; Fogliano, V.

    2015-01-01

    The control of Maillard reaction (MR) is a key point to ensure processed foods quality. Due to the presence of a primary amino group on its side chain, lysine is particularly prone to chemical modifications with the formation of Amadori products (AP), Nε-(Carboxymethyl)-L-lysine (CML),

  19. STUDY OF LYSINE AND ALANINE DELIVERANCE THROUGH POLYPYRROLE MEMBRANE

    Directory of Open Access Journals (Sweden)

    Adhitasari Suratman

    2010-06-01

    Full Text Available Electropolymerization processes of pyrrole and the usage of polypyrrole membrane as lysine and alanine deliverance have been studied by cyclic voltammetry technique. Polypyrrole membrane was prepared by electropolymerization processes of pyrrole in water based solvent containing sodium perchlorate as supporting electrolyte. Electropolymerization processes were carried out within potential range of 0-1100 mV vs Ag/AgCl reference electrode and at the scanning rate of 100 mV/s. In this study, lysine and alanine have been used as molecules which could easily be loaded on and released from polypyrrole membrane. The presence of lysine or alanine during electropolymerization process reduced the rate of electropolymerization of polypyrrole. In lysine or alanine transfer processes into polypyrrole membrane, the interaction between polypyrrole and lysine or alanine showed by the curve of E½ oxidation in respect of - log C. It proved that the E½ oxidation shifted to more positive potential showed by the increasing of concentration of lysine or alanine. Beside that, voltammetric responses of lysine and alanine transfered into polypyrrole membrane were found to be Nernstian. The results indicated that polypyrrole could be used as a sensor of lysine and alanine.   Keywords: Electropolymerization, polypyrrole membrane, voltammetry technique

  20. Digestible lysine levels in diets supplemented with ractopamine

    Directory of Open Access Journals (Sweden)

    Evelar de Oliveira Souza

    2011-10-01

    Full Text Available In order evaluate digestible lysine levels in diets supplemented with 20 ppm of ractopamine on the performance and carcass traits, 64 barrows with high genetic potential at finishing phase were allotted in a completely randomized block design with four digestible lysine levels (0.80, 0.90, 1.00, and 1.10%, eight replicates and two pigs per experimental unit. Initial body weight and pigs' kinship were used as criteria in the blocks formation. Diets were mainly composed of corn and soybean meal supplemented with minerals, vitamins and amino acids to meet pigs' nutritional requirements at the finishing phase, except for digestible lysine. No effect of digestible lysine levels was observed in animal performance. The digestible lysine intake increased linearly by increasing the levels of digestible lysine in the diets. Carcass traits were not influenced by the dietary levels of digestible lysine. The level of 0.80% of digestible lysine in diets supplemented with 20 ppm ractopamine meets the nutritional requirements of castrated male pigs during the finishing phase.

  1. The Tale of Protein Lysine Acetylation in the Cytoplasm

    Directory of Open Access Journals (Sweden)

    Karin Sadoul

    2011-01-01

    Full Text Available Reversible posttranslational modification of internal lysines in many cellular or viral proteins is now emerging as part of critical signalling processes controlling a variety of cellular functions beyond chromatin and transcription. This paper aims at demonstrating the role of lysine acetylation in the cytoplasm driving and coordinating key events such as cytoskeleton dynamics, intracellular trafficking, vesicle fusion, metabolism, and stress response.

  2. Bioavailability of lysine in heat-treated foods and feedstuffs

    NARCIS (Netherlands)

    McArtney Rutherfurd, S.

    2010-01-01

    During the processing of foodstuffs, lysine can react with other compounds present to form nutritionally unavailable derivatives, the most common example of which are Maillard products. Maillard products can cause serious problems when determining the available lysine content of processed foods or f

  3. Bioavailability of lysine in heat-treated foods and feedstuffs

    NARCIS (Netherlands)

    McArtney Rutherfurd, S.

    2010-01-01

    During the processing of foodstuffs, lysine can react with other compounds present to form nutritionally unavailable derivatives, the most common example of which are Maillard products. Maillard products can cause serious problems when determining the available lysine content of processed foods or

  4. Tropical Tropospheric Ozone (TTO) Maps from Nimbus 7 and Earth-Probe TOMS by the Modified-Residual Method. 1; Validation, Evaluation and Trends based on Atlantic Regional Time Series

    Science.gov (United States)

    Thompson, Anne M.; Hudson, Robert D.

    1998-01-01

    The well-known wave-one pattern seen in tropical total ozone [Shiotani, 1992; Ziemke et al., 1996, 1998] has been used to develop a modified-residual (MR) method for retrieving time-averaged stratospheric ozone and tropospheric ozone column amount from TOMS (Total Ozone Mapping Spectrometer) over the 14 complete calendar years of Nimbus 7 observations (1979-1992) and from TOMS on the Earth-Probe (1996-present) and ADEOS platforms (1996- 1997). Nine- to sixteen-day averaged tropical tropospheric ozone (TTO) maps, validated with ozonesondes, show a seasonality expected from dynamical and chemical influences. The maps may be viewed on a homepage: http://metosrv2.umd.edu/tropo. Stratospheric column ozone, which is also derived by the modified-residual method, compares well with sondes (to within 6-7 DU) and with stratospheric ozone column derived from other satellites (within 8-10 DU). Validation of the TTO time-series is presently limited to ozonesonde comparisons with Atlantic stations and sites on the adjacent continents (Ascension Island, Natal, Brazil; Brazzaville); for the sounding periods, TTO at all locations agrees with the sonde record to +/-7 DU. TTO time-series and the magnitude of the wave-one pattern show ENSO signals in the strongest El Nifio periods from 1979-1998. From 12degN and 12degS, zonally averaged tropospheric ozone shows no significant trend from 1980-1990. Trends are also not significant during this period in localized regions, e.g. from just west of South America across to southern Africa. This is consistent with the ozonesonde record at Natal, Brazil (the only tropical ozone data publicly available for the 1980's), which shows a not statistically significant increase. The lack of trend in tropospheric ozone agrees with a statistical analysis based on another method for deriving TTO from TOMS, the so-called Convective-Cloud-Differential approach of Ziemke et al. [1998].

  5. Creative lysins: Listeria and the engineering of antimicrobial enzymes.

    Science.gov (United States)

    Van Tassell, Maxwell L; Angela Daum, M; Kim, Jun-Seob; Miller, Michael J

    2016-02-01

    Cell wall lytic enzymes have been of increasing interest as antimicrobials for targeting Gram-positive spoilage and pathogenic bacteria, largely due to the development of strains resistant to antibiotics and bacteriophage therapy. Such lysins show considerable promise against Listeria monocytogenes, a primary concern in food-processing environments, but there is room for improvement via protein engineering. Advances in antilisterial applications could benefit from recent developments in lysin biotechnology that have largely targeted other organisms. Herein we present various considerations for the future development of lysins, including environmental factors, cell physiology concerns, and dynamics of protein architecture. Our goal is to review key developments in lysin biotechnology to provide a contextual framework for the current models of lysin-cell interactions and highlight key considerations for the characterization and design of novel lytic enzymes.

  6. Cowpea mosaic virus VPg: sequencing of radiochemically modified protein allows mapping of the gene on B RNA

    Science.gov (United States)

    Zabel, Pim; Moerman, Marja; Lomonossoff, George; Shanks, Michael; Beyreuther, Konrad

    1984-01-01

    A partial amino acid sequence of cowpea mosaic virus (CPMV) VPg radiochemically modified by chloramine-T and Bolton-Hunter reagent has been determined. VPg covalently bound to viral RNA chains (VPg-RNA) was iodinated with chloramine-T and Bolton-Hunter reagent to label tyrosine and lysine residues, respectively. [125I]VPg-RNA was digested with nuclease P1 and the resulting [125I]VPg-pU was purified by SDS-polyacrylamide gel electrophoresis and subjected to automated Edman degradation. Control experiments with chemically synthesized poliovirus VPg showed the feasibility of radiochemical microsequence analysis of protein that had been radiochemically modified by chloramine-T and Bolton-Hunter reagent. Analysis of CPMV [125I]VPg-pU revealed the presence of tyrosine residues at position 12 and 14, and of lysine residues at position 3 and 20, respectively. In combination with Edman degradation of unlabeled CPMV VPg, which showed serine and arginine residues to be present at position 1 and 2, respectively, the data obtained allow the precise positioning of VPg within the 200 000 dalton (200 K) polyprotein encoded by CPMV B RNA and the prediction of its entire amino acid sequence. VPg is located at the COOH terminus of its 60 K, membrane-bound,precursor and proximal to the amino terminus of the protease-polymerase domain of the polyprotein. A processing scheme for the 200 K polyprotein is discussed in which Gln-Ser amino acid pairs act as the major signal for proteolytic cleavage. PMID:16453534

  7. Residue determination of glufosinate in plant origin foods using modified Quick Polar Pesticides (QuPPe) method and liquid chromatography coupled with tandem mass spectrometry.

    Science.gov (United States)

    Han, Yongtao; Song, Le; Zhao, Pengyue; Li, Yanjie; Zou, Nan; Qin, Yuhong; Li, Xuesheng; Pan, Canping

    2016-04-15

    A sensitive and specific method for the determination of glufosinate in plant origin foods was developed. The method involves extraction using modified QuPPe method, clean-up by multi-walled carbon nanotubes (MWCNTs), derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) and detection with liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The method was validated on twelve matrices spiked at 10 or 20, 100 and 500 μg/kg. The recovery ranged from 80% to 108% with intra-day RSDs (n=5) of 0.6-9.8% and inter-day RSDs (n=15) of 3.0-9.4%. Good linearities (R(2)⩾0.9991) were obtained for all matrices. The limit of detection (LOD) and limit of quantification (LOQ) for the selected matrices ranged from 0.3 to 3.3 μg kg(-1) and from 1 to 10 μg kg(-1), respectively. The method was demonstrated to be reliable and sensitive for the routine monitoring of glufosinate in plant origin foods.

  8. Residuation theory

    CERN Document Server

    Blyth, T S; Sneddon, I N; Stark, M

    1972-01-01

    Residuation Theory aims to contribute to literature in the field of ordered algebraic structures, especially on the subject of residual mappings. The book is divided into three chapters. Chapter 1 focuses on ordered sets; directed sets; semilattices; lattices; and complete lattices. Chapter 2 tackles Baer rings; Baer semigroups; Foulis semigroups; residual mappings; the notion of involution; and Boolean algebras. Chapter 3 covers residuated groupoids and semigroups; group homomorphic and isotone homomorphic Boolean images of ordered semigroups; Dubreil-Jacotin and Brouwer semigroups; and loli

  9. Enhanced Antimicrobial Activity of AamAP1-Lysine, a Novel Synthetic Peptide Analog Derived from the Scorpion Venom Peptide AamAP1

    Directory of Open Access Journals (Sweden)

    Ammar Almaaytah

    2014-04-01

    Full Text Available There is great interest in the development of antimicrobial peptides as a potentially novel class of antimicrobial agents. Several structural determinants are responsible for the antimicrobial and cytolytic activity of antimicrobial peptides. In our study, a new synthetic peptide analog, AamAP1-Lysine from the naturally occurring scorpion venom antimicrobial peptide AamAP1, was designed by modifying the parent peptide in order to increase the positive charge and optimize other physico-chemical parameters involved in antimicrobial activity. AamAP1-Lysine displayed potent antibacterial activity against Gram-positive and Gram-negative bacteria. The minimum inhibitory concentration was in the range of 5 to 15 µM with a 10 fold increase in potency over the parent peptide. The hemolytic and antiproliferative activity of AamAP1-Lysine against eukaryotic mammalian cells was minimal at the concentration range needed to inhibit bacterial growth. The antibacterial mechanism analysis indicated that AamAP1-Lysine is probably inducing bacterial cell death through membrane damage and permeabilization determined by the release of β-galactosidase enzyme from peptide treated E. coli cells. DNA binding studies revealed that AamAP1-Lysine caused complete retardation of DNA migration and could display intracellular activities in addition to the membrane permeabilization mode of action reported earlier. In conclusion, AamAP1-Lysine could prove to be a potential candidate for antimicrobial drug development in future studies.

