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Sample records for a-kinase anchoring protein

  1. A-Kinase Anchoring Protein Mediates TRPV1 Thermal Hyperalgesia through PKA Phosphorylation of TRPV1

    Jeske, Nathaniel A.; Diogenes, Anibal; Ruparel, Nikita B.; Fehrenbacher, Jill C.; Henry, Michael; Akopian, Armen N.; Hargreaves, Kenneth M.

    2008-01-01

    Certain phosphorylation events are tightly controlled by scaffolding proteins such as A-Kinase Anchoring Protein (AKAP). On nociceptive terminals, phosphorylation of transient receptor potential channel type 1 (TRPV1) results in the sensitization to many different stimuli, contributing to the development of hyperalgesia. In this study, we investigated the functional involvement of AKAP150 in mediating sensitization of TRPV1, and found that AKAP150 is co-expressed in trigeminal ganglia (TG) ne...

  2. Emerging roles of A-kinase anchoring proteins in cardiovascular pathophysiology.

    Diviani, Dario; Reggi, Erica; Arambasic, Miroslav; Caso, Stefania; Maric, Darko

    2016-07-01

    Heart and blood vessels ensure adequate perfusion of peripheral organs with blood and nutrients. Alteration of the homeostatic functions of the cardiovascular system can cause hypertension, atherosclerosis, and coronary artery disease leading to heart injury and failure. A-kinase anchoring proteins (AKAPs) constitute a family of scaffolding proteins that are crucially involved in modulating the function of the cardiovascular system both under physiological and pathological conditions. AKAPs assemble multifunctional signaling complexes that ensure correct targeting of the cAMP-dependent protein kinase (PKA) as well as other signaling enzymes to precise subcellular compartments. This allows local regulation of specific effector proteins that control the function of vascular and cardiac cells. This review will focus on recent advances illustrating the role of AKAPs in cardiovascular pathophysiology. The accent will be mainly placed on the molecular events linked to the control of vascular integrity and blood pressure as well as on the cardiac remodeling process associated with heart failure. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Integration of Developmental and Environmental Cues in the Heart edited by Marcus Schaub and Hughes Abriel. PMID:26643253

  3. Optic atrophy 1 is an A-kinase anchoring protein on lipid droplets that mediates adrenergic control of lipolysis.

    Pidoux, Guillaume; Witczak, Oliwia; Jarnæss, Elisabeth; Myrvold, Linda; Urlaub, Henning; Stokka, Anne Jorunn; Küntziger, Thomas Michel; Tasken, Kjetil

    2011-01-01

    Adrenergic stimulation of adipoytes induces PKA-dependent phosphorylation of perilipin, the major regulator of lipolysis in lipid droplets. The mitochondrial dynamics regulator Optic Atrophy 1 is the A-kinase anchoring protein on lipid droplets, and is required for the regulation of lipolysis.

  4. Molecular evolution of a-kinase anchoring protein (AKAP-7: implications in comparative PKA compartmentalization

    Johnson Keven R

    2012-07-01

    Full Text Available Abstract Background A-Kinase Anchoring Proteins (AKAPs are molecular scaffolding proteins mediating the assembly of multi-protein complexes containing cAMP-dependent protein kinase A (PKA, directing the kinase in discrete subcellular locations. Splice variants from the AKAP7 gene (AKAP15/18 are vital components of neuronal and cardiac phosphatase complexes, ion channels, cardiac Ca2+ handling and renal water transport. Results Shown in evolutionary analyses, the formation of the AKAP7-RI/RII binding domain (required for AKAP/PKA-R interaction corresponds to vertebrate-specific gene duplication events in the PKA-RI/RII subunits. Species analyses of AKAP7 splice variants shows the ancestral AKAP7 splice variant is AKAP7α, while the ancestral long form AKAP7 splice variant is AKAP7γ. Multi-species AKAP7 gene alignments, show the recent formation of AKAP7δ occurs with the loss of native AKAP7γ in rats and basal primates. AKAP7 gene alignments and two dimensional Western analyses indicate that AKAP7γ is produced from an internal translation-start site that is present in the AKAP7δ cDNA of mice and humans but absent in rats. Immunofluorescence analysis of AKAP7 protein localization in both rat and mouse heart suggests AKAP7γ replaces AKAP7δ at the cardiac sarcoplasmic reticulum in species other than rat. DNA sequencing identified Human AKAP7δ insertion-deletions (indels that promote the production of AKAP7γ instead of AKAP7δ. Conclusions This AKAP7 molecular evolution study shows that these vital scaffolding proteins developed in ancestral vertebrates and that independent mutations in the AKAP7 genes of rodents and early primates has resulted in the recent formation of AKAP7δ, a splice variant of likely lesser importance in humans than currently described.

  5. A-kinase-anchoring proteins coordinate inflammatory responses to cigarette smoke in airway smooth muscle.

    Poppinga, Wilfred J; Heijink, Irene H; Holtzer, Laura J; Skroblin, Philipp; Klussmann, Enno; Halayko, Andrew J; Timens, Wim; Maarsingh, Harm; Schmidt, Martina

    2015-04-15

    β2-Agonist inhibitors can relieve chronic obstructive pulmonary disease (COPD) symptoms by stimulating cyclic AMP (cAMP) signaling. A-kinase-anchoring proteins (AKAPs) compartmentalize cAMP signaling by establishing protein complexes. We previously reported that the β2-agonist fenoterol, direct activation of protein kinase A (PKA), and exchange factor directly activated by cAMP decrease cigarette smoke extract (CSE)-induced release of neutrophil attractant interleukin-8 (IL-8) from human airway smooth muscle (ASM) cells. In the present study, we tested the role of AKAPs in CSE-induced IL-8 release from ASM cells and assessed the effect of CSE on the expression levels of different AKAPs. We also studied mRNA and protein expression of AKAPs in lung tissue from patients with COPD. Our data show that CSE exposure of ASM cells decreases AKAP5 and AKAP12, both capable of interacting with β2-adrenoceptors. In lung tissue of patients with COPD, mRNA levels of AKAP5 and AKAP12 were decreased compared with lung tissue from controls. Using immunohistochemistry, we detected less AKAP5 protein in ASM of patients with COPD Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage II compared with control subjects. St-Ht31, which disrupts AKAP-PKA interactions, augmented CSE-induced IL-8 release from ASM cells and diminished its suppression by fenoterol, an effect mediated by disturbed ERK signaling. The modulatory role of AKAP-PKA interactions in the anti-inflammatory effects of fenoterol in ASM cells and the decrease in expression of AKAP5 and AKAP12 in response to cigarette smoke and in lungs of patients with COPD suggest that cigarette smoke-induced changes in AKAP5 and AKAP12 in patients with COPD may affect efficacy of pharmacotherapy. PMID:25637608

  6. Compartmentalization Role of A-Kinase Anchoring Proteins (AKAPs in Mediating Protein Kinase A (PKA Signaling and Cardiomyocyte Hypertrophy

    Abeer Rababa'h

    2014-12-01

    Full Text Available The Beta-adrenergic receptors (β-ARs stimulation enhances contractility through protein kinase-A (PKA substrate phosphorylation. This PKA signaling is conferred in part by PKA binding to A-kinase anchoring proteins (AKAPs. AKAPs coordinate multi-protein signaling networks that are targeted to specific intracellular locations, resulting in the localization of enzyme activity and transmitting intracellular actions of neurotransmitters and hormones to its target substrates. In particular, mAKAP (muscle-selective AKAP has been shown to be present on the nuclear envelope of cardiomyocytes with various proteins including: PKA-regulatory subunit (RIIα, phosphodiesterase-4D3, protein phosphatase-2A, and ryanodine receptor (RyR2. Therefore, through the coordination of spatial-temporal signaling of proteins and enzymes, mAKAP controls cyclic-adenosine monophosphate (cAMP levels very tightly and functions as a regulator of PKA-mediated substrate phosphorylation leading to changes in calcium availability and myofilament calcium sensitivity. The goal of this review is to elucidate the critical compartmentalization role of mAKAP in mediating PKA signaling and regulating cardiomyocyte hypertrophy by acting as a scaffolding protein. Based on our literature search and studying the structure–function relationship between AKAP scaffolding protein and its binding partners, we propose possible explanations for the mechanism by which mAKAP promotes cardiac hypertrophy.

  7. Myomegalin is a novel A-kinase anchoring protein involved in the phosphorylation of cardiac myosin binding protein C

    Riedemann Johann

    2011-05-01

    Full Text Available Abstract Background Cardiac contractility is regulated by dynamic phosphorylation of sarcomeric proteins by kinases such as cAMP-activated protein kinase A (PKA. Efficient phosphorylation requires that PKA be anchored close to its targets by A-kinase anchoring proteins (AKAPs. Cardiac Myosin Binding Protein-C (cMyBPC and cardiac troponin I (cTNI are hypertrophic cardiomyopathy (HCM-causing sarcomeric proteins which regulate contractility in response to PKA phosphorylation. Results During a yeast 2-hybrid (Y2H library screen using a trisphosphorylation mimic of the C1-C2 region of cMyBPC, we identified isoform 4 of myomegalin (MMGL as an interactor of this N-terminal cMyBPC region. As MMGL has previously been shown to interact with phosphodiesterase 4D, we speculated that it may be a PKA-anchoring protein (AKAP. To investigate this possibility, we assessed the ability of MMGL isoform 4 to interact with PKA regulatory subunits R1A and R2A using Y2H-based direct protein-protein interaction assays. Additionally, to further elucidate the function of MMGL, we used it as bait to screen a cardiac cDNA library. Other PKA targets, viz. CARP, COMMD4, ENO1, ENO3 and cTNI were identified as putative interactors, with cTNI being the most frequent interactor. We further assessed and confirmed these interactions by fluorescent 3D-co-localization in differentiated H9C2 cells as well as by in vivo co-immunoprecipitation. We also showed that quantitatively more interaction occurs between MMGL and cTNI under β-adrenergic stress. Moreover, siRNA-mediated knockdown of MMGL leads to reduction of cMyBPC levels under conditions of adrenergic stress, indicating that MMGL-assisted phosphorylation is requisite for protection of cMyBPC against proteolytic cleavage. Conclusions This study ascribes a novel function to MMGL isoform 4: it meets all criteria for classification as an AKAP, and we show that is involved in the phosphorylation of cMyBPC as well as cTNI, hence MMGL

  8. Activation of NMDA receptors leads to phosphorylation of TRPV1 S800 by protein kinase C and A-Kinase anchoring protein 150 in rat trigeminal ganglia

    Lee, Jongseok; Chung, Man-Kyo; Ro, Jin Y.

    2012-01-01

    A-Kinase anchoring protein 150 (AKAP150) is required for the phosphorylation of transient receptor potential cation channel subfamily V member 1 (TRPV1) by PKA or PKC in sensory neurons and, hence, affects TRPV1-dependent hyperalgesia under pathological conditions. Recently, we showed that the activation of N-methyl-d-aspartate (NMDA) receptors sensitizes TRPV1 by enhancing serine phosphorylation through PKC in trigeminal nociceptors. In this study, we extended this observation by investigati...