  10. PLASTIC WASTE CONVERSION TO LIQUID FUELS OVER MODIFIED-RESIDUAL CATALYTIC CRACKING CATALYSTS: MODELING AND OPTIMIZATION USING HYBRID ARTIFICIAL NEURAL NETWORK – GENETIC ALGORITHM

    Directory of Open Access Journals (Sweden)

    Istadi Istadi

    2012-04-01

    Full Text Available The plastic waste utilization can be addressed toward different valuable products. A promising technology for the utilization is by converting it to fuels. Simultaneous modeling and optimization representing effect of reactor temperature, catalyst calcinations temperature, and plastic/catalyst weight ratio toward performance of liquid fuel production was studied over modified catalyst waste. The optimization was performed to find optimal operating conditions (reactor temperature, catalyst calcination temperature, and plastic/catalyst weight ratio that maximize the liquid fuel product. A Hybrid Artificial Neural Network-Genetic Algorithm (ANN-GA method was used for the modeling and optimization, respectively. The variable interaction between the reactor temperature, catalyst calcination temperature, as well as plastic/catalyst ratio is presented in surface plots. From the GC-MS characterization, the liquid fuels product was mainly composed of C4 to C13 hydrocarbons.KONVERSI LIMBAH PLASTIK MENJADI BAHAN BAKAR CAIR DENGAN METODE PERENGKAHAN KATALITIK MENGGUNAKAN KATALIS BEKAS YANG TERMODIFIKASI: PEMODELAN DAN OPTIMASI MENGGUNAKAN GABUNGAN METODE ARTIFICIAL NEURAL NETWORK DAN GENETIC ALGORITHM. Pemanfaatan limbah plastik dapat dilakukan untuk menghasilkan produk yang lebih bernilai tinggi. Salah satu teknologi yang menjanjikan adalah dengan mengkonversikannya menjadi bahan bakar. Permodelan, simulasi dan optimisasi simultan yang menggambarkan efek dari suhu reaktor, suhu kalsinasi katalis, dan rasio berat plastik/katalis terhadap kinerja produksi bahan bakar cair telah dipelajari menggunakan katalis bekas termodifikasi Optimisasi ini ditujukan untuk mencari kondisi operasi optimum (suhu reaktor, suhu kalsinasi katalis, dan rasio berat plastik/katalis yang memaksimalkan produk bahan bakar cair. Metode Hybrid Artificial Neural Network-Genetic Algorithm (ANN-GA telah digunakan untuk permodelan dan optimisasi simultan tersebut. Inetraksi antar variabel

  11. Coarse-grained simulations of hemolytic peptide δ-lysin interacting with a POPC bilayer.

    Science.gov (United States)

    King, Mariah J; Bennett, Ashley L; Almeida, Paulo F; Lee, Hee-Seung

    2016-12-01

    δ-lysin, secreted by a Gram-positive bacterium Staphylococcus aureus, is a 26-residue membrane active peptide that shares many common features with antimicrobial peptides (AMPs). However, it possesses a few unique features that differentiate itself from typical AMPs. In particular, δ-lysin has zero net charge, even though it has many charged residues, and it preferentially lyses eukaryotic cells over bacterial cells. Here, we present the results of coarse-grained molecular dynamics simulations of δ-lysin interacting with a zwitterionic membrane over a wide range of peptide concentrations. When the peptides concentration is low, spontaneous dimerization of peptides is observed on the membrane surface, but deep insertion of peptides or pore formation was not observed. However, the calculated free energy of peptide insertion suggests that a small fraction of peptides is likely to be present inside the membrane at the peptide concentrations typically seen in dye efflux experiments. When the simulations with multiple peptides are carried out with a single pre-inserted transmembrane peptide, spontaneous pore formation occurs with a peptide-to-lipid ratio (P/L) as low as P/L=1:42. Inter-peptide salt bridges among the transmembrane peptides seem to play a role in creating compact pores with very low level of hydration. More importantly, the transmembrane peptides making up the pore are constantly pushed to the opposite side of the membrane when the mass imbalance between the two sides of membrane is significant. Thus, the pore is very dynamic, allowing multiple peptides to translocate across the membrane simultaneously.

  12. 改性贵州玄武岩残积土的抗压强度试验研究%Experimental Research on Compressive Strength of Modified Residual Soil of Basalt in Guizhou

    Institute of Scientific and Technical Information of China (English)

    张瑞敏; 柴寿喜; 魏厚振; 徐良

    2012-01-01

    In order to make residual soil of basalt in guizhou underlying for subgrade reasonably, according to the different content of lime, fly ash and cement on the mixed with a single, double, orthogonal. Measuring the compressive strength on considering not water and water to get a best proportion . Test results show that: (1) The compressive strength of modified residual soil of basalt increase gradually with the increase of the content of modified materials with a single experiment. The modification treatment of lime soil reach a maximum 8% when is watered. Fly ash reach a maximum 15% when is not watered, at the same time, the watering sample are all crumbling. (2) The compressive strength increase all with the increase of the content with the double experiment, and the compressive strength of lime: fly ash = 1 : 1 is higher than lime = fly ash = 1 : 2. (3) The best proportion is for 8% lime, 8% fly ash, 2% cement with the orthogonal experiment, and the lime is the biggest influence factors to the compressive strength of residual soil. Because the compression strength all has different increases with lime, fly ash, cement three kinds of materials processing basalt residual soil, so it is reference for subgrade with three kinds of materials mixed processing basalt residual soil.%为了使贵州玄武岩残积土合理用于路基,通过采用石灰、粉煤灰、水泥三种改性材料按不同含量对其进行单掺、双掺、正交试验研究,同时考虑未浸水与浸水两种状态,测其抗压强度,得出最佳配比.实验结果表明:①单掺试验,改性残积土的抗压强度随着改性材料含量的增加而逐渐增大.石灰处理的改性土浸水时在8%达到最大值,粉煤灰处理的在未浸水时在15%达到最大值,同时浸水的试样全部崩解.②双掺试验,抗压强度均是随着含量的增加而增大,且石灰:粉煤灰=1∶1的抗压强度比石灰:粉煤灰=1∶2的高.③正交掺试验,得出

  13. Lysine Acetylation and Deacetylation in Brain Development and Neuropathies

    Directory of Open Access Journals (Sweden)

    Alicia Tapias

    2017-02-01

    Full Text Available Embryonic development is critical for the final functionality and maintenance of the adult brain. Brain development is tightly regulated by intracellular and extracellular signaling. Lysine acetylation and deacetylation are posttranslational modifications that are able to link extracellular signals to intracellular responses. A wealth of evidence indicates that lysine acetylation and deacetylation are critical for brain development and functionality. Indeed, mutations of the enzymes and cofactors responsible for these processes are often associated with neurodevelopmental and psychiatric disorders. Lysine acetylation and deacetylation are involved in all levels of brain development, starting from neuroprogenitor survival and proliferation, cell fate decisions, neuronal maturation, migration, and synaptogenesis, as well as differentiation and maturation of astrocytes and oligodendrocytes, to the establishment of neuronal circuits. Hence, fluctuations in the balance between lysine acetylation and deacetylation contribute to the final shape and performance of the brain. In this review, we summarize the current basic knowledge on the specific roles of lysine acetyltransferase (KAT and lysine deacetylase (KDAC complexes in brain development and the different neurodevelopmental disorders that are associated with dysfunctional lysine (deacetylation machineries.

  14. Mutation of aspartic acid-351, lysine-352, and lysine-515 alters the Ca2+ transport activity of the Ca2+-ATPase expressed in COS-1 cells.

    Science.gov (United States)

    Maruyama, K; MacLennan, D H

    1988-01-01

    Full-length cDNAs encoding neonatal and adult isoforms of the Ca2+-ATPase of rabbit fast-twitch skeletal muscle sarcoplasmic reticulum were expressed transiently in COS-1 cells. The microsomal fraction isolated from transfected COS-1 cells contained immunoreactive Ca2+-ATPase and catalyzed Ca2+ transport at rates at least 15-fold above controls. No differences were observed in either the rates or Ca2+ dependency of Ca2+ transport catalyzed by the two isoforms. Aspartic acid-351, the site of formation of the catalytic acyl phosphate in the enzyme, was mutated to asparagine, glutamic acid, serine, threonine, histidine, or alanine. In every case, Ca2+ transport activity and Ca2+-dependent phosphorylation were eliminated. Ca2+ transport was also eliminated by mutation of lysine-352 to arginine, glutamine, or glutamic acid or by mutation of Asp351-Lys352 to Lys351-Asp352. Mutation of lysine-515, the site of fluorescein isothiocyanate modification in the enzyme, resulted in diminished Ca2+ transport activity as follows: arginine, 60%; glutamine, 25%; glutamic acid, 5%. These results demonstrate the absolute requirement of acylphosphate formation for the Ca2+ transport function and define a residue important for ATP binding. They also demonstrate the feasibility of a thorough analysis of active sites in the Ca2+-ATPase by expression and site-specific mutagenesis. Images PMID:2966962

  15. HDAC inhibitors induce global changes in histone lysine and arginine methylation and alter expression of lysine demethylases.

    Science.gov (United States)

    Lillico, Ryan; Sobral, Marina Gomez; Stesco, Nicholas; Lakowski, Ted M

    2016-02-01

    Histone deacetylase (HDAC) inhibitors are cancer treatments that inhibit the removal of the epigenetic modification acetyllysine on histones, resulting in altered gene expression. Such changes in expression may influence other histone epigenetic modifications. We describe a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify lysine acetylation and methylation and arginine methylation on histones extracted from cultured cells treated with HDAC inhibitors. The HDAC inhibitors vorinostat, mocetinostat and entinostat induced 400-600% hyperacetylation in HEK 293 and K562 cells. All HDAC inhibitors decreased histone methylarginines in HEK 293 cells but entinostat produced dose dependent reductions in asymmetric dimethylarginine, not observed in K562 cells. Vorinostat produced increases in histone lysine methylation and decreased expression of some lysine demethylases (KDM), measured by quantitative PCR. Entinostat had variable effects on lysine methylation and decreased expression of some KDM while increasing expression of others. Mocetinostat produced dose dependent increases in histone lysine methylation by LC-MS/MS. This was corroborated with a multiplex colorimetric assay showing increases in histone H3 lysine 4, 9, 27, 36 and 79 methylation. Increases in lysine methylation were correlated with dose dependent decreases in the expression of seven KDM. Mocetinostat functions as an HDAC inhibitor and a de facto KDM inhibitor.

  16. Design and construction of novel molecular conjugates for signal amplification (I): conjugation of multiple horseradish peroxidase molecules to immunoglobulin via primary amines on lysine peptide chains.

    Science.gov (United States)

    Dhawan, Subhash

    2002-12-01

    Immunoconjugates are widely used for indirect detection of analytes (such as antibodies or antigens) in a variety of immunoassays. However, the availability of functional groups such as primary amines or free sulfhydryls in an immunoglobulin molecule is the limiting factor for optimal conjugation and, therefore, determines the sensitivity of an assay. In the present study, an N-terminal bromoacetylated 20 amino acid peptide containing 20 lysine residues was conjugated to N-succinimidyl-S-acetylthioacetate (SATA)-modified IgG or free sulfhydryl groups on 2-mercaptoethylamine (2-MEA)-reduced IgG molecules via a thioether (S[bond]CH(2)CONH) linkage to introduce multiple reactive primary amines per IgG. These primary amines were then covalently coupled with maleimide-activated horseradish peroxidase (HRP). The poly-HRP-antibody conjugates thus generated demonstrated greater than 15-fold signal amplification upon reaction with orthophenyldiamine substrate. The poly-HRP-antibody conjugates efficiently detected human immunodeficiency virus (HIV)-1 antibodies in plasma specimens with significantly higher sensitivity than conventionally prepared HRP-antibody conjugates in an HIV-1 solid-phase enzyme immunoassay and Western blot analysis. The signal amplification techniques reported here could have the potential for development of highly sensitive immunodiagnostic assay systems.

  17. Tropical tropospheric ozone (TTO) maps from Nimbus 7 and Earth Probe TOMS by the modified-residual method: Evaluation with sondes, ENSO signals, and trends from Atlantic regional time series

    Science.gov (United States)

    Thompson, Anne M.; Hudson, Robert D.

    1999-11-01

    The modified-residual (MR) method for retrieving time-averaged stratospheric ozone and tropospheric ozone column amounts from the Total Ozone Mapping Spectrometer (TOMS) is applied to the 14 complete calendar years of Nimbus 7 observations (1979-1992). These are available as digital data at http://metosrv2.umd.edu/˜tropo/14y_data.d The MR method has also been used to produce real-time maps of tropical tropospheric ozone (TTO) from TOMS on the Earth-Probe (1996-present) and ADEOS platforms (1996-1997). Evaluation of the TTO time series for 1979-1990 and 1997-1998 is presented here; it is limited to the few tropical ozonesonde stations operational during those years (Ascension Island; Natal, Brazil; Brazzaville). The standard deviation of the differences between TTO and the sondes is ±(6-7) Dobson units (DU), depending on location. Stratospheric column ozone, which is also derived by the modified-residual method, compares favorably with sondes (to within 6-9 DU) and with stratospheric ozone inferred from other satellites (usually 8-15 DU lower than the latter). TTO time series and the magnitude of the tropospheric wave-one pattern show El Niño-Southern Oscillation (ENSO) signals during the period from 1979-1992. During 1997 the ENSO stands out at some stations, but not at others. Between 12°N and 12°S, zonally averaged TTO shows no significant trend from 1980-1990. Trends are also not significant during this period in localized regions, for example, from just west of South America across to southern Africa. This is consistent with the ozonesonde record at Natal, Brazil (the only tropical ozone data publicly available for the 1980s), which shows no significant trend. The lack of trend in tropospheric ozone agrees with a statistical analysis based on another method for deriving TTO from TOMS, the convective-cloud-differential approach of Ziemke et al. [1998].

  18. Duplicate abalone egg coat proteins bind sperm lysin similarly, but evolve oppositely, consistent with molecular mimicry at fertilization.

    Directory of Open Access Journals (Sweden)

    Jan E Aagaard

    Full Text Available Sperm and egg proteins constitute a remarkable paradigm in evolutionary biology: despite their fundamental role in mediating fertilization (suggesting stasis, some of these molecules are among the most rapidly evolving ones known, and their divergence can lead to reproductive isolation. Because of strong selection to maintain function among interbreeding individuals, interacting fertilization proteins should also exhibit a strong signal of correlated divergence among closely related species. We use evidence of such molecular co-evolution to target biochemical studies of fertilization in North Pacific abalone (Haliotis spp., a model system of reproductive protein evolution. We test the evolutionary rates (d(N/d(S of abalone sperm lysin and two duplicated egg coat proteins (VERL and VEZP14, and find a signal of co-evolution specific to ZP-N, a putative sperm binding motif previously identified by homology modeling. Positively selected residues in VERL and VEZP14 occur on the same face of the structural model, suggesting a common mode of interaction with sperm lysin. We test this computational prediction biochemically, confirming that the ZP-N motif is sufficient to bind lysin and that the affinities of VERL and VEZP14 are comparable. However, we also find that on phylogenetic lineages where lysin and VERL evolve rapidly, VEZP14 evolves slowly, and vice versa. We describe a model of sexual conflict that can recreate this pattern of anti-correlated evolution by assuming that VEZP14 acts as a VERL mimic, reducing the intensity of sexual conflict and slowing the co-evolution of lysin and VERL.

  19. protein, tryptophan and lysine contents in quality protien maize ...

    African Journals Online (AJOL)

    owner

    for human nutrition recommended by Food and Agriculture Organization in ... METHODS: The protein, tryptophan and lysine contents of improved ... This study revealed the fact that genetic factor influences the protein, ... Ethiop J Health Sci.

  20. Ethanol Fermentation Performance of Grain Sorghums (Sorghum bicolor) with Modified Endosperm Matrices

    Energy Technology Data Exchange (ETDEWEB)

    Wu, X.; Jampala, B; Robbins, A; Hays, D; Yan, S; Xu, F; Rooney, W; Peterson, G; Shi, Y; Wang, D

    2010-01-01

    We tested 13 sorghum entries (lines and hybrids) with different endosperm matrices for ethanol production using a laboratory dry grind process. Waxy and heterowaxy samples had the highest efficiencies. Free amino nitrogen (FAN) contents in sorghum samples were positively related to the fermentation rate during fermentation (R{sup 2} = 0.8618). Dried distiller's grain with solubles (DDGS) from different sorghums had significantly different crude protein and crude fat contents. Residual starch content in DDGS ranged from 0.60% for the most efficient sample to 2.66% for the least efficient sample. This study showed that the HD lines (TX1, TX3, TX5, TX7, and TX9) with modified endosperm protein matrix have several attributes desirable for ethanol production: easily pasted starch granules, significantly higher FAN content in finished mashes, 30-45% faster ethanol fermentation rate during early stages, and 50-60% higher lysine content in DDGS.

  1. Digestible lysine levels in diets for laying Japanese quails

    Directory of Open Access Journals (Sweden)

    Cleverson Luís Nascimento Ribeiro

    2013-07-01

    Full Text Available The objective of this study was to estimate the digestible lysine requirement of Japanese quails in the egg-laying phase. A total of 336 female Japanese quails (Coturnix coturnix japonica of average initial age of 207 days were distributed in a completely randomized experimental design, composed of 6 treatments (lysine levels with 7 replicates and 8 birds per experimental unit, with duration of 84 days. Experimental diets were formulated from a basal diet, with corn and soybean meal, with 2.800 kcal ME/kg and 203.70 g/kg crude protein, showing levels of 9.50; 10.00; 10.50; 11.00; 11.50; and 12.00 g/kg digestible lysine; diets remained isoprotein and isocaloric. The following variables were studied: feed intake (FI; lysine intake (LI; egg production per bird per day (EPBD; egg production per bird housed (EPBH; production of marketable eggs (PME; egg weight (EW; egg mass (EM; utilization efficiency of lysine for egg mass production (UELEM; feed conversion per mass (FCEM; feed conversion per dozen eggs (FCDZ; bird availability (BA; percentages of yolk (Y, albumen (A and shell (S; specific egg weight (SW; nitrogen ingested (NI; nitrogen excreted (NE; and nitrogen balance (NB. Significant effect was only observed for LI, EW, EM, UELEM, FCEM, Y, A and SW. The digestible lysine level estimated in diets for laying Japanese quails is 11.20 g digestible lysine/kg diet, corresponding to an average daily intake of 272.23 mg lysine.

  2. H3 lysine 4 is acetylated at active gene promoters and is regulated by H3 lysine 4 methylation.

    Directory of Open Access Journals (Sweden)

    Benoit Guillemette

    2011-03-01

    Full Text Available Methylation of histone H3 lysine 4 (H3K4me is an evolutionarily conserved modification whose role in the regulation of gene expression has been extensively studied. In contrast, the function of H3K4 acetylation (H3K4ac has received little attention because of a lack of tools to separate its function from that of H3K4me. Here we show that, in addition to being methylated, H3K4 is also acetylated in budding yeast. Genetic studies reveal that the histone acetyltransferases (HATs Gcn5 and Rtt109 contribute to H3K4 acetylation in vivo. Whilst removal of H3K4ac from euchromatin mainly requires the histone deacetylase (HDAC Hst1, Sir2 is needed for H3K4 deacetylation in heterochomatin. Using genome-wide chromatin immunoprecipitation (ChIP, we show that H3K4ac is enriched at promoters of actively transcribed genes and located just upstream of H3K4 tri-methylation (H3K4me3, a pattern that has been conserved in human cells. We find that the Set1-containing complex (COMPASS, which promotes H3K4me2 and -me3, also serves to limit the abundance of H3K4ac at gene promoters. In addition, we identify a group of genes that have high levels of H3K4ac in their promoters and are inadequately expressed in H3-K4R, but not in set1Δ mutant strains, suggesting that H3K4ac plays a positive role in transcription. Our results reveal a novel regulatory feature of promoter-proximal chromatin, involving mutually exclusive histone modifications of the same histone residue (H3K4ac and H3K4me.

  3. Prioritizing stream types according to their potential risk to receive crop plant material--A GIS-based procedure to assist in the risk assessment of genetically modified crops and systemic insecticide residues.

    Science.gov (United States)

    Bundschuh, Rebecca; Kuhn, Ulrike; Bundschuh, Mirco; Naegele, Caroline; Elsaesser, David; Schlechtriemen, Ulrich; Oehen, Bernadette; Hilbeck, Angelika; Otto, Mathias; Schulz, Ralf; Hofmann, Frieder

    2016-03-15

    Crop plant residues may enter aquatic ecosystems via wind deposition or surface runoff. In the case of genetically modified crops or crops treated with systemic pesticides, these materials may contain insecticidal Bt toxins or pesticides that potentially affect aquatic life. However, the particular exposure pattern of aquatic ecosystems (i.e., via plant material) is not properly reflected in current risk assessment schemes, which primarily focus on waterborne toxicity and not on plant material as the route of uptake. To assist in risk assessment, the present study proposes a prioritization procedure of stream types based on the freshwater network and crop-specific cultivation data using maize in Germany as a model system. To identify stream types with a high probability of receiving crop materials, we developed a formalized, criteria-based and thus transparent procedure that considers the exposure-related parameters, ecological status--an estimate of the diversity and potential vulnerability of local communities towards anthropogenic stress--and availability of uncontaminated reference sections. By applying the procedure to maize, ten stream types out of 38 are expected to be the most relevant if the ecological effects from plant-incorporated pesticides need to be evaluated. This information is an important first step to identifying habitats within these stream types with a high probability of receiving crop plant material at a more local scale, including accumulation areas. Moreover, the prioritization procedure developed in the present study may support the selection of aquatic species for ecotoxicological testing based on their probability of occurrence in stream types having a higher chance of exposure. Finally, this procedure can be adapted to any geographical region or crop of interest and is, therefore, a valuable tool for a site-specific risk assessment of crop plants carrying systemic pesticides or novel proteins, such as insecticidal Bt toxins, expressed

  4. Factors affecting protein transfer into surfactant-isooctane solution: a case study of extraction behavior of chemically modified cytochrome c.

    Science.gov (United States)

    Ono, T; Goto, M

    1998-01-01

    The extraction mechanism of proteins by surfactant molecules in an organic solvent has been investigated using a chemically modified protein. We conducted guanidylation on lysine residues of cytochrome c by replacing their amino groups with homoarginine to enhance the protein-surfactant interaction. Results have shown that guanidylated cytochrome c readily forms a hydrophobic complex with dioleyl phosphoric acid (DOLPA) through hydrogen bonding between the phosphate moiety and the guanidinium groups. Although improved protein-surfactant interaction activated the formation of a hydrophobic complex at the interface, it could not improve the protein transfer in isooctane. It has been established that the protein extraction mechanism using surfactant molecules is mainly governed by two processes: formation of an interfacial complex at the oil-water interface and the subsequent solubilization of the complex into the organic phase. In addition, a kinetic study demonstrated that guanidylation of lysine accelerated the initial extraction rate of cytochrome c. This fact implies that the protein transferability from aqueous phase into organic phase depends on the protein-surfactant interaction which can be modified by protein surface engineering.

  5. Maintenance requirement and deposition efficiency of lysine in pigs

    Directory of Open Access Journals (Sweden)

    Marcos Speroni Ceron

    2013-09-01

    Full Text Available The objective of this work was to determine the maintenance requirement and the deposition efficiency of lysine in growing pigs. It was used the incomplete changeover experimental design, with replicates over time. Twelve castrated pigs with average body weight (BW of 52±2 kg were kept in metabolism crates with a controlled temperature of 22ºC. The diets were formulated to supply 30, 50, 60, and 70% of the expected requirements of standardized lysine, and provided at 2.6 times the energy requirements for maintenance. The trial lasted 24 days and was divided into two periods of 12 days: seven days for animal adaptation to the diet and five days for sample collection. The increasing content of lysine in the diet did not affect dry matter intake of the pigs. The amount of nitrogen excreted was 47% of the nitrogen intake, of which 35% was excreted through feces and 65% through urine. The estimated endogenous losses of lysine were 36.4 mg kg-1 BW0.75. The maintenance requirement of lysine for pigs weighing around 50 kg is 40.4 mg kg-1 BW0.75, and the deposition efficiency of lysine is 90%.

  6. Targeting protein lysine methylation and demethylation in cancers

    Institute of Scientific and Technical Information of China (English)

    Yunlong He; Ilia Korboukh; Jian Jin; Jing Huang

    2012-01-01

    During the last decade,we saw an explosion of studies investigating the role of lysine methylation/demethylation of histones and non-histone proteins,such as p53,NF-kappaB,and E2F1.These ‘Ying-Yang' post-translational modifications are important to fine-tuning the activity of these proteins. Lysine methylation and demethylation are catalyzed by protein lysine methyltransferases (PKMTs) and protein lysine demethylases (PKDMs).PKMTs,PKDMs,and their substrates have been shown to play important roles in cancers.Although the underlying mechanisms of tumorigenesis are still largely unknown,growing evidence is starting to link aberrant regulation of methylation to tumorigenesis.This review focuses on summarizing the recent progress in understanding of the function of protein lysine methylation,and in the discovery of small molecule inhibitors for PKMTs and PKDMs.We also discuss the potential and the caveats of targeting protein lysine methylation for the treatment of cancer.

  7. Lysine-functionalized nanodiamonds: synthesis, physiochemical characterization, and nucleic acid binding studies

    Directory of Open Access Journals (Sweden)

    Kaur R

    2012-07-01

    . These modified NDs formed highly stable aqueous dispersions with a zeta potential of 49 mV and particle size of approximately 20 nm. The functionalized NDs were found to be able to bind plasmid DNA and small interfering RNA by forming nanosized "diamoplexes".Conclusion: The lysine-substituted ND particles generated in this study exhibit stable aqueous formulations and show potential for use as carriers for genetic materials.Keywords: disaggregation, spectroscopy, dispersion, electrophoresis, size, zeta potential

  8. Enzymic and chemical synthesis of epilson-N-(L-propionyl-2)-L-lysine.

    Science.gov (United States)

    Fujioka, M; Tanaka, M

    1978-10-01

    Pyruvate was shown to act as an oxo acid substrate in the reverse direction of saccharopine dehydrogenase [epsilon N-(L-glutaryl-2)-L-lysine: NAD oxidoreductase (L-lysine-forming)] reaction. The enzymic condensation product of lysine and pyruvate was isolated and identified as epsilon-N-(L-propionyl-2)-L-lysine by comparison with the synthetic compound. A method for the chemical preparation of diastereoisomers of epsilon-N-(propionyl-2)-L-lysine is also described.

  9. l-lysine production by Bacillus methanolicus: Genome-based mutational analysis and l-lysine secretion engineering.

    Science.gov (United States)

    Nærdal, Ingemar; Netzer, Roman; Irla, Marta; Krog, Anne; Heggeset, Tonje Marita Bjerkan; Wendisch, Volker F; Brautaset, Trygve

    2017-02-20

    Bacillus methanolicus is a methylotrophic bacterium with an increasing interest in academic research and for biotechnological applications. This bacterium was previously applied for methanol-based production of l-glutamate, l-lysine and the five-carbon diamine cadaverine by wild type, classical mutant and recombinant strains. The genomes of two different l-lysine secreting B. methanolicus classical mutant strains, NOA2#13A52-8A66 and M168-20, were sequenced. We focused on mutational mapping in genes present in l-lysine and other relevant amino acid biosynthetic pathways, as well as in the primary cell metabolism important for precursor supply. In addition to mutations in the aspartate pathway genes dapG, lysA and hom-1, new mutational target genes like alr, proA, proB1, leuC, odhA and pdhD were identified. Surprisingly, no mutations were found in the putative l-lysine transporter gene lysE(MGA3). Inspection of the wild type B. methanolicus strain PB1 genome sequence identified two homologous putative l-lysine transporter genes, lysE(PB1) and lysE2(PB1). The biological role of these putative l-lysine transporter genes, together with the heterologous l-lysine exporter gene lysE(Cg) from Corynebacterium glutamicum, were therefore investigated. Our results demonstrated that the titer of secreted l-lysine in B. methanolicus was significantly increased by overexpression of lysE(Cg) while overexpression of lysE(MGA3), lysE(PB1) and lysE2(PB1) had no measurable effect.