  9. Scaffolding during the cell cycle by A-kinase anchoring proteins

    Han, B; Poppinga, W J; Schmidt, M

    2015-01-01

    Cell division relies on coordinated regulation of the cell cycle. A process including a well-defined series of strictly regulated molecular mechanisms involving cyclin-dependent kinases, retinoblastoma protein, and polo-like kinases. Dysfunctions in cell cycle regulation are associated with disease

  10. Scaffolding during the cell cycle by A-kinase anchoring proteins

    Han, B.; Poppinga, W J; Schmidt, M.

    2015-01-01

    Cell division relies on coordinated regulation of the cell cycle. A process including a well-defined series of strictly regulated molecular mechanisms involving cyclin-dependent kinases, retinoblastoma protein, and polo-like kinases. Dysfunctions in cell cycle regulation are associated with disease such as cancer, diabetes, and neurodegeneration. Compartmentalization of cellular signaling is a common strategy used to ensure the accuracy and efficiency of cellular responses. Compartmentalizati...

  11. Topology of cAMP dependent protein kinase and A-kinase anchor proteins in mammalian mitochondria

    Nuzzi, R.; Sardanelli, A. M.; Dobrová, Zuzana; Signorile, A.; De Rasmo, D.; Papa, S.

    Milano, 2005, s. 57. ISSN 0021-2938. [SIB 2005. Riccione (IT), 27.09.2005-30.09.2005] Institutional research plan: CEZ:AV0Z50200510 Keywords : protein kinase * mammalian mitochondria Subject RIV: EE - Microbiology, Virology

  12. Identification of cAMP-dependent protein kinase holoenzymes in preantral- and preovulatory-follicle-enriched ovaries, and their association with A-kinase-anchoring proteins.

    Carr, D W; Cutler, R E; Cottom, J E; Salvador, L M; Fraser, I D; Scott, J D; Hunzicker-Dunn, M

    1999-12-01

    Undifferentiated cells from preantral (PA) follicles respond to high levels of cAMP in a different manner than do differentiated cells from preovulatory (PO) follicles. We hypothesized that this differential response of PA and PO cells to cAMP could be due, in part, to either a difference in the profile of isoforms that comprise the cAMP-dependent protein kinase (PKA) holoenzymes and/or a difference in the interaction of PKA with A-kinase-anchoring proteins (AKAPs). To test these hypotheses, PKA activity, PKA holoenzymes, PKA subunits and AKAPs from PA and PO ovaries were compared. Soluble PKA holoenzymes and regulatory (R) subunits were separated by DEAE-cellulose chromatography and sucrose-density-gradient centrifugation. PKA R subunits were distinguished by photoaffinity labelling, autophosphorylation, size, isoelectric point and immunoreactivity. AKAPs were identified by RII subunit overlay assays and immunoreactivity. The results showed that extracts from PA and PO ovaries exhibited equivalent PKA holoenzyme profiles and activities, characterized by low levels of PKA type I (PKAI) holoenzyme and two distinct PKAII holoenzyme peaks, one containing only RIIbeta subunits (PKAIIbeta) and one containing both PKAIIbeta and PKAIIalpha holoenzymes. Both PA and PO ovarian extracts also contained PKA catalytic (C)-subunit-free RIalpha, while only PO ovaries exhibited C-subunit-free RIIbeta. Consistent with the elevated levels of C-subunit-free RIIbeta in PO cells, PKA activation in PO cells required higher concentrations of forskolin than that in PA cells. While extracts of PA and PO ovaries exhibited a number of similar AKAPs, including four prominent ones reactive with anti-AKAP-KL antisera (where AKAP-KL is an AKAP especially abundant in kidney and liver), cAMP-agarose affinity chromatography revealed two major differences in AKAP binding to purified R subunits. PO ovaries contained increased levels of AKAP80 (AKAP of 80 kDa) bound selectively to R subunits in DEAE

  13. Occurrence of A-kinase anchor protein and associated cAMP-dependent protein kinase in the inner compartment of mammalian mitochondria

    Sardanelli, A. M.; Signorile, A.; Nuzzi, R.; De Rasmo, D.; Dobrová, Zuzana; Drahota, Zdeněk; Occhiello, A.; Pica, A.; Papa, S.

    2006-01-01

    Roč. 580, č. 24 (2006), s. 5690-5696. ISSN 0014-5793 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z50110509 Keywords : pka * akap proteins * cardiomyocytes Subject RIV: EE - Microbiology, Virology Impact factor: 3.372, year: 2006

  14. How curved membranes recruit amphipathic helices and protein anchoring motifs

    Hatzakis, Nikos; Bhatia, Vikram Kjøller; Larsen, Jannik;

    2009-01-01

    : membrane-anchored proteins. The fact that unrelated structural motifs such as alpha-helices and alkyl chains sense MC led us to propose that MC sensing is a generic property of curved membranes rather than a property of the anchoring molecules. We therefore anticipate that MC will promote the...... redistribution of proteins that are anchored in membranes through other types of hydrophobic moieties....

  15. FragAnchor: A Large-Scale Predictor of Glycosylphosphatidylinositol Anchors in Eukaryote Protein Sequences by Qualitative Scoring

    2007-01-01

    A glycosylphosphatidylinositol (GPI) anchor is a common but complex C-terminal post-translational modification of extracellular proteins in eukaryotes. Here we investigate the problem of correctly annotating GPI-anchored proteins for the growing number of sequences in public databases. We developed a computational system, called FragAnchor, based on the tandem use of a neural network (NN) and a hidden Markov model (HMM). Firstly, NN selects potential GPI-anchored proteins in a dataset, then HMM parses these potential GPI signals and refines the prediction by qualitative scoring. FragAnchor correctly predicted 91% of all the GPI-anchored proteins annotated in the Swiss-Prot database.In a large-scale analysis of 29 eukaryote proteomes, FragAnchor predicted that the percentage of highly probable GPI-anchored proteins is between 0.21% and 2.01%. The distinctive feature of FragAnchor, compared with other systems,is that it targets only the C-terminus of a protein, making it less sensitive to the background noise found in databases and possible incomplete protein sequences. Moreover, FragAnchor can be used to predict GPI-anchored proteins in all eukaryotes. Finally, by using qualitative scoring, the predictions combine both sensitivity and information content. The predictor is publicly available at http: // navet. ics. hawaii.edu/~fraganchor/NNHMM/NNHMM.html.

  16. Proteomic analysis of GPI-anchored membrane proteins

    Jung, Hye Ryung; Jensen, Ole Nørregaard

    2006-01-01

    humans. GPI-APs confer important cellular functions as receptors, enzymes and scaffolding molecules. Specific enzymes and detergent extraction methods combined with separation technologies and mass spectrometry permit proteomic analysis of GPI-APs from plasma membrane preparations to reveal cell......Glycosyl-phosphatidyl-inositol-anchored proteins (GPI-APs) represent a subset of post-translationally modified proteins that are tethered to the outer leaflet of the plasma membrane via a C-terminal GPI anchor. GPI-APs are found in a variety of eukaryote species, from pathogenic microorganisms to...

  17. Addition of lipid substituents of mammalian protein glycosylphosphoinositol anchors.

    Singh, N.; Zoeller, R A; Tykocinski, M. L.; Lazarow, P B; Tartakoff, A M

    1994-01-01

    A single metabolic path leading to synthesis of ether lipids is known in animal cells, the major products of which are plasmalogens. To learn whether this peroxisomal path is also responsible for the synthesis of base-resistant lipid components of glycosylphosphoinositol (GPI)-anchored membrane proteins, we have investigated the structure of anchor precursor mannolipids both in wild-type cells (CHO-K1 and a macrophage-like line, RAW 264.7) and in two corresponding mutant cells in which ether ...

  18. Proteomic analysis of glycosylphosphatidylinositol-anchored membrane proteins

    Elortza, Felix; Nühse, Thomas S; Foster, Leonard J;

    2003-01-01

    Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are a functionally and structurally diverse family of post-translationally modified membrane proteins found mostly in the outer leaflet of the plasma membrane in a variety of eukaryotic cells. Although the general role of GPI-APs remains...... unclear, they have attracted attention because they act as enzymes and receptors in cell adhesion, differentiation, and host-pathogen interactions. GPI-APs may represent potential diagnostic and therapeutic targets in humans and are interesting in plant biotechnology because of their key role in root...... and 44 GPI-APs in an Arabidopsis thaliana membrane preparation, representing the largest experimental dataset of GPI-anchored proteins to date....

  19. The glycosylphosphatidylinositol-anchored protein repertoire of babesia bovis and its significance for erythrocyte invasion

    Glycosylphosphatidyl-anchored proteins are particularly abundant on the surface of pathogenic protozoans and might play an important role for parasite survival. In the present work the relevance of GPI-anchored proteins for erythrocyte invasion of Babesia bovis, one of the tick-transmitted causative...

  20. GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane

    Sevcsik, Eva; Brameshuber, Mario; Fölser, Martin; Weghuber, Julian; Honigmann, Alf; Schütz, Gerhard J.

    2015-01-01

    The organization of proteins and lipids in the plasma membrane has been subject of a long-lasting debate. Membrane rafts of higher lipid chain order were proposed to mediate protein interactions, but have thus far not been directly observed. Here, we use protein micropatterning combined with single-molecule tracking to put current models to the test: we rearranged lipid-anchored raft proteins (glycosylphosphatidylinositol(GPI)-anchored mGFP) directly in the live cell plasma membrane and measu...

  1. GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane.

    Sevcsik, E.; Brameshuber, M.; Fölser, M.; Weghuber, J.; Honigmann, A.; Schütz, G

    2015-01-01

    The organization of proteins and lipids in the plasma membrane has been the subject of a long-lasting debate. Membrane rafts of higher lipid chain order were proposed to mediate protein interactions, but have thus far not been directly observed. Here we use protein micropatterning combined with single-molecule tracking to put current models to the test: we rearranged lipid-anchored raft proteins (glycosylphosphatidylinositol(GPI)-anchored-mGFP) directly in the live cell plasma membrane and me...

  2. Insulin-induced decrease in protein phosphorylation in rat adipocytes not explained by decreased A-kinase activity

    In isolated rat adipocytes, insulin inhibits lipolysis to a greater extent than would be predicted by the decrease in (-/+)cAMP activity ratio of cAMP-dependent protein kinase [A-kinase], from which it was speculated that insulin promotes the dephosphorylation of hormone-sensitive lipase. They have examined the phosphorylation state of cellular proteins under conditions of varying A-kinase activities in the presence and absence of insulin. Protein phosphorylation was determined by SDS-PAGE electrophoresis of extracts from 32P-loaded cells; glycerol and A-kinase activity ratios were measured in the cytosolic extracts from control, non-radioactive cells. Increased protein phosphorylation in general occurred over the same range of A-kinase activity ratios, 0.1-0.3, associated with increased glycerol release. The insulin-induced decrease in lipolysis was associated with a decrease in the 32P content of several proteins, an effect not explained by the modest reduction in A-kinase activity by insulin. This effect of insulin on protein phosphorylation was lost as the A-kinase activity ratios exceeded 0.5. The results suggest that insulin promotes the dephosphorylation of those adipocyte proteins which are subject to phosphorylation by A-kinase

  3. Insulin-induced decrease in protein phosphorylation in rat adipocytes not explained by decreased A-kinase activity

    Egan, J.J.; Greenberg, A.S.; Chang, M.K.; Londos, C.

    1987-05-01

    In isolated rat adipocytes, insulin inhibits lipolysis to a greater extent than would be predicted by the decrease in (-/+)cAMP activity ratio of cAMP-dependent protein kinase (A-kinase), from which it was speculated that insulin promotes the dephosphorylation of hormone-sensitive lipase. They have examined the phosphorylation state of cellular proteins under conditions of varying A-kinase activities in the presence and absence of insulin. Protein phosphorylation was determined by SDS-PAGE electrophoresis of extracts from /sup 32/P-loaded cells; glycerol and A-kinase activity ratios were measured in the cytosolic extracts from control, non-radioactive cells. Increased protein phosphorylation in general occurred over the same range of A-kinase activity ratios, 0.1-0.3, associated with increased glycerol release. The insulin-induced decrease in lipolysis was associated with a decrease in the /sup 32/P content of several proteins, an effect not explained by the modest reduction in A-kinase activity by insulin. This effect of insulin on protein phosphorylation was lost as the A-kinase activity ratios exceeded 0.5. The results suggest that insulin promotes the dephosphorylation of those adipocyte proteins which are subject to phosphorylation by A-kinase.

  4. Cholesterol-dependent retention of GPI-anchored proteins in endosomes.

    Mayor, S; Sabharanjak, S; Maxfield, F R

    1998-01-01

    Several cell surface eukaryotic proteins have a glycosylphosphatidylinositol (GPI) modification at the Cterminal end that serves as their sole means of membrane anchoring. Using fluorescently labeled ligands and digital fluorescence microscopy, we show that contrary to the potocytosis model, GPI-anchored proteins are internalized into endosomes that contain markers for both receptor-mediated uptake (e.g. transferrin) and fluid phase endocytosis (e.g. dextrans). This was confirmed by immunogol...

  5. An accessory protein required for anchoring and assembly of amyloid fibres in B. subtilis biofilms.

    Romero, Diego; Vlamakis, Hera; Losick, Richard; Kolter, Roberto

    2011-06-01

    Cells within Bacillus subtilis biofilms are held in place by an extracellular matrix that contains cell-anchored amyloid fibres, composed of the amyloidogenic protein TasA. As biofilms age they disassemble because the cells release the amyloid fibres. This release appears to be the consequence of incorporation of D-tyrosine, D-leucine, D-tryptophan and D-methionine into the cell wall. Here, we characterize the in vivo roles of an accessory protein TapA (TasA anchoring/assembly protein; previously YqxM) that serves both to anchor the fibres to the cell wall and to assemble TasA into fibres. TapA is found in discrete foci in the cell envelope and these foci disappear when cells are treated with a mixture of D-amino acids. Purified cell wall sacculi retain a functional form of this anchoring protein such that purified fibres can be anchored to the sacculi in vitro. In addition, we show that TapA is essential for the proper assembly of the fibres. Its absence results in a dramatic reduction in TasA levels and what little TasA is left produces only thin fibres that are not anchored to the cell. PMID:21477127

  6. An Accessory Protein Required for Anchoring and Assembly of Amyloid Fibers in B. subtilis Biofilms

    Romero, Diego; Vlamakis, Hera; Losick, Richard; Kolter, Roberto

    2011-01-01

    Cells within Bacillus subtilis biofilms are held in place by an extracellular matrix that contains cell-anchored amyloid fibers, composed of the amyloidogenic protein TasA. As biofilms age they disassemble because the cells release the amyloid fibers. This release appears to be the consequence of incorporation of D-tyrosine, D-leucine, D-tryptophan and D-methionine into the cell wall. Here, we characterize the in vivo roles of an accessory protein TapA (TasA anchoring/assembly protein; previo...

  7. Distinct Pathways Mediate the Sorting of Tail-anchored Mitochondrial Outer Membrane Proteins

    Little is known about the biogenesis of tail-anchored (TA) proteins localized to the mitochondrial outer membrane in plant cells. To address this issue, we screened all of the (>600) known and predicted TA proteins in Arabidopsis thaliana for those annotated, based on Gene Ontology, to possess mitoc...

  8. Peptide nanofibers modified with a protein by using designed anchor molecules bearing hydrophobic and functional moieties.

    Miyachi, Ayaka; Takahashi, Tsuyoshi; Matsumura, Sachiko; Mihara, Hisakazu

    2010-06-11

    Self-assembly of peptides and proteins is a key feature of biological functions. Short amphiphilic peptides designed with a beta-sheet structure can form sophisticated nanofiber structures, and the fibers are available as nanomaterials for arranging biomolecules. Peptide FI (H-PKFKIIEFEP-OH) self-assembles into nanofibers with a coiled fine structure, as reported in our previous work. We have constructed anchor molecules that have both a binding moiety for the fiber structure and a functional unit capable of capturing target molecules, with the purpose of arranging proteins on the designed peptide nanofibers. Designed anchors containing an alkyl chain as a binding unit and biotin as a functional moiety were found to bind to peptide fibers FI and F2i (H-ALEAKFAAFEAKLA-NH(2)). The surface-exposed biotin moiety on the fibers could capture an anti-biotin antibody. Moreover, hydrophobic dipeptide anchor units composed of iminodiacetate connected to Phe-Phe or Ile-Ile and a peptide composed of six histidine residues connected to biotin could also connect FI peptide fibers to the anti-biotin antibody through the chelation of Ni(2+) ions. This strategy of using designed anchors opens a novel approach to constructing nanoscale protein arrays on peptide nanomaterials. PMID:20419712

  9. Crucial Role for Prion Protein Membrane Anchoring in the Neuroinvasion and Neural Spread of Prion Infection ▿

    Klingeborn, Mikael; Race, Brent; Meade-White, Kimberly D.; Rosenke, Rebecca; Striebel, James F.; Chesebro, Bruce

    2010-01-01

    In nature prion diseases are usually transmitted by extracerebral prion infection, but clinical disease results only after invasion of the central nervous system (CNS). Prion protein (PrP), a host-encoded glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein, is necessary for prion infection and disease. Here, we investigated the role of the anchoring of PrP on prion neuroinvasion by studying various inoculation routes in mice expressing either anchored or anchorless PrP. In contr...

  10. A glycolipid-anchored prion protein is endocytosed via clathrin-coated pits

    1994-01-01

    The cellular prion protein (PrPc) is a glycolipid-anchored, cell surface protein of unknown function, a posttranslationally modified isoform of which PrPSc is involved in the pathogenesis of Creutzfeldt- Jakob disease, scrapie, and other spongiform encephalopathies. We have shown previously that chPrP, a chicken homologue of mammalian PrPC, constitutively cycles between the cell surface and an endocytic compartment, with a transit time of approximately 60 min in cultured neuroblastoma cells. ...

  11. Phospholipase D specific for the phosphatidylinositol anchor of cell-surface proteins is abundant in plasma

    An enzyme activity capable of degrading the glycosyl-phosphatidylinositol membrane anchor of cell-surface proteins has previously been reported in a number of mammalian tissues. The experiments reported here demonstrate that this anchor-degrading activity is also abundant in mammalian plasma. The activity was inhibited by EGTA or 1,10-phenanthroline. It was capable of removing the anchor from alkaline phosphatase, 5'-nucleotidase, and variant surface glycoprotein but had little or no activity toward phosphatidylinositol or phosphatidylcholine. Phosphatidic acid was the only 3H-labeled product when this enzyme hydrolyzed [3H]myristate-labeled variant surface glycoprotein. It could be distinguished from the Ca2=-dependent inositol phospholipid-specific phospholipase C activity in several rat tissues on the basis of its molecular size and its sensitivity to 1,10-phenanthroline. The data therefore suggest that this activity is due to a phospholipase D with specificity for glycosylphosphatidylinositol structures. Although the precise physiological function of this anchor-specific phospholipase D remains to be determined, these findings indicate that it could play an important role in regulating the expression and release of cell-surface proteins in vivo

  12. Lipid transfer proteins do their thing anchored at membrane contact sites… but what is their thing?

    Wong, Louise H; Levine, Tim P

    2016-04-15

    Membrane contact sites are structures where two organelles come close together to regulate flow of material and information between them. One type of inter-organelle communication is lipid exchange, which must occur for membrane maintenance and in response to environmental and cellular stimuli. Soluble lipid transfer proteins have been extensively studied, but additional families of transfer proteins have been identified that are anchored into membranes by transmembrane helices so that they cannot diffuse through the cytosol to deliver lipids. If such proteins target membrane contact sites they may be major players in lipid metabolism. The eukaryotic family of so-called Lipid transfer proteins Anchored at Membrane contact sites (LAMs) all contain both a sterol-specific lipid transfer domain in the StARkin superfamily (related to StART/Bet_v1), and one or more transmembrane helices anchoring them in the endoplasmic reticulum (ER), making them interesting subjects for study in relation to sterol metabolism. They target a variety of membrane contact sites, including newly described contacts between organelles that were already known to make contact by other means. Lam1-4p target punctate ER-plasma membrane contacts. Lam5p and Lam6p target multiple contacts including a new category: vacuolar non-NVJ cytoplasmic ER (VancE) contacts. These developments confirm previous observations on tubular lipid-binding proteins (TULIPs) that established the importance of membrane anchored proteins for lipid traffic. However, the question remaining to be solved is the most difficult of all: are LAMs transporters, or alternately are they regulators that affect traffic more indirectly? PMID:27068964

  13. Is hexamerin receptor a GPI-anchored protein in Achaea janata (Lepidoptera: Noctuidae)?

    Madhusudhan Budatha; Thuirei Jacob Ningshen; Aparna Dutta-Gupta

    2011-08-01

    The process of uptake of hexamerins during metamorphosis from insect haemolymph by fat body cells is reminiscent of receptor-mediated endocytosis. Previously, we had identified a hexamerin-binding protein (HBP) and reported for the first time that uptake of hexamerins is dependent on the phosphorylation of HBP partly by a tyrosine kinase, which is, in turn, activated by 20-hydroxyecdysone (20E). However, the exact nature of HBP and the mechanism of interaction are still unknown. Here we report the possibility of HBP being a GPI-anchored protein in the fat body of Achaea janata and its role in the tyrosine-kinase-mediated phosphorylation signalling. Digestion of fat body membrane preparation with bacterial phosphatidylinositol-specific phospholipase C (PI-PLC), and the subsequent recognition by antibodies specific for the cross-reacting determinant (CRD), revealed that HBP is glycosylphosphatidylinositol (GPI)-anchored protein and, further, that the hexamerin binding to HBP was inhibited after digestion. Hexamerin overlay assay (HOA) of co-immunoprecipitated in vitro phosphorylated HBP showed exclusive binding to ∼120 kDa protein. Lectin-binding analysis of hexamerins revealed the presence of -acetylgalactosamine (GalNAc) and -acetylglucosamine (GluNAc), whereas HBP showed the presence of GalNac alone. Mild chemical deglycosylation studies and binding interaction in the presence of sugars revealed that glycan moieties are possibly not involved in the interaction between HBP and hexamerins. Taken together, these results suggest that HBP may be a GPI-anchored protein, and interaction and activation of HBP is through lipid-linked non-receptor src tyrosine kinases. However, additional studies are needed to prove that HBP is a GPI-anchored protein.