  10. Affecting proton mobility in activated peptide and whole protein ions via lysine guanidination.

    Science.gov (United States)

    Pitteri, Sharon J; Reid, Gavin E; McLuckey, Scott A

    2004-01-01

    We have evaluated the effect of lysine guanidination in peptides and proteins on the dissociation of protonated ions in the gas phase. The dissociation of guanidinated model peptide ions compared to their unmodified forms showed behavior consistent with concepts of proton mobility as a major factor in determining favored fragmentation channels. Reduction of proton mobility associated with lysine guanidination was reflected by a relative increase in cleavages occurring C-terminal to aspartic acid residues as well as increases in small molecule losses. To evaluate the effect of guanidination on the dissociation behavior of whole protein ions, bovine ubiquitin was selected as a model. Essentially, all of the amide bond cleavages associated with the +10 charge state of fully guanidinated ubiquitin were observed to occur C-terminal to aspartic acid residues, unlike the dissociation behavior of the +10 ion of the unmodified protein, where competing cleavage N-terminal to proline and nonspecific amide bond cleavages were also observed. The +8 and lower charge states of the guanidinated protein showed prominent losses of small neutral molecules. This overall fragmentation behavior is consistent with current hypotheses regarding whole protein dissociation that consider proton mobility and intramolecular charge solvation as important factors in determining favored dissociation channels, and are also consistent with the fragmentation behaviors observed for the guanidinated model peptide ions. Further evaluation of the utility of condensed phase guanidination of whole proteins is necessary but the results described here confirm that guanidination can be an effective strategy for enhancing C-terminal aspartic acid cleavages. Gas phase dissociation exclusively at aspartic acid residues, especially for whole protein ions, could be useful in identifying and characterizing proteins via tandem mass spectrometry of whole protein ions.

  11. The strong anti-glioblastoma capacity of the plasma-stimulated lysine-rich medium

    Science.gov (United States)

    Yan, Dayun; Nourmohammadi, Niki; Talbot, Annie; Sherman, Jonathan H.; Keidar, Michael

    2016-07-01

    Plasma-stimulated medium (PSM) shows a remarkable anti-cancer capacity as strong as the direct cold atmospheric plasma (CAP) treatment of cancer cells. PSM is able to effectively resist the growth of several cancer cell lines. To date, the sole approach to strengthen the anti-cancer capacity of PSM is extending the plasma treatment time. In this study, we demonstrated that the anti-glioblastoma capacity of PSM could be significantly increased by adding 20 mM lysine in Dulbecco’s modified Eagle’s medium (DMEM). This study provides clear evidence that the anti-glioblastoma capacity of PSM could be noticeably enhanced by modifying the composition of medium without increasing the CAP treatment time.

  12. Lysine acetylation stoichiometry and proteomics analyses reveal pathways regulated by sirtuin 1 in human cells.

    Science.gov (United States)

    Gil, Jeovanis; Ramírez-Torres, Alberto; Chiappe, Diego; Luna-Peñaloza, Juan; Fernandez-Reyes, Francis C; Arcos-Encarnación, Bolivar; Contreras, Sandra; Encarnación-Guevara, Sergio

    2017-09-11

    Lysine acetylation is a widespread posttranslational modification (PTM) affecting many biological pathways. Recent studies indicate that acetylated lysine residues mainly exhibit low acetylation occupancy, but challenges in sample preparation and analysis make it difficult to confidently assign these numbers, limiting understanding of their biological significance. Here, we tested three common sample preparation methods to determine their suitability for assessing acetylation stoichiometry in three human cell lines, identifying the acetylation occupancy in more than 1,300 proteins from each cell line. The stoichiometric analysis in combination with quantitative proteomics also enabled us to explore their functional roles. We found that higher abundance of the deacetylase sirtuin 1 (SIRT1) correlated with lower acetylation occupancy and lower levels of ribosomal proteins including those involved in ribosome biogenesis and rRNA processing. Treatment with the SIRT1 inhibitor EX-527 confirmed SIRT1's role in the regulation of pre-rRNA synthesis and processing. Specifically, proteins involved in pre-rRNA transcription, including subunits of the Pol 1 and SL1 complexes and the RNA polymerase I specific transcription initiation factor RRN3 were up-regulated after SIRT1 inhibition. Moreover, many protein effectors and regulators of pre-rRNA processing needed for rRNA maturation were also up-regulated after EX-527 treatment, with the outcome that pre-rRNA and 28S rRNA levels also increased. More generally, we found that SIRT1 inhibition down-regulates metabolic pathways including glycolysis and pyruvate metabolism. Together, these results provide the largest dataset thus far of lysine acetylation stoichiometry (available via ProteomeXchange with identifier PXD005903) and set the stage for further biological investigations of this central PTM. Copyright © 2017, The American Society for Biochemistry and Molecular Biology.

  13. Converting the yeast arginine can1 permease to a lysine permease.

    Science.gov (United States)

    Ghaddar, Kassem; Krammer, Eva-Maria; Mihajlovic, Natalija; Brohée, Sylvain; André, Bruno; Prévost, Martine

    2014-03-01

    Amino acid uptake in yeast cells is mediated by about 16 plasma membrane permeases, most of which belong to the amino acid-polyamine-organocation (APC) transporter family. These proteins display various substrate specificity ranges. For instance, the general amino acid permease Gap1 transports all amino acids, whereas Can1 and Lyp1 catalyze specific uptake of arginine and lysine, respectively. Although Can1 and Lyp1 have different narrow substrate specificities, they are close homologs. Here we investigated the molecular rules determining the substrate specificity of the H(+)-driven arginine-specific permease Can1. Using a Can1-Lyp1 sequence alignment as a guideline and a three-dimensional Can1 structural model based on the crystal structure of the bacterial APC family arginine/agmatine antiporter, we introduced amino acid substitutions liable to alter Can1 substrate specificity. We show that the single substitution T456S results in a Can1 variant transporting lysine in addition to arginine and that the combined substitutions T456S and S176N convert Can1 to a Lyp1-like permease. Replacement of a highly conserved glutamate in the Can1 binding site leads to variants (E184Q and E184A) incapable of any amino acid transport, pointing to a potential role for this glutamate in H(+) coupling. Measurements of the kinetic parameters of arginine and lysine uptake by the wild-type and mutant Can1 permeases, together with docking calculations for each amino acid in their binding site, suggest a model in which residues at positions 176 and 456 confer substrate selectivity at the ligand-binding stage and/or in the course of conformational changes required for transport.

  14. Click modification of helical amylose by poly(L-lysine) dendrons for non-viral gene delivery

    Energy Technology Data Exchange (ETDEWEB)

    Pang, Jia-Dong [PCFM Lab and GDHPPC Lab, Institute of Polymer Science, Department of Polymer and Materials Science, School of Chemistry and Chemical Engineering, Sun Yat-sen (Zhongshan) University, Guangzhou 510275 (China); Zhuang, Bao-Xiong [Second Affiliated Hospital, Sun Yat-sen (Zhongshan) University, Guangzhou 510102 (China); Mai, Kaijin [PCFM Lab and GDHPPC Lab, Institute of Polymer Science, Department of Polymer and Materials Science, School of Chemistry and Chemical Engineering, Sun Yat-sen (Zhongshan) University, Guangzhou 510275 (China); Chen, Ru-Fu [Second Affiliated Hospital, Sun Yat-sen (Zhongshan) University, Guangzhou 510102 (China); Wang, Jie, E-mail: sumsjw@163.com [Second Affiliated Hospital, Sun Yat-sen (Zhongshan) University, Guangzhou 510102 (China); Zhang, Li-Ming, E-mail: ceszhlm@mail.sysu.edu.cn [PCFM Lab and GDHPPC Lab, Institute of Polymer Science, Department of Polymer and Materials Science, School of Chemistry and Chemical Engineering, Sun Yat-sen (Zhongshan) University, Guangzhou 510275 (China)

    2015-04-01

    Although amylose as a naturally-occurring helical polysaccharide has been widely used for biomedical applications, few studies have dealt with its chemical modification for non-viral gene delivery. In this work, the click modification of amylose by poly(L-lysine) dendrons was carried out and then characterized by Fourier transform infrared spectroscopy, wide-angle X-ray diffraction and elemental analyses. Such a modified polysaccharide exhibited excellent ability to condense plasmid pMSCV-GFP-PARK2 into compact and spherical nanoparticles. Moreover, it displayed much lower cytotoxicity when compared to branched polyethylenimine (bPEI, 25 kDa), a commercially available gene vector. Similar to bPEI, it had a dose-dependent gene transfection activity in human embryonic kidney 293T cells, as observed by confocal laser scanning microscopy and flow cytometry. At each optimized N/P ratio, the percentage of transfected cells by this modified polysaccharide was found to be comparable to that by bPEI. Western blot and cell apoptosis analyses confirmed its effectiveness for the delivery of plasmid pMSCV-GFP-PARK2 to 293T cells. - Highlights: • The click modification of amylose by poly(L-lysine) dendrons was carried out. • This modified amylose could condense plasmid pMSCV-GFP-PARK2 into nanocomplexes. • This modified amylose exhibited much lower cytotoxicity than commercial polyethylenimine. • This modified amylose could delivery efficiently plasmid pMSCV-GFP-PARK2 to 293T cells.

  15. Crystal Structure of the Lysine Riboswitch Regulatory mRNA Element

    Energy Technology Data Exchange (ETDEWEB)

    Garst, A.; Heroux, A; Rambo, R; Batey, R

    2008-01-01

    Riboswitches are metabolite-sensitive elements found in mRNAs that control gene expression through a regulatory secondary structural switch. Along with regulation of lysine biosynthetic genes, mutations within the lysine-responsive riboswitch (L-box) play a role in the acquisition of resistance to antimicrobial lysine analogs. To understand the structural basis for lysine binding, we have determined the 2.8{angstrom} resolution crystal structure of lysine bound to the Thermotoga maritima asd lysine riboswitch ligand-binding domain. The structure reveals a complex architecture scaffolding a binding pocket completely enveloping lysine. Mutations conferring antimicrobial resistance cluster around this site as well as highly conserved long range interactions, indicating that they disrupt lysine binding or proper folding of the RNA. Comparison of the free and bound forms by x-ray crystallography, small angle x-ray scattering, and chemical probing reveals almost identical structures, indicating that lysine induces only limited and local conformational changes upon binding.

  16. Lysine methylation-dependent binding of 53BP1 to the pRb tumor suppressor.

    Science.gov (United States)

    Carr, Simon M; Munro, Shonagh; Zalmas, Lykourgos-Panagiotis; Fedorov, Oleg; Johansson, Catrine; Krojer, Tobias; Sagum, Cari A; Bedford, Mark T; Oppermann, Udo; La Thangue, Nicholas B

    2014-08-01

    The retinoblastoma tumor suppressor protein pRb is a key regulator of cell cycle progression and mediator of the DNA damage response. Lysine methylation at K810, which occurs within a critical Cdk phosphorylation motif, holds pRb in the hypophosphorylated growth-suppressing state. We show here that methyl K810 is read by the tandem tudor domain containing tumor protein p53 binding protein 1 (53BP1). Structural elucidation of 53BP1 in complex with a methylated K810 pRb peptide emphasized the role of the 53BP1 tandem tudor domain in recognition of the methylated lysine and surrounding residues. Significantly, binding of 53BP1 to methyl K810 occurs on E2 promoter binding factor target genes and allows pRb activity to be effectively integrated with the DNA damage response. Our results widen the repertoire of cellular targets for 53BP1 and suggest a previously unidentified role for 53BP1 in regulating pRb tumor suppressor activity.

  17. The putative oncogene GASC1 demethylates tri- and dimethylated lysine 9 on histone H3

    DEFF Research Database (Denmark)

    Cloos, Paul A C; Christensen, Jesper; Agger, Karl;

    2006-01-01

    Methylation of lysine and arginine residues on histone tails affects chromatin structure and gene transcription. Tri- and dimethylation of lysine 9 on histone H3 (H3K9me3/me2) is required for the binding of the repressive protein HP1 and is associated with heterochromatin formation...... and transcriptional repression in a variety of species. H3K9me3 has long been regarded as a 'permanent' epigenetic mark. In a search for proteins and complexes interacting with H3K9me3, we identified the protein GASC1 (gene amplified in squamous cell carcinoma 1), which belongs to the JMJD2 (jumonji domain containing...... 2) subfamily of the jumonji family, and is also known as JMJD2C. Here we show that three members of this subfamily of proteins demethylate H3K9me3/me2 in vitro through a hydroxylation reaction requiring iron and alpha-ketoglutarate as cofactors. Furthermore, we demonstrate that ectopic expression...

  18. Design and synthesis of benzodiazepine analogs as isoform-selective human lysine deacetylase inhibitors.

    Science.gov (United States)

    Reddy, D Rajasekhar; Ballante, Flavio; Zhou, Nancy J; Marshall, Garland R

    2017-02-15

    A comprehensive investigation was performed to identify new benzodiazepine (BZD) derivatives as potent and selective human lysine deacetylase inhibitors (hKDACis). A total of 108 BZD compounds were designed, synthesized and from that 104 compounds were biologically evaluated against human lysine deacetylases (hKDACs) 1, 3 and 8 (class I) and 6 (class IIb). The most active compounds showed mid-nanomolar potencies against hKDACs 1, 3 and 6 and micromolar activity against hKDAC8, while a promising compound (6q) showed selectivity towards hKDAC3 among the different enzyme isoforms. An hKDAC6 homology model, refined by molecular dynamics simulation was generated, and molecular docking studies performed to rationalize the dominant ligand-residue interactions as well as to define structure-activity-relationships. Experimental results confirmed the usefulness of the benzodiazepine moiety as capping group when pursuing hKDAC isoform-selectivity inhibition, suggesting its continued use when designing new hKDACis.

  19. Effects of lysine-induced acute renal failure in dogs.

    Science.gov (United States)

    Asanuma, Kentaro; Adachi, Kenji; Sugimoto, Tetsuro; Chiba, Shuichi

    2006-05-01

    This study investigates the effects of lysine-induced acute renal failure. Female dogs received a lysine hydrochloride (lysine) of 4500 mg/kg/day (3.75 ml/kg/hr) for 3 consecutive days. The dogs were observed for clinical signs. Body weights were recorded, food consumption and water consumption calculated, and urinalysis and blood biochemistry were performed daily. Plasma samples for amino acid determinations were obtained from all dogs, which were necropsied on Day 3. Histopathological examinations were done on all test animals. Compound-related findings include the following. Blood biochemistry results showed increases in ammonia, blood urea nitrogen, blood urea nitrogen/creatinine ratio, and creatinine. Urinary changes consisted of increases in urine volume, total protein, albumin, gamma-glutamyl transpeptidase, and N-acetyl-beta-D-glucosaminidase. In addition, macroscopic findings consisted of pale, congested capsule; microscopic findings consisted of hypertrophy of proximal convoluted tubule (mainly S1 segment), and degeneration/desquamation of urinary tubule (mainly S3 segment with hyaline casts) in the kidney. From these findings, it can be concluded that lysine is nephrotoxic in dogs. Nephrotoxicity of lysine may relate to direct tubular toxicity and to tubular obstruction.