  14. The cholesterol membrane anchor of the Hedgehog protein confers stable membrane association to lipid-modified proteins

    Peters, Carsten; Wolf, Alexander; Wagner, Melanie; Kuhlmann, Jürgen; Waldmann, Herbert

    2004-01-01

    The Hedgehog proteins are potent organizers of animal development. They carry a cholesterol ester at the C terminus of their signaling domain. The membrane anchoring mediated by this lipophilic modification was studied by means of an approach integrating cell biology, biochemistry, biophysics, and organic chemistry techniques. Sterol-modified and fluorescent-labeled Hedgehog-derived peptides and proteins were synthesized and investigated in biophysical and cell-biological assays. These experi...

  15. Listeria monocytogenes phosphatidylinositol (PI)-specific phospholipase C has low activity on glycosyl-PI-anchored proteins.

    Gandhi, A J; Perussia, B; Goldfine, H

    1993-01-01

    The ability of the phosphatidylinositol-specific phospholipase C (PI-PLC) from Listeria monocytogenes to hydrolyze glycosyl phosphatidylinositol (GPI)-anchored membrane proteins was compared with the ability of the PI-PLC from Bacillus thuringiensis to hydrolyze such proteins. The L. monocytogenes enzyme produced no detectable release of acetylcholinesterase from bovine, sheep, and human erythrocytes. The cleavage of the GPI anchors of alkaline phosphatase from rat and rabbit kidney slices wa...

  16. Distinct patterns of spread of prion infection in brains of mice expressing anchorless or anchored forms of prion protein

    Rangel, Alejandra; Race, Brent; Phillips, Katie; Striebel, James; Kurtz, Nancy; Chesebro, Bruce

    2014-01-01

    Background In humans and animals, prion protein (PrP) is usually expressed as a glycophosphatidylinositol (GPI)-anchored membrane protein, but anchorless PrP may be pathogenic in humans with certain familial prion diseases. Anchored PrP expressed on neurons mediates spread of prions along axons in the peripheral and central nervous systems. However, the mechanism of prion spread in individuals expressing anchorless PrP is poorly understood. Here we studied prion spread within brain of mice ex...

  17. GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane

    Sevcsik, Eva; Brameshuber, Mario; Fölser, Martin; Weghuber, Julian; Honigmann, Alf; Schütz, Gerhard J.

    2015-04-01

    The organization of proteins and lipids in the plasma membrane has been the subject of a long-lasting debate. Membrane rafts of higher lipid chain order were proposed to mediate protein interactions, but have thus far not been directly observed. Here we use protein micropatterning combined with single-molecule tracking to put current models to the test: we rearranged lipid-anchored raft proteins (glycosylphosphatidylinositol(GPI)-anchored-mGFP) directly in the live cell plasma membrane and measured the effect on the local membrane environment. Intriguingly, this treatment does neither nucleate the formation of an ordered membrane phase nor result in any enrichment of nanoscopic-ordered domains within the micropatterned regions. In contrast, we find that immobilized mGFP-GPIs behave as inert obstacles to the diffusion of other membrane constituents without influencing their membrane environment over distances beyond their physical size. Our results indicate that phase partitioning is not a fundamental element of protein organization in the plasma membrane.

  18. Comparison of cell wall proteins of Saccharomyces cerevisiae as anchors for cell surface expression of heterologous proteins.

    van der Vaart, J. M.; te Biesebeke, R; Chapman, J.W.; Toschka, H Y; Klis, F M; Verrips, C. T.

    1997-01-01

    The carboxyl-terminal regions of five cell wall proteins (Cwp1p, Cwp2p, Ag alpha 1p, Tip1p, and Flo1p) and three potential cell wall proteins (Sed1p, YCR89w, and Tir1p) all proved capable of immobilizing alpha-galactosidase in the cell wall of Saccharomyces cerevisiae. The fraction of the total amount of fusion protein that was localized to the cell wall varied depending on the anchor domain used. The highest proportion of cell wall incorporation was achieved with Cwp2p, Ag alpha 1p, or Sed1p...

  19. Sialic Acid within the Glycosylphosphatidylinositol Anchor Targets the Cellular Prion Protein to Synapses.

    Bate, Clive; Nolan, William; McHale-Owen, Harriet; Williams, Alun

    2016-08-12

    Although the cellular prion protein (PrP(C)) is concentrated at synapses, the factors that target PrP(C) to synapses are not understood. Here we demonstrate that exogenous PrP(C) was rapidly targeted to synapses in recipient neurons derived from Prnp knock-out((0/0)) mice. The targeting of PrP(C) to synapses was dependent upon both neuronal cholesterol concentrations and the lipid and glycan composition of its glycosylphosphatidylinositol (GPI) anchor. Thus, the removal of either an acyl chain or sialic acid from the GPI anchor reduced the targeting of PrP(C) to synapses. Isolated GPIs (derived from PrP(C)) were also targeted to synapses, as was IgG conjugated to these GPIs. The removal of sialic acid from GPIs prevented the targeting of either the isolated GPIs or the IgG-GPI conjugate to synapses. Competition studies showed that pretreatment with sialylated GPIs prevented the targeting of PrP(C) to synapses. These results are consistent with the hypothesis that the sialylated GPI anchor attached to PrP(C) acts as a synapse homing signal. PMID:27325697

  20. Die Klonierung und Charakterisierung des protein kinase A anchoring protein r(rat)Ht31 aus der Rattenniere

    Pepperle, Barbara

    2010-01-01

    In an attempt to isolate protein kinase A anchoring proteins (AKAPs) involved in vasopressin-mediated water reabsorption a partial cDNA clone with an open reading frame of 582 amino acids (rHt31 (GenBank accession number AF387102) and 67,7 % identity to the human AKAP hHt31 was isolated. One RII binding site was mapped to amino acid residues 65-78; a second binding site with lower RII affinity was mapped to amino acid residues 470-576. The complete cDNA sequence of hHt31 was determined by a c...

  1. A prokaryotic membrane anchor sequence: carboxyl terminus of bacteriophage f1 gene III protein retains it in the membrane.

    Boeke, J D; Model, P

    1982-01-01

    Gene III protein of bacteriophage f1 is inserted into the host cell membrane where it is assembled into phage particles. A truncated form of gene III protein, encoded by a recombinant plasmid and lacking the carboxyl terminus, does not remain in the membrane but instead appears to slip through it. Fusion of a hydrophobic "membrane anchor" from another membrane protein, the gene VIII protein, to the truncated gene III protein (by manipulation of the recombinant plasmid) restores membrane ancho...

  2. Comparison of cell wall proteins of Saccharomyces cerevisiae as anchors for cell surface expression of heterologous proteins.

    Van der Vaart, J M; te Biesebeke, R; Chapman, J W; Toschka, H Y; Klis, F M; Verrips, C T

    1997-01-01

    The carboxyl-terminal regions of five cell wall proteins (Cwp1p, Cwp2p, Ag alpha 1p, Tip1p, and Flo1p) and three potential cell wall proteins (Sed1p, YCR89w, and Tir1p) all proved capable of immobilizing alpha-galactosidase in the cell wall of Saccharomyces cerevisiae. The fraction of the total amount of fusion protein that was localized to the cell wall varied depending on the anchor domain used. The highest proportion of cell wall incorporation was achieved with Cwp2p, Ag alpha 1p, or Sed1p as an anchor. Although 80% of these fusion proteins were incorporated in the cell wall, the total production of alpha-galactosidase-Ag alpha 1p was sixfold lower than that of alpha-galactosidase-Cwp2p and eightfold lower than that of alpha-galactosidase-Sed1p. Differences in mRNA levels were not responsible for this discrepancy, nor was an intracellular accumulation of alpha-galactosidase-Ag alpha 1p detectable. A lower translation efficiency of the alpha-galactosidase-AG alpha 1 fusion construct is most likely to be responsible for the low level of protein production. alpha-Galactosidase immobilized by the carboxyl-terminal 67 amino acids of Cwp2p was most effective in the hydrolysis of the high-molecular-weight substrate guar gum from Cyamopsis tetragonoloba. This indicates that the use of a large anchoring domain does not necessarily result in a better exposure of the immobilized enzyme to the exterior of the yeast cell. PMID:9023939

  3. The mechanism of membrane-associated steps in tail-anchored protein insertion

    Mariappan, Malaiyalam; Mateja, Agnieszka; Dobosz, Malgorzata; Bove, Elia; Hegde, Ramanujan S.; Keenan, Robert J. (NIH); (UC)

    2012-06-19

    Tail-anchored (TA) membrane proteins destined for the endoplasmic reticulum are chaperoned by cytosolic targeting factors that deliver them to a membrane receptor for insertion. Although a basic framework for TA protein recognition is now emerging, the decisive targeting and membrane insertion steps are not understood. Here we reconstitute the TA protein insertion cycle with purified components, present crystal structures of key complexes between these components and perform mutational analyses based on the structures. We show that a committed targeting complex, formed by a TA protein bound to the chaperone ATPase Get3, is initially recruited to the membrane through an interaction with Get2. Once the targeting complex has been recruited, Get1 interacts with Get3 to drive TA protein release in an ATPase-dependent reaction. After releasing its TA protein cargo, the now-vacant Get3 recycles back to the cytosol concomitant with ATP binding. This work provides a detailed structural and mechanistic framework for the minimal TA protein insertion cycle.

  4. The role of Listeria monocytogenes cell wall surface anchor protein LapB in virulence, adherence, and intracellular replication

    Lmof2365_2117 is a Listeria monocytogenes putative cell wall surface anchor protein with a conserved domain found in collagen binding proteins. We constructed a deletion mutation in lmof2365_2117 in serotype 4b strain F2365, evaluated its virulence, and determined its ability to adhere and invade co...

  5. Characterisation of a cell wall-anchored protein of Staphylococcus saprophyticus associated with linoleic acid resistance

    King Nathan P

    2012-01-01

    Full Text Available Abstract Background The Gram-positive bacterium Staphylococcus saprophyticus is the second most frequent causative agent of community-acquired urinary tract infections (UTI, accounting for up to 20% of cases. A common feature of staphylococci is colonisation of the human skin. This involves survival against innate immune defenses including antibacterial unsaturated free fatty acids such as linoleic acid which act by disrupting bacterial cell membranes. Indeed, S. saprophyticus UTI is usually preceded by perineal skin colonisation. Results In this study we identified a previously undescribed 73.5 kDa cell wall-anchored protein of S. saprophyticus, encoded on plasmid pSSAP2 of strain MS1146, which we termed S. saprophyticus surface protein F (SssF. The sssF gene is highly prevalent in S. saprophyticus clinical isolates and we demonstrate that the SssF protein is expressed at the cell surface. However, unlike all other characterised cell wall-anchored proteins of S. saprophyticus, we were unable to demonstrate a role for SssF in adhesion. SssF shares moderate sequence identity to a surface protein of Staphylococcus aureus (SasF recently shown to be an important mediator of linoleic acid resistance. Using a heterologous complementation approach in a S. aureus sasF null genetic background, we demonstrate that SssF is associated with resistance to linoleic acid. We also show that S. saprophyticus strains lacking sssF are more sensitive to linoleic acid than those that possess it. Every staphylococcal genome sequenced to date encodes SssF and SasF homologues. Proteins in this family share similar predicted secondary structures consisting almost exclusively of α-helices in a probable coiled-coil formation. Conclusions Our data indicate that SssF is a newly described and highly prevalent surface-localised protein of S. saprophyticus that contributes to resistance against the antibacterial effects of linoleic acid. SssF is a member of a protein family

  6. A unifying mechanism accounts for sensing of membrane curvature by BAR domains, amphipathic helices and membrane-anchored proteins

    Bhatia, Vikram Kjøller; Hatzakis, Nikos; Stamou, Dimitrios

    2010-01-01

    itself. We thus anticipate that membrane curvature will promote the redistribution of proteins that are anchored in membranes through any type of hydrophobic moiety, a thesis that broadens tremendously the implications of membrane curvature for protein sorting, trafficking and signaling in cell biology.......The discovery of proteins that recognize membrane curvature created a paradigm shift by suggesting that membrane shape may act as a cue for protein localization that is independent of lipid or protein composition. Here we review recent data on membrane curvature sensing by three structurally...... unrelated motifs: BAR domains, amphipathic helices and membrane-anchored proteins. We discuss the conclusion that the curvature of the BAR dimer is not responsible for sensing and that the sensing properties of all three motifs can be rationalized by the physicochemical properties of the curved membrane...

  7. QM/MD studies of the dynamics of the MTSL spin label in Aurora-A kinase protein activation loop

    Concilio, Maria Grazia; Bayliss, Richard; Burgess, Selena

    2015-01-01

    Molecular dynamics(MD)simulations using a graphics processing unit (GPU) has been employed in order to determine the conformational space of the methane-thiosulfonate spin label (MTSL) attached to the activation loop of the Aurora-A kinase protein and compared with quantum mechanical (QM) methods rooted on density functional theory (DFT). MD provided a wealth of information about interactions between the MTSL and the residues of the protein and on the different motional contributions to the overall dynamics of the MTSL. Data obtained from MD were seen to be in good agreement with those obtained from QM but the dynamics of the system revealed more interactions than those observed from QM methods. A strong correlation between the tumbling of the protein and the transitions of the X4 and X5 dihedral angles of the MTSL, was observed with a consequent effect also the distribution of the nitroxide(NO)group in the space. Theoretical EPR spectra calculated from opportunely selected MD frames showing interactions betw...

  8. Targeting of a Tail-anchored Protein to Endoplasmic Reticulum and Mitochondrial Outer Membrane by Independent but Competing Pathways

    Borgese, Nica; Gazzoni, Ilaria; Barberi, Massimo; Colombo, Sara; Pedrazzini, Emanuela

    2001-01-01

    Many mitochondrial outer membrane (MOM) proteins have a transmembrane domain near the C terminus and an N-terminal cytosolic moiety. It is not clear how these tail-anchored (TA) proteins posttranslationally select their target, but C-terminal charged residues play an important role. To investigate how discrimination between MOM and endoplasmic reticulum (ER) occurs, we used mammalian cytochrome b5, a TA protein existing in two, MOM or ER localized, versions. Substi...

  9. Distinct pathways mediate the sorting of tail-anchored proteins to the plastid outer envelope.

    Preetinder K Dhanoa

    Full Text Available BACKGROUND: Tail-anchored (TA proteins are a distinct class of membrane proteins that are sorted post-translationally to various organelles and function in a number of important cellular processes, including redox reactions, vesicular trafficking and protein translocation. While the molecular targeting signals and pathways responsible for sorting TA proteins to their correct intracellular destinations in yeasts and mammals have begun to be characterized, relatively little is known about TA protein biogenesis in plant cells, especially for those sorted to the plastid outer envelope. METHODOLOGY/PRINCIPAL FINDINGS: Here we investigated the biogenesis of three plastid TA proteins, including the 33-kDa and 34-kDa GTPases of the translocon at the outer envelope of chloroplasts (Toc33 and Toc34 and a novel 9-kDa protein of unknown function that we define here as an outer envelope TA protein (OEP9. Using a combination of in vivo and in vitro assays we show that OEP9 utilizes a different sorting pathway than that used by Toc33 and Toc34. For instance, while all three TA proteins interact with the cytosolic OEP chaperone/receptor, AKR2A, the plastid targeting information within OEP9 is distinct from that within Toc33 and Toc34. Toc33 and Toc34 also appear to differ from OEP9 in that their insertion is dependent on themselves and the unique lipid composition of the plastid outer envelope. By contrast, the insertion of OEP9 into the plastid outer envelope occurs in a proteinaceous-dependent, but Toc33/34-independent manner and membrane lipids appear to serve primarily to facilitate normal thermodynamic integration of this TA protein. CONCLUSIONS/SIGNIFICANCE: Collectively, the results provide evidence in support of at least two sorting pathways for plastid TA outer envelope proteins and shed light on not only the complex diversity of pathways involved in the targeting and insertion of proteins into plastids, but also the molecular mechanisms that underlie

  10. ADAM12 is expressed in the tumour vasculature and mediates ectodomain shedding of several membrane-anchored endothelial proteins

    Frohlich, Camilla; Klitgaard, Marie; Noer, Julie B;

    2013-01-01

    ADAM (a disintegrin and metalloproteinase) 12 is a metalloprotease implicated in cancer progression. ADAM12 can activate membrane-anchored proteins, such as sonic hedgehog, Delta-like 1 and certain epidermal growth factor receptor ligands, through a process called ectodomain shedding. We screened...

  11. Functional convergence of signalling by GPI-anchored and anchorless forms of a salamander protein implicated in limb regeneration.

    Blassberg, R. A.; Garza-Garcia, A.; Janmohamed, A.; Gates, P. B.; Brockes, J. P.

    2011-01-01

    The GPI-anchor is an established determinant of molecular localisation and various functional roles have been attributed to it. The newt GPI-anchored three-finger protein (TFP) Prod1 is an important regulator of cell behaviour during limb regeneration, but it is unclear how it signals to the interior of the cell. Prod1 was expressed by transfection in cultured newt limb cells and activated transcription and expression of matrix metalloproteinase 9 (MMP9) by a pathway involving ligand-independ...

  12. Restrictive glycosylphosphatidylinositol anchor synthesis in cwh6/gpi3 yeast cells causes aberrant biogenesis of cell wall proteins.

    Vossen, J.H.; Müller, W. H.; Lipke, P N; Klis, F. M.

    1997-01-01

    We previously reported that the defects in the Saccharomyces cerevisiae cwh6 Calcofluor white-hypersensitive cell wall mutant are caused by a mutation in SPT14/GPI3, a gene involved in glycosylphosphatidylinositol (GPI) anchor biosynthesis. Here we describe the effect of cwh6/spt14/gpi3 on the biogenesis of cell wall proteins. It was found that the release of precursors of cell wall proteins from the endoplasmic reticulum (ER) was retarded. This was accompanied by proliferation of ER structur...

  13. Legionella pneumophila exploits PI(4P to anchor secreted effector proteins to the replicative vacuole.

    2006-05-01

    Full Text Available The causative agent of Legionnaires' disease, Legionella pneumophila, employs the intracellular multiplication (Icm/defective organelle trafficking (Dot type IV secretion system (T4SS to upregulate phagocytosis and to establish a replicative vacuole in amoebae and macrophages. Legionella-containing vacuoles (LCVs do not fuse with endosomes but recruit early secretory vesicles. Here we analyze the role of host cell phosphoinositide (PI metabolism during uptake and intracellular replication of L. pneumophila. Genetic and pharmacological evidence suggests that class I phosphatidylinositol(3 kinases (PI3Ks are dispensable for phagocytosis of wild-type L. pneumophila but inhibit intracellular replication of the bacteria and participate in the modulation of the LCV. Uptake and degradation of an icmT mutant strain lacking a functional Icm/Dot transporter was promoted by PI3Ks. We identified Icm/Dot-secreted proteins which specifically bind to phosphatidylinositol(4 phosphate (PI(4P in vitro and preferentially localize to LCVs in the absence of functional PI3Ks. PI(4P was found to be present on LCVs using as a probe either an antibody against PI(4P or the PH domain of the PI(4P-binding protein FAPP1 (phosphatidylinositol(4 phosphate adaptor protein-1. Moreover, the presence of PI(4P on LCVs required a functional Icm/Dot T4SS. Our results indicate that L. pneumophila modulates host cell PI metabolism and exploits the Golgi lipid second messenger PI(4P to anchor secreted effector proteins to the LCV.

  14. PIG-S and PIG-T, essential for GPI anchor attachment to proteins, form a complex with GAA1 and GPI8

    Ohishi, Kazuhito; Inoue, Norimitsu; Kinoshita, Taroh

    2001-01-01

    Many eukaryotic cell surface proteins are anchored to the plasma membrane via glycosylphosphatidylinositol (GPI). The GPI transamidase mediates GPI anchoring in the endoplasmic reticulum, by replacing a protein’s C-terminal GPI attachment signal peptide with a pre-assembled GPI. During this transamidation reaction, the GPI transamidase forms a carbonyl intermediate with a substrate protein. It was known that the GPI transamidase is a complex containing GAA1 and GPI8. Here, we report two new c...

  15. Fatal transmissible amyloid encephalopathy: a new type of prion disease associated with lack of prion protein membrane anchoring.

    Bruce Chesebro

    2010-03-01

    Full Text Available Prion diseases are fatal neurodegenerative diseases of humans and animals characterized by gray matter spongiosis and accumulation of aggregated, misfolded, protease-resistant prion protein (PrPres. PrPres can be deposited in brain in an amyloid-form and/or non-amyloid form, and is derived from host-encoded protease-sensitive PrP (PrPsen, a protein normally anchored to the plasma membrane by glycosylphosphatidylinositol (GPI. Previously, using heterozygous transgenic mice expressing only anchorless PrP, we found that PrP anchoring to the cell membrane was required for typical clinical scrapie. However, in the present experiments, using homozygous transgenic mice expressing two-fold more anchorless PrP, scrapie infection induced a new fatal disease with unique clinical signs and altered neuropathology, compared to non-transgenic mice expressing only anchored PrP. Brain tissue of transgenic mice had high amounts of infectivity, and histopathology showed dense amyloid PrPres plaque deposits without gray matter spongiosis. In contrast, infected non-transgenic mice had diffuse non-amyloid PrPres deposits with significant gray matter spongiosis. Brain graft studies suggested that anchored PrPsen expression was required for gray matter spongiosis during prion infection. Furthermore, electron and light microscopic studies in infected transgenic mice demonstrated several pathogenic processes not seen in typical prion disease, including cerebral amyloid angiopathy and ultrastructural alterations in perivascular neuropil. These findings were similar to certain human familial prion diseases as well as to non-prion human neurodegenerative diseases, such as Alzheimer's disease.

  16. Localization in rat heart mitochondria of outer and inner pools of camp dependent protein kinase, a kinase anchor proteins and PP2C serine phosphatase

    Sardanalli, A. M.; Dobrová, Zuzana; Signorile, A.; Nuzzi, R.; Drahota, Z.; Papa, S.

    Milano, 2003, s. 250. [Join Symposia with the British Biochemical Society /SIP 2003./. Milano (IT), 15.09.2003-18.09.2003] Institutional research plan: CEZ:AV0Z5020903 Keywords : pcamp * akap * aqdq Subject RIV: EE - Microbiology, Virology

  17. Anchor reinforcements

    Levkovich, P.Ye.; Sal' nikov, V.K.; Savich, N.S.

    1980-11-30

    An anchor reinforcement includes an anchor shaft, an elastic jig with a separator wall having compartments placed parallely along it and filled with reinforcement material and a device for destroying the jig wall separator. To quickly put the anchor in place and increase the reliability of the reinforcement by mixing the reinforcement material components better, the device for destroying the jig separator wall was made in the shape of a shovel, fastened to the anchor shaft and the separator wall has a longitudinal hollow for holding the anchor shaft/shovel.