  20. Antioxidant activity of carbocysteine lysine salt monohydrate.

    Science.gov (United States)

    Pinamonti, S; Venturoli, L; Leis, M; Chicca, M; Barbieri, A; Sostero, S; Ravenna, F; Daffonchio, L; Novellini, R; Ciaccia, A

    2001-09-01

    Reactive oxygen radicals are involved in many respiratory diseases, including chronic obstructive pulmonary disease (COPD). Carbocysteine lysine salt monohydrate (CLS) is a mucoactive drug effective in the treatment of bronchopulmonary diseases characterized by mucus alterations, including COPD. In the present study, the antioxidant activity of CLS was studied in vitro in three different oxygen radical producing systems, i.e. bronchoalveolar lavages (BAL) from patients affected by COPD, ultrasound treated human serum and cultured human lung endothelial cells challenged with elastase. BAL, exposed or not to different concentrations of CLS (1.5-30 mM), was assayed for free radical content by fluorometric analysis of DNA unwinding (FADU) or by cytochrome c reduction kinetics. Human serum was treated with ultrasound in the presence or absence of CLS (1.5, 2.5 mM) or N-acetyl cysteine (NAC; 4, 5 mM) and assayed for free radical content by FADU. Human endothelial cells cultured in vitro from pulmonary artery were incubated with elastase (0.3 IU/mL), in the presence or absence of glutathione (GSH; 0.65 mM) or CLS (0.16 mM). The supernatant was tested for cytochrome c reduction kinetics whereas cell homogenates were assessed for xanthine oxidase (XO) content by SDS-PAGE. Results showed that CLS is more effective as an in vitro scavenger in comparison to GSH and NAC. CLS reduced the damage of DNA from healthy donors exposed to COPD-BAL and was able to quench clastogenic activity induced in human serum by exposure to ultrasound at concentrations as low as 2.5 mM. NAC protect DNA from radical damage, starting from 5 mM. In human lung endothelial cells cultured in presence of elastase, CLS (0.16 mM) decreased xanthine oxidase activity. These results suggest that CLS could act by interfering with the conversion of xanthine dehydrogenase into superoxide-producing xanthine oxidase. The antioxidant activity of CLS could contribute to its therapeutic activity by reducing radical

  1. Seed-Specific Expression of a Lysine-Rich Protein Gene, GhLRP, from Cotton Significantly Increases the Lysine Content in Maize Seeds

    Directory of Open Access Journals (Sweden)

    Jing Yue

    2014-03-01

    Full Text Available Maize seed storage proteins are a major source of human and livestock consumption. However, these proteins have poor nutritional value, because they are deficient in lysine and tryptophan. Much research has been done to elevate the lysine content by reducing zein content or regulating the activities of key enzymes in lysine metabolism. Using the naturally lysine-rich protein genes, sb401 and SBgLR, from potato, we previously increased the lysine and protein contents of maize seeds. Here, we examined another natural lysine-rich protein gene, GhLRP, from cotton, which increased the lysine content of transgenic maize seeds at levels varying from 16.2% to 65.0% relative to the wild-type. The total protein content was not distinctly different, except in the six transgenic lines. The lipid and starch levels did not differ substantially in Gossypium hirsutum L. lysine-rich protein (GhLRP transgenic kernels when compared to wild-type. The agronomic characteristics of all the transgenic maize were also normal. GhLRP is a high-lysine protein candidate gene for increasing the lysine content of maize. This study provided a valuable model system for improving maize lysine content.

  2. Seed-specific expression of a lysine-rich protein gene, GhLRP, from cotton significantly increases the lysine content in maize seeds.

    Science.gov (United States)

    Yue, Jing; Li, Cong; Zhao, Qian; Zhu, Dengyun; Yu, Jingjuan

    2014-03-27

    Maize seed storage proteins are a major source of human and livestock consumption. However, these proteins have poor nutritional value, because they are deficient in lysine and tryptophan. Much research has been done to elevate the lysine content by reducing zein content or regulating the activities of key enzymes in lysine metabolism. Using the naturally lysine-rich protein genes, sb401 and SBgLR, from potato, we previously increased the lysine and protein contents of maize seeds. Here, we examined another natural lysine-rich protein gene, GhLRP, from cotton, which increased the lysine content of transgenic maize seeds at levels varying from 16.2% to 65.0% relative to the wild-type. The total protein content was not distinctly different, except in the six transgenic lines. The lipid and starch levels did not differ substantially in Gossypium hirsutum L. lysine-rich protein (GhLRP) transgenic kernels when compared to wild-type. The agronomic characteristics of all the transgenic maize were also normal. GhLRP is a high-lysine protein candidate gene for increasing the lysine content of maize. This study provided a valuable model system for improving maize lysine content.

  3. How modification of accessible lysines to phenylalanine modulates the structural and functional properties of horseradish peroxidase: a simulation study.

    Directory of Open Access Journals (Sweden)

    Leila Navapour

    Full Text Available Horseradish Peroxidase (HRP is one of the most studied peroxidases and a great number of chemical modifications and genetic manipulations have been carried out on its surface accessible residues to improve its stability and catalytic efficiency necessary for biotechnological applications. Most of the stabilized derivatives of HRP reported to date have involved chemical or genetic modifications of three surface-exposed lysines (K174, K232 and K241. In this computational study, we altered these lysines to phenylalanine residues to model those chemical modifications or genetic manipulations in which these positively charged lysines are converted to aromatic hydrophobic residues. Simulation results implied that upon these substitutions, the protein structure becomes less flexible. Stability gains are likely to be achieved due to the increased number of stable hydrogen bonds, improved heme-protein interactions and more integrated proximal Ca2+ binding pocket. We also found a new persistent hydrogen bond between the protein moiety (F174 and the heme prosthetic group as well as two stitching hydrogen bonds between the connecting loops GH and F'F″ in mutated HRP. However, detailed analysis of functionally related structural properties and dynamical features suggests reduced reactivity of the enzyme toward its substrates. Molecular dynamics simulations showed that substitutions narrow the bottle neck entry of peroxide substrate access channel and reduce the surface accessibility of the distal histidine (H42 and heme prosthetic group to the peroxide and aromatic substrates, respectively. Results also demonstrated that the area and volume of the aromatic-substrate binding pocket are significantly decreased upon modifications. Moreover, the hydrophobic patch functioning as a binding site or trap for reducing aromatic substrates is shrunk in mutated enzyme. Together, the results of this simulation study could provide possible structural clues to explain

  4. How modification of accessible lysines to phenylalanine modulates the structural and functional properties of horseradish peroxidase: a simulation study.

    Science.gov (United States)

    Navapour, Leila; Mogharrab, Navid; Amininasab, Mehriar

    2014-01-01

    Horseradish Peroxidase (HRP) is one of the most studied peroxidases and a great number of chemical modifications and genetic manipulations have been carried out on its surface accessible residues to improve its stability and catalytic efficiency necessary for biotechnological applications. Most of the stabilized derivatives of HRP reported to date have involved chemical or genetic modifications of three surface-exposed lysines (K174, K232 and K241). In this computational study, we altered these lysines to phenylalanine residues to model those chemical modifications or genetic manipulations in which these positively charged lysines are converted to aromatic hydrophobic residues. Simulation results implied that upon these substitutions, the protein structure becomes less flexible. Stability gains are likely to be achieved due to the increased number of stable hydrogen bonds, improved heme-protein interactions and more integrated proximal Ca2+ binding pocket. We also found a new persistent hydrogen bond between the protein moiety (F174) and the heme prosthetic group as well as two stitching hydrogen bonds between the connecting loops GH and F'F″ in mutated HRP. However, detailed analysis of functionally related structural properties and dynamical features suggests reduced reactivity of the enzyme toward its substrates. Molecular dynamics simulations showed that substitutions narrow the bottle neck entry of peroxide substrate access channel and reduce the surface accessibility of the distal histidine (H42) and heme prosthetic group to the peroxide and aromatic substrates, respectively. Results also demonstrated that the area and volume of the aromatic-substrate binding pocket are significantly decreased upon modifications. Moreover, the hydrophobic patch functioning as a binding site or trap for reducing aromatic substrates is shrunk in mutated enzyme. Together, the results of this simulation study could provide possible structural clues to explain those experimental

  5. Amine oxidation mediated by lysine-specific demethylase 1: quantum mechanics/molecular mechanics insights into mechanism and role of lysine 661.

    Science.gov (United States)

    Karasulu, Bora; Patil, Mahendra; Thiel, Walter

    2013-09-11

    We report classical molecular dynamics (MD) simulations and combined quantum mechanics/molecular mechanics (QM/MM) calculations to elucidate the catalytic mechanism of the rate-determining amine oxidation step in the lysine-specific demethylase 1 (LSD1)-catalyzed demethylation of the histone tail lysine (H3K4), with flavin adenine dinucleotide (FAD) acting as cofactor. The oxidation of substrate lysine (sLys) involves the cleavage of an α-CH bond accompanied by the transfer of a hydride ion equivalent to FAD, leading to an imine intermediate. This hydride transfer pathway is shown to be clearly favored for sLys oxidation over other proposed mechanisms, including the radical (or single-electron transfer) route as well as carbanion and polar-nucleophilic mechanisms. MD simulations on six NVT ensembles (covering different protonation states of sLys and K661 as well as the K661M mutant) identify two possible orientations of the reacting sLys and FAD subunits (called "downward" and "upward"). Calculations at the QM(B3LYP-D/6-31G*)/CHARMM22 level provide molecular-level insights into the mechanism, helping to understand how LSD1 achieves the activation of the rather inert methyl-CH bond in a metal-free environment. Factors such as proper alignment of sLys (downward orientation), transition-state stabilization (due to the protein environment and favorable orbital interactions), and product stabilization via adduct formation are found to be crucial for facilitating the oxidative α-CH bond cleavage. The current study also sheds light on the role of important active-site residues (Y761, K661, and W695) and of the conserved water-bridge motif. The steric influence of Y761 helps to position the reaction partners properly, K661 is predicted to get deprotonated prior to substrate binding and to act as an active-site base that accepts a proton from sLys to enable the subsequent amine oxidation, and the water bridge that is stabilized by K661 and W695 mediates this proton

  6. Ab Initio Calculations of Deuterium Isotope Effects on Chemical Shifts of Salt-Bridged Lysines

    DEFF Research Database (Denmark)

    Ullah, Saif; Ishimoto, Takayoshi; Williamson, Mike P.;

    2011-01-01

    Deuterium isotope effects measure the change in chemical shift on substitution of a proton by deuterium. They have been calculated by direct treatment of the H/D nuclear quantum effect using a multicomponent ab initio molecular orbital method based on a non-Born−Oppenheimer approximation....... This method enables the determination of both the electronic and the protonic (deuteronic) wave functions simultaneously and can directly calculate the geometrical difference induced by H/D isotope effects. The calculations show that the one-bond deuterium isotope effects on 15N nuclear shielding, 1Δ15N......(D), in ammonium and amines decrease as a counterion or water molecule moves closer to the nitrogen. 1Δ15N(D) and 2Δ1H(D) of the NH3+ groups of lysine residues in the B1 domain of protein G have been calculated using truncated side chains and also determined experimentally by NMR. Comparisons show...

  7. The Generation of Dehydroalanine Residues in Protonated Polypeptides: Ion/Ion Reactions for Introducing Selective Cleavages

    Science.gov (United States)

    Peng, Zhou; Bu, Jiexun; McLuckey, Scott A.

    2017-05-01

    We examine a gas-phase approach for converting a subset of amino acid residues in polypeptide cations to dehydroalanine (Dha). Subsequent activation of the modified polypeptide ions gives rise to specific cleavage N-terminal to the Dha residue. This process allows for the incorporation of selective cleavages in the structural characterization of polypeptide ions. An ion/ion reaction within the mass spectrometer between a multiply protonated polypeptide and the sulfate radical anion introduces a radical site into the multiply protonated polypeptide reactant. Subsequent collisional activation of the polypeptide radical cation gives rise to radical side chain loss from one of several particular amino acid side chains (e.g., leucine, asparagine, lysine, glutamine, and glutamic acid) to yield a Dha residue. The Dha residues facilitate preferential backbone cleavages to produce signature c- and z-ions, demonstrated with cations derived from melittin, mechano growth factor (MGF), and ubiquitin. The efficiencies for radical side chain loss and for subsequent generation of specific c- and z-ions have been examined as functions of precursor ion charge state and activation conditions using cations of ubiquitin as a model for a small protein. It is noted that these efficiencies are not strongly dependent on ion trap collisional activation conditions but are sensitive to precursor ion charge state. Moderate to low charge states show the greatest overall yields for the specific Dha cleavages, whereas small molecule losses (e.g., water/ammonia) dominate at the lowest charge states and proton catalyzed amide bond cleavages that give rise to b- and y-ions tend to dominate at high charge states.

  8. The Generation of Dehydroalanine Residues in Protonated Polypeptides: Ion/Ion Reactions for Introducing Selective Cleavages

    Science.gov (United States)

    Peng, Zhou; Bu, Jiexun; McLuckey, Scott A.

    2017-09-01

    We examine a gas-phase approach for converting a subset of amino acid residues in polypeptide cations to dehydroalanine (Dha). Subsequent activation of the modified polypeptide ions gives rise to specific cleavage N-terminal to the Dha residue. This process allows for the incorporation of selective cleavages in the structural characterization of polypeptide ions. An ion/ion reaction within the mass spectrometer between a multiply protonated polypeptide and the sulfate radical anion introduces a radical site into the multiply protonated polypeptide reactant. Subsequent collisional activation of the polypeptide radical cation gives rise to radical side chain loss from one of several particular amino acid side chains (e.g., leucine, asparagine, lysine, glutamine, and glutamic acid) to yield a Dha residue. The Dha residues facilitate preferential backbone cleavages to produce signature c- and z-ions, demonstrated with cations derived from melittin, mechano growth factor (MGF), and ubiquitin. The efficiencies for radical side chain loss and for subsequent generation of specific c- and z-ions have been examined as functions of precursor ion charge state and activation conditions using cations of ubiquitin as a model for a small protein. It is noted that these efficiencies are not strongly dependent on ion trap collisional activation conditions but are sensitive to precursor ion charge state. Moderate to low charge states show the greatest overall yields for the specific Dha cleavages, whereas small molecule losses (e.g., water/ammonia) dominate at the lowest charge states and proton catalyzed amide bond cleavages that give rise to b- and y-ions tend to dominate at high charge states. [Figure not available: see fulltext.

  9. Lysine-iron agar in the detection of Arizona cultures.

    Science.gov (United States)

    EDWARDS, P R; FIFE, M A

    1961-11-01

    A lysine-iron agar is described and recommended for the detection of Arizona strains which ferment lactose rapidly. Black colonies which appear on bismuth sulfite agar should be transferred to the medium. Salmonellae and Arizona cultures produce a distinctive reaction since they are the only recognized groups of enteric bacteria which regularly produce lysine decarboxylase rapidly and form large amounts of hydrogen sulfide. Use of the medium is particularly recommended in the examination of specimens from enteric infections in which shigellae and salmonellae are not detected.

  10. Sugar Substrates for l-Lysine Fermentation by Ustilago maydis

    Science.gov (United States)

    Sánchez-Marroquín, A.; Ledezma, M.; Carreño, R.

    1970-01-01

    The extracellular production of l-lysine in media with cane sugar, blackstrap molasses, or clarified sugar-cane juice by a previously obtained mutant of Ustilago maydis was studied. Enzymatically inverted clarified juice (medium J-3) gave 2.9 g of lysine per liter under the following conditions: inoculum, 5%; pH 5.8; temperature, 30 C; KLa in the fermentors, 0.41 mmoles of O2 per liter per min; fermentation time, 72 hr. The concentrate, obtained by direct evaporation and drying of the fermentation broth, could be used as a possible feed supplement because of its amino-acid and vitamin content. PMID:5485081

  11. pH-Sensitive surfactants from lysine: assessment of their cytotoxicity and environmental behavior.

    Science.gov (United States)

    Colomer, Aurora; Pinazo, Aurora; García, Maria Teresa; Mitjans, Montserrat; Vinardell, M Pilar; Infante, Maria Rosa; Martínez, Verónica; Pérez, Lourdes

    2012-04-10

    The toxicity and environmental behavior of new pH-sensitive surfactants from lysine are presented. Three different chemical structures are studied: surfactants with one amino acid and one alkyl chain, surfactants with two amino acids on the polar head and one alkyl chain, and gemini surfactants. The pH sensitivity of these compounds can be tuned by modifying their chemical structures. Cytotoxicity has been evaluated using erythrocytes and fibroblast cells. The toxic effects against these cells depend on the hydrophobicity of the molecules as well as their cationic charge density. The effect of hydrophobicity and cationic charge density on toxicity is different for each type of cells. For erythrocytes, the toxicity increases as hydrophobicity and charge density increases. Nevertheless, for fibroblasts cationic charge density affects cytotoxicity in the opposite way: the higher charge density, the lower the toxicity. The effect of the pH on hemolysis has been evaluated in detail. The aquatic toxicity was established using Daphnia magna . All surfactants yielded EC(50) values considerably higher than that reported for cationic surfactants based on quaternary ammonium groups. Finally, their biodegradability was evaluated using the CO(2) headspace test (ISO 14593). These lysine derivatives showed high levels of biodegradation under aerobic conditions and can be classified as "readily biodegradable compounds".

  12. Low plasma carnitine in patients on prolonged total parenteral nutrition: association with low plasma lysine.

    Science.gov (United States)

    Berner, Y N; Larchian, W A; Lowry, S F; Nicroa, R R; Brennan, M F; Shike, M

    1990-01-01

    Plasma carnitine levels were determined in 17 patients maintained on long-term total parenteral nutrition (TPN) for a mean (+/- SEM) period of 69 +/- 11 months (range 12-196). All had severe malabsorption and were dependent on intravenous feeding. Plasma carnitine was determined by a modified Cederblad enzymatic method. Mean plasma carnitine was significantly below the mean normal for females (p less than 0.02) and borderline low for males (p = 0.07). In six patients the levels were below the low normal range, and in five others they were at the lowest levels of normal. Of the six patients with normal levels, three had elevated serum creatinine, indicating renal dysfunction which may by itself elevate plasma carnitine. In 10 patients the plasma levels of lysine (a carnitine precursor) were determined and found to be lower than normal (p less than 0.05). Plasma carnitine levels correlated positively with serum albumin (r = 0.62, p less than 0.05), and negatively with serum alkaline phosphatase (r = -0.64, p less than 0.05). Thus, patients maintained on long-term TPN may have low plasma carnitine, which could represent carnitine deficiency. The low plasma carnitine may be related to a deficiency of the carnitine precursor lysine. Further studies are required to determine the significance of the low plasma carnitine and whether carnitine supplementation should be required in long-term TPN.

  13. Effect of Selected Plant Extracts and D- and L-Lysine on the Cyanobacterium Microcystis aeruginosa

    National Research Council Canada - National Science Library

    Lurling, M; Van Oosterhout, F

    2014-01-01

    We tested extracts from Fructus mume, Salvia miltiorrhiza and Moringa oleifera as well as L-lysine and D-Lysine as curative measures to rapidly suppress the cyanobacterium Microcystis aeruginosa NIVA-CYA 43...

  14. Evidence supporting a role for Nε-(3-formyl-3,4-dehydropiperidino)lysine accumulation in Müller glia dysfunction and death in diabetic retinopathy

    Science.gov (United States)

    Yong, Phaik Har; Zong, Hongliang; Medina, Reinhold J.; Limb, G. Astrid; Uchida, Koji; Stitt, Alan W.

    2010-01-01

    Purpose Recent evidence suggests that neuroglial dysfunction and degeneration contributes to the etiology and progression of diabetic retinopathy. Advanced lipoxidation end products (ALEs) have been implicated in the pathology of various diseases, including diabetes and several neurodegenerative disorders. The purpose of the present study was to investigate the possible link between the accumulation of ALEs and neuroretinal changes in diabetic retinopathy. Methods Retinal sections obtained from diabetic rats and age-matched controls were processed for immunohistochemistry using antibodies against several well defined ALEs. In vitro experiments were also performed using a human Müller (Moorfields/Institute of Ophthalmology-Müller 1 [MIO-M1]) glia cell line. Western blot analysis was used to measure the accumulation of the acrolein-derived ALE adduct Nε-(3-formyl-3,4-dehydropiperidino)lysine (FDP-lysine) in Müller cells preincubated with FDP-lysine-modified human serum albumin (FDP-lysine-HSA). Responses of Müller cells to FDP-lysine accumulation were investigated by analyzing changes in the protein expression of heme oxygenase-1 (HO-1), glial fibrillary acidic protein (GFAP), and the inwardly rectifying potassium channel Kir4.1. In addition, mRNA expression levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), and tumor necrosis factor-α (TNFα) were determined by reverse transcriptase PCR (RT–PCR). Apoptotic cell death was evaluated by fluorescence-activated cell sorting (FACS) analysis after staining with fluorescein isothiocyanate (FITC)-labeled annexin V and propidium iodide. Results No significant differences in the levels of malondialdehyde-, 4-hydroxy-2-nonenal-, and 4-hydroxyhexenal-derived ALEs were evident between control and diabetic retinas after 4 months of diabetes. By contrast, FDP-lysine immunoreactivity was markedly increased in the Müller glia of diabetic rats. Time-course studies revealed that FDP-lysine initially

  15. SITE-SPECIFIC LABELING OF A PROTEIN LYSINE RESIDUE BY NOVEL KINETIC LABELING COMBINATORIAL LIBRARIES

    Directory of Open Access Journals (Sweden)

    Allen Krantz

    2014-03-01

    Full Text Available The first example of a kinetic labeling library designed to enable the discovery of affinity labels is presented. Each library component (1 consists of a variable peptidyl component linked to a biotinyl moiety by a 4-mercaptobenzoyl linker in thioester format. We demonstrate that an affinity label can be uncovered by measuring reaction rates between library pools and the protein target, human serum albumin (HSA and identifying significant outliers. By choosing peptide functionality compatible with a potentially reactive thioester labeling entity, libraries can be screened in pools. It is noteworthy that a limited subset of amino acids (R, S, E, F, Y, l, M, W, and Q that compose the affinity moiety is sufficient to produce rate variances that guide the discovery process. After two rounds of deconvolution, J-FLYEE-NH2 (7-E emerges as a bona fide affinity label of HSA. Unlike known affinity labels, the affinity moiety is not retained in the protein product, but is extruded upon acylation of the protein. This feature affords a method of introducing various payloads, without extraneous elements, onto protein frameworks.

  16. [Modification of the lysine-iron agar (author's transl)].

    Science.gov (United States)

    Wauters, G

    1975-12-01

    The addition of L-phenylalanine to the lysine-iron agar described by Edwards and Fife ]1] allows a more valuable screening of the Proteus group based on its deamination properties. Some minor modifications of the indicator and thiosulfate content lead to improve and earlier recording of the results.

  17. Detection of salt bridges to lysines in solution in barnase

    DEFF Research Database (Denmark)

    Hansen, Poul Erik; Williamson, Michael P.; Hounslow, Andrea M.

    2013-01-01

    We show that salt bridges involving lysines can be detected by deuterium isotope effects on NMR chemical shifts of the sidechain amine. Lys27 in the ribonuclease barnase is salt bridged, and mutation of Arg69 to Lys retains a partially buried salt bridge. The salt bridges are functionally important....

  18. Requirement of the laying hen for apparent fecal digestible lysine

    NARCIS (Netherlands)

    Schutte, J.B.; Smink, W.

    1998-01-01

    A study was conducted to determine the requirement for lysine of a White Leghorn strain of hens with a body weight of approximately 1,600 g. Before starting the experiment, apparent fecal digestibility of amino acids of the basal diet was determined in an in vivo digestibility trial with six individ

  19. Predicting post-translational lysine acetylation using support vector machines

    DEFF Research Database (Denmark)

    Gnad, Florian; Ren, Shubin; Choudhary, Chunaram

    2010-01-01

    spectrometry to identify 3600 lysine acetylation sites on 1750 human proteins covering most of the previously annotated sites and providing the most comprehensive acetylome so far. This dataset should provide an excellent source to train support vector machines (SVMs) allowing the high accuracy in silico...

  20. effects of dietary chromium tripicolinate and lysine on growth ...

    African Journals Online (AJOL)

    AISA

    Six traitements ont été répétés quatre fois, avec quatre porcs par répétition. Au cours de cette ... The potential capability of lysine to improve ... (chromium picolinate) on animal productivity has ... cholesterol (Sigma, 1989a), and total proteins.

  1. Amino acid nutrition beyond methionine and lysine for milk protein

    Science.gov (United States)

    Amino acids are involved in many important physiological processes affecting the production, health, and reproduction of high-producing dairy cows. Most research and recommendations for lactating dairy cows has focused on methionine and lysine for increasing milk protein yield. This is because these...

  2. File list: Oth.Gon.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Gon.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Gonad SRX1060...566,SRX1060567,SRX1060557 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Gon.50.Crotonyl_lysine.AllCell.bed ...

  3. File list: His.Bon.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Bo...ne http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bon.50.Pan_lysine_crotonylation.AllCell.bed ...

  4. File list: His.Bld.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Bl...ood http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bld.05.Pan_lysine_crotonylation.AllCell.bed ...

  5. File list: His.Utr.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Utr.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ut...erus http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Utr.05.Pan_lysine_crotonylation.AllCell.bed ...

  6. File list: His.Emb.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Emb.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Em...bryo http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Emb.50.Pan_lysine_crotonylation.AllCell.bed ...

  7. File list: His.PSC.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Pl...uripotent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.50.Pan_lysine_crotonylation.AllCell.bed ...

  8. File list: His.ALL.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Al...l cell types SRX099897,SRX099894 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.ALL.10.Pan_lysine_crotonylation.AllCell.bed ...

  9. File list: His.Epd.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation E...pidermis http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Epd.50.Pan_lysine_crotonylation.AllCell.bed ...

  10. File list: His.Plc.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Plc.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...lacenta http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Plc.50.Pan_lysine_crotonylation.AllCell.bed ...

  11. File list: His.Bon.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Bo...ne http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bon.20.Pan_lysine_crotonylation.AllCell.bed ...

  12. File list: His.Neu.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Neu.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation N...eural http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Neu.50.Pan_lysine_crotonylation.AllCell.bed ...

  13. File list: His.Bld.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Bl...ood http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bld.10.Pan_lysine_crotonylation.AllCell.bed ...

  14. File list: His.CDV.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.CDV.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ca...rdiovascular http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.CDV.50.Pan_lysine_crotonylation.AllCell.bed ...