  18. Molecular insights into DNA binding and anchoring by the Bacillus subtilis sporulation kinetochore-like RacA protein

    Schumacher, Maria A.; Lee, Jeehyun; Zeng, Wenjie

    2016-01-01

    During Bacillus subtilis sporulation, segregating sister chromosomes are anchored to cell poles and the chromosome is remodeled into an elongated structure called the axial filament. Data indicate that a developmentally regulated protein called RacA is involved in these functions. To gain insight into how RacA performs these diverse processes we performed a battery of structural and biochemical analyses. These studies show that RacA contains an N-terminal winged-helix-turn-helix module connected by a disordered region to a predicted coiled-coil domain. Structures capture RacA binding the DNA using distinct protein–protein interfaces and employing adjustable DNA docking modes. This unique DNA binding mechanism indicates how RacA can both specifically recognize its GC-rich centromere and also non-specifically bind the DNA. Adjacent RacA molecules within the protein–DNA structure interact leading to DNA compaction, suggesting a mechanism for axial filament formation. We also show that the RacA C-domain coiled coil directly contacts the coiled coil region of the polar protein DivIVA, which anchors RacA and hence the chromosome to the pole. Thus, our combined data reveal unique DNA binding properties by RacA and provide insight into the DNA remodeling and polar anchorage functions of the protein. PMID:27085804

  19. Anchoring secreted proteins in endoplasmic reticulum by plant oleosin: the example of vitamin B12 cellular sequestration by transcobalamin.

    Laurent Pons

    Full Text Available BACKGROUND: Oleosin is a plant protein localized to lipid droplets and endoplasmic reticulum of plant cells. Our idea was to use it to target functional secretory proteins of interest to the cytosolic side of the endoplasmic reticulum of mammalian cells, through expressing oleosin-containing chimeras. We have designed this approach to create cellular models deficient in vitamin B12 (cobalamin because of the known problematics associated to the obtainment of effective vitamin B12 deficient cell models. This was achieved by the overexpression of transcobalamin inside cells through anchoring to oleosin. METHODOLOGY: chimera gene constructs including transcobalamin-oleosin (TC-O, green fluorescent protein-transcobalamin-oleosin (GFP-TC-O and oleosin-transcobalamin (O-TC were inserted into pAcSG2 and pCDNA3 vectors for expression in sf9 insect cells, Caco2 (colon carcinoma, NIE-115 (mouse neuroblastoma, HEK (human embryonic kidney, COS-7 (Green Monkey SV40-transfected kidney fibroblasts and CHO (Chinese hamster ovary cells. The subcellular localization, the changes in vitamin B12 binding activity and the metabolic consequences were investigated in both Caco2 and NIE-115 cells. PRINCIPAL FINDINGS: vitamin B12 binding was dramatically higher in TC-O than that in O-TC and wild type (WT. The expression of GFP-TC-O was observed in all cell lines and found to be co-localized with an ER-targeted red fluorescent protein and calreticulin of the endoplasmic reticulum in Caco2 and COS-7 cells. The overexpression of TC-O led to B12 deficiency, evidenced by impaired conversion of cyano-cobalamin to ado-cobalamin and methyl-cobalamin, decreased methionine synthase activity and reduced S-adenosyl methionine to S-adenosyl homocysteine ratio, as well as increases in homocysteine and methylmalonic acid concentration. CONCLUSIONS/SIGNIFICANCE: the heterologous expression of TC-O in mammalian cells can be used as an effective strategy for investigating the cellular

  20. A tail-anchored myotonic dystrophy protein kinase isoform induces perinuclear clustering of mitochondria, autophagy, and apoptosis.

    Ralph J A Oude Ophuis

    Full Text Available BACKGROUND: Studies on the myotonic dystrophy protein kinase (DMPK gene and gene products have thus far mainly concentrated on the fate of length mutation in the (CTGn repeat at the DNA level and consequences of repeat expansion at the RNA level in DM1 patients and disease models. Surprisingly little is known about the function of DMPK protein products. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrate here that transient expression of one major protein product of the human gene, the hDMPK A isoform with a long tail anchor, results in mitochondrial fragmentation and clustering in the perinuclear region. Clustering occurred in a variety of cell types and was enhanced by an intact tubulin cytoskeleton. In addition to morphomechanical changes, hDMPK A expression induces physiological changes like loss of mitochondrial membrane potential, increased autophagy activity, and leakage of cytochrome c from the mitochondrial intermembrane space accompanied by apoptosis. Truncation analysis using YFP-hDMPK A fusion constructs revealed that the protein's tail domain was necessary and sufficient to evoke mitochondrial clustering behavior. CONCLUSION/SIGNIFICANCE: Our data suggest that the expression level of the DMPK A isoform needs to be tightly controlled in cells where the hDMPK gene is expressed. We speculate that aberrant splice isoform expression might be a codetermining factor in manifestation of specific DM1 features in patients.

  1. Intermolecular Interaction between Anchoring Subunits Specify Subcellular Targeting and Function of RGS Proteins in Retina ON-Bipolar Neurons.

    Sarria, Ignacio; Orlandi, Cesare; McCall, Maureen A; Gregg, Ronald G; Martemyanov, Kirill A

    2016-03-01

    In vertebrate retina, light responses generated by the rod photoreceptors are transmitted to the second-order neurons, the ON-bipolar cells (ON-BC), and this communication is indispensible for vision in dim light. In ON-BCs, synaptic transmission is initiated by the metabotropic glutamate receptor, mGluR6, that signals via the G-protein Go to control opening of the effector ion channel, TRPM1. A key role in this process belongs to the GTPase Activating Protein (GAP) complex that catalyzes Go inactivation upon light-induced suppression of glutamate release in rod photoreceptors, thereby driving ON-BC depolarization to changes in synaptic input. The GAP complex has a striking molecular complexity. It contains two Regulator of G-protein Signaling (RGS) proteins RGS7 and RGS11 that directly act on Go and two adaptor subunits: RGS Anchor Protein (R9AP) and the orphan receptor, GPR179. Here we examined the organizational principles of the GAP complex in ON-BCs. Biochemical experiments revealed that RGS7 binds to a conserved site in GPR179 and that RGS11 in vivo forms a complex only with R9AP. R9AP and GPR179 are further integrated via direct protein-protein interactions involving their cytoplasmic domains. Elimination of GPR179 prevents postsynaptic accumulation of R9AP. Furthermore, concurrent knock-out of both R9AP and RGS7 does not reconfigure the GAP complex and completely abolishes synaptic transmission, resulting in a novel mouse model of night blindness. Based on these results, we propose a model of hierarchical assembly and function of the GAP complex that supports ON-BCs visual signaling. PMID:26961947

  2. Lipid storage in large and small rat adipocytes by vesicle-associated glycosylphosphatidylinositol-anchored proteins.

    Müller, Günter; Wied, Susanne; Dearey, Elisabeth-Ann; Wetekam, Eva-Maria; Biemer-Daub, Gabriele

    2010-01-01

    Adipose tissue mass in mammals expands by increasing the average cell volume and/or total number of the adipocytes. Upregulated lipid storage in fully differentiated adipocytes resulting in their enlargement is well documented and thought to be a critical mechanism for the expansion of adipose tissue depots during the growth of both lean and obese animals and human beings. A novel molecular mechanism for the regulation of lipid storage and cell size in rat adipocytes was recently elucidated for the physiological stimuli, palmitate and H(2)O(2), and the antidiabetic sulfonylurea drug, glimepiride. It encompasses (1) the release of small vesicles, so-called adiposomes, harboring the glycosylphosphatidylinositol -anchored (c)AMP-degrading phosphodiesterase Gce1 and 5'-nucleotidase CD73 from donor adipocytes, (2) the transfer of the adiposomes and their interaction with detergent-insoluble glycolipid-enriched microdomains of the plasma membrane of acceptor adipocytes, (3) the translocation of Gce1 and CD73 from the adiposomes to the intracellular lipid droplets of the acceptor adipocytes, and (4) the degradation of (c)AMP at the lipid droplet surface zone by Gce1 and CD73 in the acceptor adipocytes, leading to the upregulation of the esterification of fatty acids into triacylglycerol s and the downregulation of their release from triacylglycerols. This mechanism may provide novel strategies for the therapy of metabolic diseases, such as type 2 diabetes and obesity. PMID:20865369

  3. The Novel Tail-anchored Membrane Protein Mff Controls Mitochondrial and Peroxisomal Fission in Mammalian Cells

    Gandre-Babbe, Shilpa; van der Bliek, Alexander M.

    2008-01-01

    Few components of the mitochondrial fission machinery are known, even though mitochondrial fission is a complex process of vital importance for cell growth and survival. Here, we describe a novel protein that controls mitochondrial fission. This protein was identified in a small interfering RNA (siRNA) screen using Drosophila cells. The human homologue of this protein was named Mitochondrial fission factor (Mff). Mitochondria of cells transfected with Mff siRNA form a closed network similar t...

  4. ‘Fractional Recovery’ Analysis of a Presynaptic Synaptotagmin 1-Anchored Endocytic Protein Complex

    Khanna, Rajesh; Li, Qi; Stanley, Elise F.

    2006-01-01

    Background The integral synaptic vesicle protein and putative calcium sensor, synaptotagmin 1 (STG), has also been implicated in synaptic vesicle (SV) recovery. However, proteins with which STG interacts during SV endocytosis remain poorly understood. We have isolated an STG-associated endocytic complex (SAE) from presynaptic nerve terminals and have used a novel fractional recovery (FR) assay based on electrostatic dissociation to identify SAE components and map the complex structure. The lo...

  5. Prenatal cocaine reduces AMPA receptor synaptic expression through hyperphosphorylation of the synaptic anchoring protein GRIP

    Bakshi, Kalindi; Gennaro, Serena; Chan, Christopher Y.; Kosciuk, Mary; Liu, Jingjing; Stucky, Andres; Trenkner, Ekkehart; FRIEDMAN, EITAN; Nagele, Robert G; Wang, Hoau-Yan

    2009-01-01

    Prenatal cocaine exposure produces sustained neurobehavioral and brain synaptic changes closely resembling those of animals with defective alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamatergic receptors (AMPARs). We hypothesized that prenatal cocaine exposure attenuates AMPAR signaling by interfering with AMPAR synaptic targeting. AMPAR function is governed by receptor cycling on and off the synaptic membrane through its interaction with GRIP, a PDZ domain protein that i...

  6. Expression pattern of GPI-anchored non-specific lipid transfer proteins in Physcomitrella patens

    Höglund, Andrey

    2011-01-01

    During the water-to-land transition, that occurred approximately 450 MYA, novel habitats wererevealed to the emerging plants. This terrestrial habitat was a harsh environment compared to theaquatic, with shifting substrate content, irregular supply of water, damaging UV-radiation andrapid fluctuating temperatures. Non-specific lipid transfer proteins (nsLTP) are today only foundin the land living plants and not in the green algae. This suggests that these genes might haveevolved to help the p...

  7. PbsP, a cell wall-anchored protein that binds plasminogen to promote hematogenous dissemination of group B Streptococcus.