  15. File list: His.Oth.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ot...hers http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Oth.10.Pan_lysine_crotonylation.AllCell.bed ...

  16. File list: His.Adp.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation A...dipocyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.Pan_lysine_crotonylation.AllCell.bed ...

  17. File list: His.Dig.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation D...igestive tract http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Dig.05.Pan_lysine_crotonylation.AllCell.bed ...

  18. File list: His.Gon.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Gon.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation G...onad http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Gon.05.Pan_lysine_crotonylation.AllCell.bed ...

  19. File list: His.Dig.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Di...gestive tract http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Dig.20.Pan_lysine_crotonylation.AllCell.bed ...

  20. File list: Oth.Dig.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Dig.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Digestive tra...ct http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Dig.50.Crotonyl_lysine.AllCell.bed ...

  1. File list: His.Liv.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Li...ver http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.10.Pan_lysine_crotonylation.AllCell.bed ...

  2. File list: His.Kid.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Kid.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation K...idney http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Kid.20.Pan_lysine_crotonylation.AllCell.bed ...

  3. File list: His.Pan.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...ancreas http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Pan.50.Pan_lysine_crotonylation.AllCell.bed ...

  4. File list: His.Unc.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Un...classified http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Unc.50.Pan_lysine_crotonylation.AllCell.bed ...

  5. File list: His.Kid.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Kid.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ki...dney http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Kid.50.Pan_lysine_crotonylation.AllCell.bed ...

  6. File list: His.Lng.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation L...ung SRX099891 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Lng.05.Pan_lysine_crotonylation.AllCell.bed ...

  7. File list: His.Liv.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation L...iver http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Liv.20.Pan_lysine_crotonylation.AllCell.bed ...

  8. File list: His.PSC.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...luripotent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.50.Pan_lysine_crotonylation.AllCell.bed ...

  9. File list: His.CDV.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.CDV.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ca...rdiovascular http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.CDV.05.Pan_lysine_crotonylation.AllCell.bed ...

  10. File list: His.Lng.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Lu...ng http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Lng.20.Pan_lysine_crotonylation.AllCell.bed ...

  11. File list: His.ALL.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation A...ll cell types SRX099891 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.ALL.10.Pan_lysine_crotonylation.AllCell.bed ...

  12. File list: His.Neu.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Neu.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ne...ural http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Neu.20.Pan_lysine_crotonylation.AllCell.bed ...

  13. File list: His.Bld.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Bl...ood http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bld.20.Pan_lysine_crotonylation.AllCell.bed ...

  14. File list: His.Liv.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Li...ver http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.20.Pan_lysine_crotonylation.AllCell.bed ...

  15. File list: His.Utr.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Utr.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ut...erus http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Utr.20.Pan_lysine_crotonylation.AllCell.bed ...

  16. File list: Oth.Gon.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Gon.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Gonad SRX1060...566,SRX1060567,SRX1060557 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Gon.20.Crotonyl_lysine.AllCell.bed ...

  17. File list: His.Myo.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation M...uscle http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Myo.50.Pan_lysine_crotonylation.AllCell.bed ...

  18. File list: His.Plc.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Plc.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...lacenta http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Plc.05.Pan_lysine_crotonylation.AllCell.bed ...

  19. File list: His.Unc.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation U...nclassified http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Unc.20.Pan_lysine_crotonylation.AllCell.bed ...

  20. File list: His.Brs.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Brs.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Br...east http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Brs.05.Pan_lysine_crotonylation.AllCell.bed ...

  1. File list: Oth.NoD.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.NoD.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine No descriptio...n http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.NoD.50.Crotonyl_lysine.AllCell.bed ...

  2. File list: His.Pan.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Pa...ncreas http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Pan.20.Pan_lysine_crotonylation.AllCell.bed ...

  3. File list: His.Liv.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation L...iver http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Liv.05.Pan_lysine_crotonylation.AllCell.bed ...

  4. File list: His.Brs.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Brs.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...reast http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Brs.20.Pan_lysine_crotonylation.AllCell.bed ...

  5. File list: His.Gon.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Gon.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation G...onad http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Gon.20.Pan_lysine_crotonylation.AllCell.bed ...

  6. File list: Oth.Adp.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Adipocyte htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.20.Crotonyl_lysine.AllCell.bed ...

  7. File list: His.ALL.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Al...l cell types SRX099894,SRX099897 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.ALL.20.Pan_lysine_crotonylation.AllCell.bed ...

  8. File list: His.Bon.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...one http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bon.10.Pan_lysine_crotonylation.AllCell.bed ...

  9. File list: His.Myo.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Mu...scle http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Myo.50.Pan_lysine_crotonylation.AllCell.bed ...

  10. File list: His.Bld.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...lood http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.10.Pan_lysine_crotonylation.AllCell.bed ...

  11. File list: His.Liv.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Li...ver http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.05.Pan_lysine_crotonylation.AllCell.bed ...

  12. File list: His.Epd.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ep...idermis http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Epd.20.Pan_lysine_crotonylation.AllCell.bed ...

  13. File list: His.Kid.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Kid.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation K...idney http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Kid.50.Pan_lysine_crotonylation.AllCell.bed ...

  14. File list: His.Lng.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Lu...ng http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Lng.50.Pan_lysine_crotonylation.AllCell.bed ...

  15. File list: His.Lng.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation L...ung SRX099891 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Lng.10.Pan_lysine_crotonylation.AllCell.bed ...

  16. File list: His.PSC.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...luripotent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.10.Pan_lysine_crotonylation.AllCell.bed ...

  17. File list: His.Emb.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Emb.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Em...bryo http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Emb.10.Pan_lysine_crotonylation.AllCell.bed ...

  18. File list: His.Kid.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Kid.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation K...idney http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Kid.05.Pan_lysine_crotonylation.AllCell.bed ...

  19. File list: His.Unc.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Un...classified http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Unc.20.Pan_lysine_crotonylation.AllCell.bed ...

  20. File list: His.Unc.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation U...nclassified http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Unc.50.Pan_lysine_crotonylation.AllCell.bed ...

  1. File list: His.Kid.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Kid.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ki...dney http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Kid.05.Pan_lysine_crotonylation.AllCell.bed ...

  2. File list: His.Pan.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...ancreas http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Pan.05.Pan_lysine_crotonylation.AllCell.bed ...

  3. File list: His.Adp.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation A...dipocyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.10.Pan_lysine_crotonylation.AllCell.bed ...

  4. File list: His.Prs.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...rostate http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Prs.05.Pan_lysine_crotonylation.AllCell.bed ...

  5. File list: His.Utr.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Utr.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation U...terus http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Utr.50.Pan_lysine_crotonylation.AllCell.bed ...

  6. File list: His.Bld.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...lood http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.05.Pan_lysine_crotonylation.AllCell.bed ...

  7. File list: Oth.Dig.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Dig.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Digestive tra...ct http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Dig.05.Crotonyl_lysine.AllCell.bed ...

  8. File list: His.Kid.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Kid.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation K...idney http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Kid.10.Pan_lysine_crotonylation.AllCell.bed ...

  9. File list: Oth.NoD.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.NoD.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine No descriptio...n http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.NoD.20.Crotonyl_lysine.AllCell.bed ...

  10. File list: Oth.EmF.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.EmF.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Embryonic fib...roblast http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.EmF.05.Crotonyl_lysine.AllCell.bed ...

  11. File list: His.CDV.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.CDV.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ca...rdiovascular http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.CDV.20.Pan_lysine_crotonylation.AllCell.bed ...

  12. File list: His.Pan.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Pa...ncreas http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Pan.10.Pan_lysine_crotonylation.AllCell.bed ...

  13. File list: His.PSC.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Pl...uripotent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.20.Pan_lysine_crotonylation.AllCell.bed ...

  14. File list: His.Dig.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Di...gestive tract http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Dig.10.Pan_lysine_crotonylation.AllCell.bed ...

  15. File list: His.Brs.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Brs.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...reast http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Brs.10.Pan_lysine_crotonylation.AllCell.bed ...

  16. File list: His.Bld.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...lood http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.50.Pan_lysine_crotonylation.AllCell.bed ...

  17. File list: Oth.Dig.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Dig.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Digestive tra...ct http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Dig.20.Crotonyl_lysine.AllCell.bed ...

  18. File list: His.Myo.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation M...uscle http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Myo.05.Pan_lysine_crotonylation.AllCell.bed ...

  19. File list: His.Adp.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ad...ipocyte http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.50.Pan_lysine_crotonylation.AllCell.bed ...

  20. File list: His.Emb.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Emb.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Em...bryo http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Emb.20.Pan_lysine_crotonylation.AllCell.bed ...

  1. File list: His.Oth.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation O...thers http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Oth.20.Pan_lysine_crotonylation.AllCell.bed ...

  2. File list: His.Plc.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Plc.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...lacenta http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Plc.20.Pan_lysine_crotonylation.AllCell.bed ...

  3. File list: His.Myo.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation M...uscle http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Myo.10.Pan_lysine_crotonylation.AllCell.bed ...

  4. File list: Oth.CDV.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.CDV.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Cardiovascula...r http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.CDV.20.Crotonyl_lysine.AllCell.bed ...

  5. File list: His.Bon.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...one http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bon.05.Pan_lysine_crotonylation.AllCell.bed ...

  6. File list: His.Dig.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Diges...tive tract http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Dig.50.Pan_lysine_acetylation.AllCell.bed ...

  7. File list: His.Pan.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Pancre...as http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Pan.05.Pan_lysine_acetylation.AllCell.bed ...

  8. File list: His.Unc.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Uncla...ssified http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Unc.10.Pan_lysine_acetylation.AllCell.bed ...

  9. File list: His.Pan.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Pancr...eas http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Pan.20.Pan_lysine_acetylation.AllCell.bed ...

  10. File list: His.Neu.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Neu.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Neural... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Neu.20.Pan_lysine_acetylation.AllCell.bed ...

  11. File list: His.Epd.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.05.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Epide...rmis http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Epd.05.Pan_lysine_acetylation.AllCell.bed ...

  12. File list: His.Brs.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Brs.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Breast... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Brs.20.Pan_lysine_acetylation.AllCell.bed ...

  13. File list: His.Dig.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.05.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Diges...tive tract http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Dig.05.Pan_lysine_acetylation.AllCell.bed ...

  14. File list: His.Prs.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Prosta...te http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Prs.20.Pan_lysine_acetylation.AllCell.bed ...

  15. File list: His.Liv.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Liver ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.50.Pan_lysine_acetylation.AllCell.bed ...

  16. File list: His.Myo.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Muscle... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Myo.20.Pan_lysine_acetylation.AllCell.bed ...

  17. File list: His.ALL.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation All ce...ll types SRX099893,SRX099896 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.ALL.10.Pan_lysine_acetylation.AllCell.bed ...

  18. File list: His.Emb.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Emb.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Embryo... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Emb.10.Pan_lysine_acetylation.AllCell.bed ...

  19. File list: His.Pan.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Pancr...eas http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Pan.10.Pan_lysine_acetylation.AllCell.bed ...

  20. File list: His.Unc.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Unclas...sified http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Unc.05.Pan_lysine_acetylation.AllCell.bed ...

  1. File list: His.Adp.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Adipo...cyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.Pan_lysine_acetylation.AllCell.bed ...

  2. File list: His.Neu.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Neu.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Neura...l http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Neu.10.Pan_lysine_acetylation.AllCell.bed ...

  3. File list: His.Utr.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Utr.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Uterus... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Utr.20.Pan_lysine_acetylation.AllCell.bed ...

  4. File list: His.ALL.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation All ce...ll types SRX099893,SRX099896 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.ALL.50.Pan_lysine_acetylation.AllCell.bed ...

  5. File list: His.Bld.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Blood... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.10.Pan_lysine_acetylation.AllCell.bed ...

  6. File list: His.CDV.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.CDV.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Cardi...ovascular http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.CDV.20.Pan_lysine_acetylation.AllCell.bed ...

  7. File list: His.Bld.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Blood ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bld.20.Pan_lysine_acetylation.AllCell.bed ...

  8. File list: His.Bld.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Blood ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bld.50.Pan_lysine_acetylation.AllCell.bed ...

  9. File list: His.Dig.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Diges...tive tract http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Dig.20.Pan_lysine_acetylation.AllCell.bed ...

  10. File list: His.Oth.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Others... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Oth.10.Pan_lysine_acetylation.AllCell.bed ...

  11. File list: His.Prs.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Prost...ate http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Prs.20.Pan_lysine_acetylation.AllCell.bed ...

  12. File list: His.Epd.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Epider...mis http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Epd.20.Pan_lysine_acetylation.AllCell.bed ...

  13. File list: His.Neu.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Neu.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Neural... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Neu.10.Pan_lysine_acetylation.AllCell.bed ...

  14. File list: His.Unc.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.05.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Uncla...ssified http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Unc.05.Pan_lysine_acetylation.AllCell.bed ...

  15. File list: His.Brs.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Brs.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Breas...t http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Brs.20.Pan_lysine_acetylation.AllCell.bed ...

  16. File list: His.Lng.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Lung h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Lng.20.Pan_lysine_acetylation.AllCell.bed ...

  17. File list: His.Kid.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Kid.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Kidney... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Kid.05.Pan_lysine_acetylation.AllCell.bed ...

  18. File list: His.Adp.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Adipo...cyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.20.Pan_lysine_acetylation.AllCell.bed ...

  19. File list: His.Oth.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Others... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Oth.20.Pan_lysine_acetylation.AllCell.bed ...

  20. File list: His.Bon.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Bone h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bon.05.Pan_lysine_acetylation.AllCell.bed ...

  1. File list: His.Bld.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Blood ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bld.10.Pan_lysine_acetylation.AllCell.bed ...

  2. File list: His.Adp.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Adipo...cyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.10.Pan_lysine_acetylation.AllCell.bed ...