    Buscetta, Marco; Firon, Arnaud; Pietrocola, Giampiero; Biondo, Carmelo; Mancuso, Giuseppe; Midiri, Angelina; Romeo, Letizia; Galbo, Roberta; Venza, Mario; Venza, Isabella; Kaminski, Pierre-Alexandre; Gominet, Myriam; Teti, Giuseppe; Speziale, Pietro; Trieu-Cuot, Patrick; Beninati, Concetta

    2016-07-01

    Streptococcus agalactiae (Group B Streptococcus or GBS) is a leading cause of invasive infections in neonates whose virulence is dependent on its ability to interact with cells and host components. We here characterized a surface protein with a critical function in GBS pathophysiology. This adhesin, designated PbsP, possesses two Streptococcal Surface Repeat domains, a methionine and lysine-rich region, and a LPXTG cell wall-anchoring motif. PbsP mediates plasminogen (Plg) binding both in vitro and in vivo and we showed that cell surface-bound Plg can be activated into plasmin by tissue plasminogen activator to increase the bacterial extracellular proteolytic activity. Absence of PbsP results in a decreased bacterial transmigration across brain endothelial cells and impaired virulence in a murine model of infection. PbsP is conserved among the main GBS lineages and is a major plasminogen adhesin in non-CC17 GBS strains. Importantly, immunization of mice with recombinant PbsP confers protective immunity. Our results indicate that GBS have evolved different strategies to recruit Plg which indicates that the ability to acquire cell surface proteolytic activity is essential for the invasiveness of this bacterium. PMID:26888569

  8. Inactivation of the srtA Gene in Streptococcus gordonii Inhibits Cell Wall Anchoring of Surface Proteins and Decreases In Vitro and In Vivo Adhesion

    Bolken, Tové C.; Franke, Christine A.; Jones, Kevin F.; Zeller, Gloria O.; Jones, C. Hal; Dutton, Emma K.; Hruby, Dennis E.

    2001-01-01

    The srtA gene product, SrtA, has been shown to be required for cell wall anchoring of protein A as well as virulence in the pathogenic bacterium Staphylococcus aureus. There are five major mechanisms for displaying proteins at the surface of gram-positive bacteria (P. Cossart and R. Jonquieres, Proc. Natl. Acad. Sci. USA 97:5013–5015, 2000). However, since many of the known surface proteins of gram-positive bacteria are believed to be exported and anchored via the sortase pathway, it was of interest to determine if srtA plays a similar role in other gram-positive bacteria. To that end, the srtA gene in the human oral commensal organism Streptococcus gordonii was insertionally inactivated. The srtA mutant S. gordonii exhibited a marked reduction in quantity of a specific anchored surface protein. Furthermore, the srtA mutant had reduced binding to immobilized human fibronectin and had a decreased ability to colonize the oral mucosa of mice. Taken together, these results suggest that the activity of SrtA plays an important role in the biology of nonpathogenic as well as pathogenic gram-positive cocci. PMID:11119491

  9. Display of active enzymes on the cell surface of Escherichia coli using PgsA anchor protein and their application to bioconversion.

    Narita, Junya; Okano, Kenji; Tateno, Toshihiro; Tanino, Takanori; Sewaki, Tomomitsu; Sung, Moon-Hee; Fukuda, Hideki; Kondo, Akihiko

    2006-05-01

    We have developed a novel Escherichia coli cell surface display system by employing PgsA as an anchoring motif. In our display system, C-terminal fusion to PgsA anchor protein from Bacillus subtilis was used. The enzymes selected for display were alpha-amylase (AmyA) from Streptococcus bovis 148 and lipase B (CALB) from Candida antarctica. The molecular mass values of AmyA and CALB are approximately 77 and 34 kDa, respectively. The enzymes were displayed on the surface as a fusion protein with a FLAG peptide tag at the C terminus. Both the PgsA-AmyA-FLAG and PgsA-CALB-FLAG fusion proteins were shown to be displayed by immunofluorescence labeling using anti-FLAG antibody. The displayed enzymes were active forms, and AmyA and CALB activities reached 990 U/g (dry cell weight) and 4.6 U/g (dry cell weight), respectively. AmyA-displaying E. coli cells grew utilizing cornstarch as the sole carbon source, while CALB-displaying E. coli cells catalyzed enantioselective transesterification, indicating that they are effective whole-cell biocatalysts. Since a target enzyme with a size of 77 kDa and an industrially useful lipase have been successfully displayed on the cell surface of E. coli for the first time, PgsA protein is probably a useful anchoring motif to display various enzymes. PMID:16133338

  10. Proteomic analysis of human norepinephrine transporter complexes reveals associations with protein phosphatase 2A anchoring subunit and 14-3-3 proteins

    The norepinephrine transporter (NET) terminates noradrenergic signals by clearing released NE at synapses. NET regulation by receptors and intracellular signaling pathways is supported by a growing list of associated proteins including syntaxin1A, protein phosphatase 2A (PP2A) catalytic subunit (PP2A-C), PICK1, and Hic-5. In the present study, we sought evidence for additional partnerships by mass spectrometry-based analysis of proteins co-immunoprecipitated with human NET (hNET) stably expressed in a mouse noradrenergic neuroblastoma cell line. Our initial proteomic analyses reveal multiple peptides derived from hNET, peptides arising from the mouse PP2A anchoring subunit (PP2A-Ar) and peptides derived from 14-3-3 proteins. We verified physical association of NET with PP2A-Ar via co-immunoprecipitation studies using mouse vas deferens extracts and with 14-3-3 via a fusion pull-down approach, implicating specifically the hNET NH2-terminus for interactions. The transporter complexes described likely support mechanisms regulating transporter activity, localization, and trafficking

  11. Comprehensive behavioral analysis of voltage-gated calcium channel beta-anchoring and -regulatory protein knockout mice

    Takafumi Miki

    2015-06-01

    Full Text Available Calcium (Ca2+ influx through voltage-gated Ca2+ channels (VGCCs induces numerous intracellular events such as neuronal excitability, neurotransmitter release, synaptic plasticity, and gene regulation. It has been shown that genes related to Ca2+ signaling, such as the CACNA1C, CACNB2, and CACNA1I genes that encode VGCC subunits, are associated with schizophrenia and other psychiatric disorders. Recently, VGCC beta-anchoring and -regulatory protein (BARP was identified as a novel regulator of VGCC activity via the interaction of VGCC β subunits. To examine the role of the BARP in higher brain functions, we generated BARP knockout (KO mice and conducted a comprehensive battery of behavioral tests. BARP KO mice exhibited greatly reduced locomotor activity, as evidenced by decreased vertical activity, stereotypic counts in the open field test, and activity level in the home cage, and longer latency to complete a session in spontaneous T-maze alteration test, which reached “study-wide significance”. Acoustic startle response was also reduced in the mutants. Interestingly, they showed multiple behavioral phenotypes that are seemingly opposite to those seen in the mouse models of schizophrenia and its related disorders, including increased working memory, flexibility, prepulse inhibition, and social interaction, and decreased locomotor activity, though many of these phenotypes are statistically weak and require further replications. These results demonstrate that BARP is involved in the regulation of locomotor activity and, possibly, emotionality. The possibility was also suggested that BARP KO mice may serve as a unique tool for investigating the pathogenesis/pathophysiology of schizophrenia and related disorders. Further evaluation of the molecular and physiological phenotypes of the mutant mice would provide new insights into the role of BARP in higher brain functions.

  12. Backbone conformational flexibility of the lipid modified membrane anchor of the human N-Ras protein investigated by solid-state NMR and molecular dynamics simulation.

    Vogel, Alexander; Reuther, Guido; Roark, Matthew B; Tan, Kui-Thong; Waldmann, Herbert; Feller, Scott E; Huster, Daniel

    2010-02-01

    The lipid modified human N-Ras protein, implicated in human cancer development, is of particular interest due to its membrane anchor that determines the activity and subcellular location of the protein. Previous solid-state NMR investigations indicated that this membrane anchor is highly dynamic, which may be indicative of backbone conformational flexibility. This article aims to address if a dynamic exchange between three structural models exist that had been determined previously. We applied a combination of solid-state nuclear magnetic resonance (NMR) methods and replica exchange molecular dynamics (MD) simulations using a Ras peptide that represents the terminal seven amino acids of the human N-Ras protein. Analysis of correlations between the conformations of individual amino acids revealed that Cys 181 and Met 182 undergo collective conformational exchange. Two major structures constituting about 60% of all conformations could be identified. The two conformations found in the simulation are in rapid exchange, which gives rise to low backbone order parameters and nuclear spin relaxation as measured by experimental NMR methods. These parameters were also determined from two 300 ns conventional MD simulations, providing very good agreement with the experimental data. PMID:19819220

  13. A phospholipid is the membrane-anchoring domain of a protein growth factor of molecular mass 34 kDa in placental trophoblasts

    Recently the authors isolated a protein growth factor of 34 kDa from trophoblastic membranes of human placenta. A fraction (∼50%) of the membrane-associated 34-kDa protein is peripherally associated--i.e., it can be released by high salt treatment. The remainder shows the characteristics of an integral membrane protein--i.e., its release requires detergent treatment. Here they report studies on the structural basis for membrane anchorage of the protein. Phospholipase C was found to release an immunoreactive 34-kDa polypeptide from intact isolated cytotrophoblasts. Studies with isolated trophoblastic membranes showed that phospholipase C specifically released the salt-resistant fraction of the 34-kDa polypeptide. The polypeptide released by phospholipase C showed the same electrophoretic mobility in NaDodSO4/PAGE as the polypeptide prior to phospholipase C treatment. The identity of the released protein with the 34-kDa growth factor has been established by both immunologic and receptor-binding assays. Other studies show that there is biosynthetic incorporation of [3H]myristate into the 34-kDa protein. The myristate label is labile to phospholipase C treatment. These results suggest that some of the 34-kDa protein is anchored to the plasma membrane via a posttranslationally added phospholipid. This mode of anchorage has been observed for some other membrane proteins and raises interesting questions regarding the role of this novel linkage in the mitogenic function of the 34-kDa polypeptide

  14. A 39-kD plasma membrane protein (IP39) is an anchor for the unusual membrane skeleton of Euglena gracilis

    The major integral plasma membrane protein (IP39) of Euglena gracilis was radiolabeled, peptide mapped, and dissected with proteases to identify cytoplasmic domains that bind and anchor proteins of the cell surface. When plasma membranes were radioiodinated and extracted with octyl glucoside, 98% of the extracted label was found in IP39 or the 68- and 110-kD oligomers of IP39. The octyl glucoside extracts were incubated with unlabeled cell surface proteins immobilized on nitrocellulose (overlays). Radiolabel from the membrane extract bound one (80 kD) of the two (80 and 86 kD) major membrane skeletal protein bands. Resolubilization of the bound label yielded a radiolabeled polypeptide identical in Mr to IP39. Intact plasma membranes were also digested with papain before or after radioiodination, thereby producing a cytoplasmically truncated IP39. The octyl glucoside extract of truncated IP39 no longer bound to the 80-kD membrane skeletal protein in the nitrocellulose overlays. EM of intact or trypsin digested plasma membranes incubated with membrane skeletal proteins under stringent conditions similar to those used in the nitrocellulose overlays revealed a partially reformed membrane skeletal layer. Little evidence of a membrane skeletal layer was found, however, when plasma membranes were predigested with papain before reassociation. A candidate 80-kD binding domain of IP39 has been tentatively identified as a peptide fragment that was present after trypsin digestion of plasma membranes, but was absent after papain digestion in two-dimensional peptide maps of IP39. Together, these data suggest that the unique peripheral membrane skeleton of Euglena binds to the plasma membrane through noncovalent interactions between the major 80-kD membrane skeletal protein and a small, papain sensitive cytoplasmic domain of IP39