  3. File list: His.Adp.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Adipoc...yte http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.10.Pan_lysine_acetylation.AllCell.bed ...

  4. File list: His.CDV.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.CDV.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Cardio...vascular http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.CDV.50.Pan_lysine_acetylation.AllCell.bed ...

  5. File list: His.Neu.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Neu.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Neural... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Neu.50.Pan_lysine_acetylation.AllCell.bed ...

  6. File list: His.Lng.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Lung h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Lng.50.Pan_lysine_acetylation.AllCell.bed ...

  7. File list: His.PSC.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Plurip...otent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.10.Pan_lysine_acetylation.AllCell.bed ...

  8. File list: His.Plc.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Plc.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Placen...ta http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Plc.20.Pan_lysine_acetylation.AllCell.bed ...

  9. File list: His.Oth.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Others... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Oth.50.Pan_lysine_acetylation.AllCell.bed ...

  10. File list: His.Oth.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Others... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Oth.05.Pan_lysine_acetylation.AllCell.bed ...

  11. File list: His.Lng.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Lung h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Lng.05.Pan_lysine_acetylation.AllCell.bed ...

  12. File list: His.Prs.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Prost...ate http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Prs.50.Pan_lysine_acetylation.AllCell.bed ...

  13. File list: His.Myo.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Muscl...e http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Myo.20.Pan_lysine_acetylation.AllCell.bed ...

  14. File list: His.Unc.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Unclas...sified http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Unc.50.Pan_lysine_acetylation.AllCell.bed ...

  15. File list: His.Epd.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Epide...rmis http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Epd.20.Pan_lysine_acetylation.AllCell.bed ...

  16. File list: His.Prs.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Prosta...te http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Prs.05.Pan_lysine_acetylation.AllCell.bed ...

  17. File list: His.Bld.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.05.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Blood... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.05.Pan_lysine_acetylation.AllCell.bed ...

  18. File list: His.ALL.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation All ce...ll types SRX099893,SRX099896 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.ALL.05.Pan_lysine_acetylation.AllCell.bed ...

  19. File list: His.Neu.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Neu.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Neura...l http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Neu.20.Pan_lysine_acetylation.AllCell.bed ...

  20. File list: His.PSC.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Plurip...otent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.20.Pan_lysine_acetylation.AllCell.bed ...

  1. File list: His.Pan.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.05.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Pancr...eas http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Pan.05.Pan_lysine_acetylation.AllCell.bed ...

  2. File list: His.ALL.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation All c...ell types SRX099890 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.ALL.20.Pan_lysine_acetylation.AllCell.bed ...

  3. File list: His.Liv.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Liver ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.20.Pan_lysine_acetylation.AllCell.bed ...

  4. File list: His.Gon.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Gon.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Gonad... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Gon.50.Pan_lysine_acetylation.AllCell.bed ...

  5. File list: His.ALL.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation All c...ell types SRX099890 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.ALL.10.Pan_lysine_acetylation.AllCell.bed ...

  6. File list: His.PSC.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Pluri...potent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.50.Pan_lysine_acetylation.AllCell.bed ...

  7. File list: His.Prs.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Prosta...te http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Prs.50.Pan_lysine_acetylation.AllCell.bed ...

  8. File list: His.CDV.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.CDV.05.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Cardi...ovascular http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.CDV.05.Pan_lysine_acetylation.AllCell.bed ...

  9. File list: His.Gon.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Gon.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Gonad ...SRX099893,SRX099896 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Gon.10.Pan_lysine_acetylation.AllCell.bed ...

  10. File list: His.Bon.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Bone ...http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bon.20.Pan_lysine_acetylation.AllCell.bed ...

  11. File list: His.Dig.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Digest...ive tract http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Dig.10.Pan_lysine_acetylation.AllCell.bed ...

  12. File list: His.Bld.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Blood... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.20.Pan_lysine_acetylation.AllCell.bed ...

  13. File list: His.Unc.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Uncla...ssified http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Unc.50.Pan_lysine_acetylation.AllCell.bed ...

  14. File list: His.Utr.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Utr.05.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Uteru...s http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Utr.05.Pan_lysine_acetylation.AllCell.bed ...

  15. File list: His.Liv.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Liver ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.05.Pan_lysine_acetylation.AllCell.bed ...

  16. File list: His.Lng.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Lung h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Lng.10.Pan_lysine_acetylation.AllCell.bed ...

  17. File list: His.Liv.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Liver ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.10.Pan_lysine_acetylation.AllCell.bed ...

  18. File list: His.Bld.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Blood ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bld.05.Pan_lysine_acetylation.AllCell.bed ...

  19. File list: His.Epd.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Epider...mis http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Epd.10.Pan_lysine_acetylation.AllCell.bed ...

  20. File list: His.Pan.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Pancr...eas http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Pan.50.Pan_lysine_acetylation.AllCell.bed ...

  1. File list: His.PSC.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Plurip...otent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.PSC.50.Pan_lysine_acetylation.AllCell.bed ...

  2. File list: His.Plc.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Plc.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Placen...ta http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Plc.10.Pan_lysine_acetylation.AllCell.bed ...

  3. File list: His.Plc.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Plc.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Place...nta http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Plc.10.Pan_lysine_acetylation.AllCell.bed ...

  4. File list: His.Prs.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Prost...ate http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Prs.10.Pan_lysine_acetylation.AllCell.bed ...

  5. File list: His.Bon.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Bone h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bon.50.Pan_lysine_acetylation.AllCell.bed ...

  6. File list: His.Lng.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Lung ...SRX099890 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Lng.20.Pan_lysine_acetylation.AllCell.bed ...

  7. File list: His.Unc.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Unclas...sified http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Unc.10.Pan_lysine_acetylation.AllCell.bed ...

  8. File list: His.Utr.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Utr.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Uteru...s http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Utr.10.Pan_lysine_acetylation.AllCell.bed ...

  9. File list: His.Adp.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Adipoc...yte http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.20.Pan_lysine_acetylation.AllCell.bed ...

  10. File list: His.Gon.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Gon.50.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Gonad ...SRX099893,SRX099896 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Gon.50.Pan_lysine_acetylation.AllCell.bed ...

  11. File list: His.Dig.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Digest...ive tract http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Dig.05.Pan_lysine_acetylation.AllCell.bed ...

  12. File list: His.Dig.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Dig.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Diges...tive tract http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Dig.10.Pan_lysine_acetylation.AllCell.bed ...

  13. File list: His.Brs.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Brs.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Breast... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Brs.10.Pan_lysine_acetylation.AllCell.bed ...

  14. File list: His.Myo.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Muscle... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Myo.10.Pan_lysine_acetylation.AllCell.bed ...

  15. File list: His.CDV.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.CDV.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Cardio...vascular http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.CDV.10.Pan_lysine_acetylation.AllCell.bed ...

  16. File list: His.Bon.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Bone h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bon.20.Pan_lysine_acetylation.AllCell.bed ...

  17. File list: His.Epd.05.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.05.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Epider...mis http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Epd.05.Pan_lysine_acetylation.AllCell.bed ...

  18. File list: His.Oth.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Other...s http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Oth.10.Pan_lysine_acetylation.AllCell.bed ...

  19. File list: Oth.Adp.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Adipocyte htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.50.Crotonyl_lysine.AllCell.bed ...

  20. File list: Oth.NoD.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.NoD.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine No descriptio...n http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.NoD.05.Crotonyl_lysine.AllCell.bed ...

  1. File list: Oth.PSC.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Pluripotent s...tem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.50.Crotonyl_lysine.AllCell.bed ...

  2. File list: Oth.PSC.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Pluripotent s...tem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.10.Crotonyl_lysine.AllCell.bed ...

  3. File list: His.Myo.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Myo.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Mu...scle http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Myo.05.Pan_lysine_crotonylation.AllCell.bed ...

  4. File list: His.Prs.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Prs.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Pr...ostate http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Prs.50.Pan_lysine_crotonylation.AllCell.bed ...

  5. File list: His.ALL.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.ALL.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation A...ll cell types SRX099891 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.ALL.20.Pan_lysine_crotonylation.AllCell.bed ...

  6. File list: His.Gon.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Gon.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Go...nad SRX099894,SRX099897 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Gon.20.Pan_lysine_crotonylation.AllCell.bed ...

  7. File list: His.Oth.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.10.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation O...thers http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Oth.10.Pan_lysine_crotonylation.AllCell.bed ...

  8. File list: Oth.EmF.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.EmF.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Embryonic fib...roblast http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.EmF.20.Crotonyl_lysine.AllCell.bed ...

  9. File list: His.PSC.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.PSC.05.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation P...luripotent stem cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.05.Pan_lysine_crotonylation.AllCell.bed ...

  10. File list: His.Lng.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Lng.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Lu...ng http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Lng.10.Pan_lysine_crotonylation.AllCell.bed ...

  11. File list: His.Emb.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Emb.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Em...bryo http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Emb.05.Pan_lysine_crotonylation.AllCell.bed ...

  12. File list: Oth.CDV.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.CDV.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Cardiovascula...r http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.CDV.05.Crotonyl_lysine.AllCell.bed ...

  13. File list: His.Epd.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Ep...idermis http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Epd.10.Pan_lysine_crotonylation.AllCell.bed ...

  14. File list: His.Bon.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Bo...ne http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Bon.10.Pan_lysine_crotonylation.AllCell.bed ...

  15. File list: His.Bld.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bld.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...lood http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bld.20.Pan_lysine_crotonylation.AllCell.bed ...

  16. File list: His.Spl.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Spl.20.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Sp...leen http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Spl.20.Pan_lysine_crotonylation.AllCell.bed ...

  17. File list: His.Liv.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Liv.50.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Li...ver http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Liv.50.Pan_lysine_crotonylation.AllCell.bed ...

  18. File list: His.Pan.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Pan.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Pa...ncreas http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Pan.05.Pan_lysine_crotonylation.AllCell.bed ...

  19. File list: His.Unc.05.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Unc.05.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Un...classified http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Unc.05.Pan_lysine_crotonylation.AllCell.bed ...

  20. Optimization of lysine production in Corynebacteriumglutamicum ATCC15032 by Response surface methodology

    Directory of Open Access Journals (Sweden)

    Mehrnaz Haghi

    2017-03-01

    Discussion and conclusion: According to the results, the proposed culture media by response surface methodology causes 1400 times increase in the lysine production compared with M9 culture media and methionine had an important role in the production of lysine, probably by inhibiting the other metabolic pathway which has common metabolic precursor with lysine production metabolic pathway.

  1. File list: Oth.Gon.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Gon.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Gonad SRX1060...567,SRX1060566,SRX1060557 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Gon.10.Crotonyl_lysine.AllCell.bed ...

  2. File list: Oth.Adp.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Adipocyte htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.10.Crotonyl_lysine.AllCell.bed ...

  3. File list: Oth.ALL.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine All cell type...s SRX1060566,SRX1060567,SRX1060557 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.ALL.20.Crotonyl_lysine.AllCell.bed ...

  4. File list: Oth.Epd.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Epd.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Epidermis htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Epd.20.Crotonyl_lysine.AllCell.bed ...

  5. File list: Oth.Epd.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Epd.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Epidermis htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Epd.50.Crotonyl_lysine.AllCell.bed ...

  6. File list: Oth.ALL.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine All cell type...s SRX1060567,SRX1060566,SRX1060557 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.ALL.10.Crotonyl_lysine.AllCell.bed ...

  7. File list: Oth.Adp.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Adipocyte htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.05.Crotonyl_lysine.AllCell.bed ...

  8. File list: Oth.Dig.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Dig.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Digestive tra...ct http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Dig.10.Crotonyl_lysine.AllCell.bed ...

  9. File list: Oth.EmF.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.EmF.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Embryonic fib...roblast http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.EmF.50.Crotonyl_lysine.AllCell.bed ...

  10. File list: Oth.CDV.50.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.CDV.50.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Cardiovascula...r http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.CDV.50.Crotonyl_lysine.AllCell.bed ...

  11. File list: Oth.Gon.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Gon.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Gonad SRX1060...566,SRX1060567,SRX1060557 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Gon.05.Crotonyl_lysine.AllCell.bed ...

  12. File list: Oth.NoD.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.NoD.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine No descriptio...n http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.NoD.10.Crotonyl_lysine.AllCell.bed ...

  13. File list: Oth.PSC.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Pluripotent s...tem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.05.Crotonyl_lysine.AllCell.bed ...

  14. File list: Oth.Epd.10.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Epd.10.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Epidermis htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Epd.10.Crotonyl_lysine.AllCell.bed ...

  15. File list: Oth.ALL.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine All cell type...s SRX1060566,SRX1060567,SRX1060557 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.ALL.05.Crotonyl_lysine.AllCell.bed ...

  16. File list: Oth.PSC.20.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.PSC.20.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Pluripotent s...tem cell http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.PSC.20.Crotonyl_lysine.AllCell.bed ...

  17. File list: Oth.Epd.05.Crotonyl_lysine.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Epd.05.Crotonyl_lysine.AllCell mm9 TFs and others Crotonyl lysine Epidermis htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Epd.05.Crotonyl_lysine.AllCell.bed ...

  18. File list: His.Oth.50.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Oth.50.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation O...thers http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Oth.50.Pan_lysine_crotonylation.AllCell.bed ...

  19. File list: His.Bon.20.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Bon.20.Pan_lysine_crotonylation.AllCell hg19 Histone Pan lysine crotonylation B...one http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Bon.20.Pan_lysine_crotonylation.AllCell.bed ...

  20. File list: His.Plc.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Plc.10.Pan_lysine_crotonylation.AllCell mm9 Histone Pan lysine crotonylation Pl...acenta http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Plc.10.Pan_lysine_crotonylation.AllCell.bed ...