  15. Roles of the Protruding Loop of Factor B Essential for the Localization of Lipoproteins (LolB) in the Anchoring of Bacterial Triacylated Proteins to the Outer Membrane*

    Hayashi, Yumi; Tsurumizu, Ryoji; Tsukahara, Jun; Takeda, Kazuki; Narita, Shin-ichiro; Mori, Makiko; Miki, Kunio; Tokuda, Hajime

    2014-01-01

    The Lol system comprising five Lol proteins, LolA through LolE, sorts Escherichia coli lipoproteins to outer membranes. The LolCDE complex, an ATP binding cassette transporter in inner membranes, releases outer membrane-specific lipoproteins in an ATP-dependent manner, causing formation of the LolA-lipoprotein complex in the periplasm. LolA transports lipoproteins through the periplasm to LolB on outer membranes. LolB is itself a lipoprotein anchored to outer membranes, although the membrane anchor is functionally dispensable. LolB then localizes lipoproteins to outer membranes through largely unknown mechanisms. The crystal structure of LolB is similar to that of LolA, and it possesses a hydrophobic cavity that accommodates acyl chains of lipoproteins. To elucidate the molecular function of LolB, a periplasmic version of LolB, mLolB, was mutagenized at various conserved residues. Despite the lack of acyl chains, most defective mutants were insoluble. However, a derivative with glutamate in place of leucine 68 was soluble and unable to localize lipoproteins to outer membranes. This leucine is present in a loop protruding from mLolB into an aqueous environment, and no analogous loop is present in LolA. Thus, leucine 68 was replaced with other residues. Replacement by acidic, but not hydrophobic, residues generated for the first time mLolB derivatives that can accept but cannot localize lipoproteins to outer membranes. Moreover, deletion of the leucine with neighboring residues impaired the lipoprotein receptor activity. Based on these observations, the roles of the protruding loop of LolB in the last step of lipoprotein sorting are discussed. PMID:24569999

  16. Roles of the protruding loop of factor B essential for the localization of lipoproteins (LolB) in the anchoring of bacterial triacylated proteins to the outer membrane.

    Hayashi, Yumi; Tsurumizu, Ryoji; Tsukahara, Jun; Takeda, Kazuki; Narita, Shin-ichiro; Mori, Makiko; Miki, Kunio; Tokuda, Hajime

    2014-04-11

    The Lol system comprising five Lol proteins, LolA through LolE, sorts Escherichia coli lipoproteins to outer membranes. The LolCDE complex, an ATP binding cassette transporter in inner membranes, releases outer membrane-specific lipoproteins in an ATP-dependent manner, causing formation of the LolA-lipoprotein complex in the periplasm. LolA transports lipoproteins through the periplasm to LolB on outer membranes. LolB is itself a lipoprotein anchored to outer membranes, although the membrane anchor is functionally dispensable. LolB then localizes lipoproteins to outer membranes through largely unknown mechanisms. The crystal structure of LolB is similar to that of LolA, and it possesses a hydrophobic cavity that accommodates acyl chains of lipoproteins. To elucidate the molecular function of LolB, a periplasmic version of LolB, mLolB, was mutagenized at various conserved residues. Despite the lack of acyl chains, most defective mutants were insoluble. However, a derivative with glutamate in place of leucine 68 was soluble and unable to localize lipoproteins to outer membranes. This leucine is present in a loop protruding from mLolB into an aqueous environment, and no analogous loop is present in LolA. Thus, leucine 68 was replaced with other residues. Replacement by acidic, but not hydrophobic, residues generated for the first time mLolB derivatives that can accept but cannot localize lipoproteins to outer membranes. Moreover, deletion of the leucine with neighboring residues impaired the lipoprotein receptor activity. Based on these observations, the roles of the protruding loop of LolB in the last step of lipoprotein sorting are discussed. PMID:24569999

  17. Control of lipid storage and cell size between adipocytes by vesicle-associated glycosylphosphatidylinositol-anchored proteins.

    Müller, Günter

    2011-02-01

    Adipose tissue mass in mammals is expanding by increasing the average cell volume as well as the total number of the adipocytes. Up-regulation of lipid storage in fully differentiated adipocytes resulting in their enlargement is well documented and thought to be a critical mechanism for the expansion of adipose tissue depots during the growth of both lean and obese animals and human beings. A novel molecular mechanism for the regulation of lipid storage and cell size in rat adipocytes has recently been elucidated for the physiological stimuli, palmitate and hydrogen peroxide, the anti-diabetic sulfonylurea drug, glimepiride, and insulin-mimetic phosphoinositolglycans. It encompasses (i) the release of small vesicles, so-called adiposomes, harbouring the glycosylphosphatidylinositol-anchored (c)AMP-degrading phosphodiesterase Gce1 and 5'-nuceotidase CD73 from large donor adipocytes, (ii) the transfer of the adiposomes and their interaction with detergent-insoluble glycolipid-enriched microdomains of the plasma membrane of small acceptor adipocytes, (iii) the translocation of Gce1 and CD73 from the adiposomes to the intracellular lipid droplets of the acceptor adipocytes and (iv) the degradation of (c)AMP at the lipid droplet surface zone by Gce1 and CD73 in the acceptor adipocytes. In concert, this sequence of events leads to up-regulation of esterification of fatty acids into triacylglycerol and down-regulation of their release from triacylglycerol. This apparent mechanism for shifting the triacylglycerol burden from large to small adipocytes may provide novel strategies for the therapy of metabolic diseases, such as type 2 diabetes and obesity. PMID:20883086

  18. Display of alpha-amylase on the surface of Lactobacillus casei cells by use of the PgsA anchor protein, and production of lactic acid from starch.

    Narita, Junya; Okano, Kenji; Kitao, Tomoe; Ishida, Saori; Sewaki, Tomomitsu; Sung, Moon-Hee; Fukuda, Hideki; Kondo, Akihiko

    2006-01-01

    We developed a new cell surface engineering system based on the PgsA anchor protein from Bacillus subtilis. In this system, the N terminus of the target protein was fused to the PgsA protein and the resulting fusion protein was expressed on the cell surface. Using this new system, we constructed a novel starch-degrading strain of Lactobacillus casei by genetically displaying alpha-amylase from the Streptococcus bovis strain 148 with a FLAG peptide tag (AmyAF). Localization of the PgsA-AmyA-FLAG fusion protein on the cell surface was confirmed by immunofluorescence microscopy and flow cytometric analysis. The lactic acid bacteria which displayed AmyAF showed significantly elevated hydrolytic activity toward soluble starch. By fermentation using AmyAF-displaying L. casei cells, 50 g/liter of soluble starch was reduced to 13.7 g/liter, and 21.8 g/liter of lactic acid was produced within about 24 h. The yield in terms of grams of lactic acid produced per gram of carbohydrate utilized was 0.60 g per g of carbohydrate consumed at 24 h. Since AmyA was immobilized on the cells, cells were recovered after fermentation and used repeatedly. During repeated utilization of cells, the lactic acid yield was improved to 0.81 g per g of carbohydrate consumed at 72 h. These results indicate that efficient simultaneous saccharification and fermentation from soluble starch to lactic acid were carried out by recombinant L. casei cells with cell surface display of AmyA. PMID:16391053

  19. Hypersensitivity to Suture Anchors

    Masafumi Goto

    2013-01-01

    Full Text Available Hypersensitivity to suture anchor is extremely rare. Herein, we present a case in which hypersensitivity to suture anchor was strongly suspected. The right rotator cuff of a 50-year-old woman was repaired with a metal suture anchor. Three weeks after the surgery, she developed erythema around her face, trunk, and hands, accompanied by itching. Infection was unlikely because no abnormalities were detected by blood testing or by medical examination. Suspicious of a metallic allergy, a dermatologist performed a patch testing 6 months after the first surgery. The patient had negative reactions to tests for titanium, aluminum, and vanadium, which were the principal components of the suture anchor. The anchor was removed 7 months after the first surgery, and the erythema disappeared immediately. When allergic symptoms occur and persist after the use of a metal anchor, removal should be considered as a treatment option even if the patch test result is negative.

  20. Susceptibility to anchoring effects

    Todd McElroy

    2007-02-01

    Full Text Available Previous research on anchoring has shown this heuristic to be a very robust psychological phenomenon ubiquitous across many domains of human judgment and decision-making. Despite the prevalence of anchoring effects, researchers have only recently begun to investigate the underlying factors responsible for how and in what ways a person is susceptible to them. This paper examines how one such factor, the Big-Five personality trait of openness-to-experience, influences the effect of previously presented anchors on participants' judgments. Our findings indicate that participants high in openness-to-experience were significantly more influenced by anchoring cues relative to participants low in this trait. These findings were consistent across two different types of anchoring tasks providing convergent evidence for our hypothesis.

  1. Identification and Characterization of a Novel Issatchenkia orientalis GPI-Anchored Protein, IoGas1, Required for Resistance to Low pH and Salt Stress.

    Matsushika, Akinori; Negi, Kanako; Suzuki, Toshihiro; Goshima, Tetsuya; Hoshino, Tamotsu

    2016-01-01

    The use of yeasts tolerant to acid (low pH) and salt stress is of industrial importance for several bioproduction processes. To identify new candidate genes having potential roles in low-pH tolerance, we screened an expression genomic DNA library of a multiple-stress-tolerant yeast, Issatchenkia orientalis (Pichia kudriavzevii), for clones that allowed Saccharomyces cerevisiae cells to grow under highly acidic conditions (pH 2.0). A genomic DNA clone containing two putative open reading frames was obtained, of which the putative protein-coding gene comprising 1629 bp was retransformed into the host. This transformant grew significantly at pH 2.0, and at pH 2.5 in the presence of 7.5% Na2SO4. The predicted amino acid sequence of this new gene, named I. orientalis GAS1 (IoGAS1), was 60% identical to the S. cerevisiae Gas1 protein, a glycosylphosphatidylinositol-anchored protein essential for maintaining cell wall integrity, and 58-59% identical to Candida albicans Phr1 and Phr2, pH-responsive proteins implicated in cell wall assembly and virulence. Northern hybridization analyses indicated that, as for the C. albicans homologs, IoGAS1 expression was pH-dependent, with expression increasing with decreasing pH (from 4.0 to 2.0) of the medium. These results suggest that IoGAS1 represents a novel pH-regulated system required for the adaptation of I. orientalis to environments of diverse pH. Heterologous expression of IoGAS1 complemented the growth and morphological defects of a S. cerevisiae gas1Δ mutant, demonstrating that IoGAS1 and the corresponding S. cerevisiae gene play similar roles in cell wall biosynthesis. Site-directed mutagenesis experiments revealed that two conserved glutamate residues (E161 and E262) in the IoGas1 protein play a crucial role in yeast morphogenesis and tolerance to low pH and salt stress. Furthermore, overexpression of IoGAS1 in S. cerevisiae remarkably improved the ethanol fermentation ability at pH 2.5, and at pH 2.0 in the presence of

  2. Anchoring visions in organizations

    Simonsen, Jesper

    1999-01-01

    This paper introduces the term 'anchoring' within systems development: Visions, developed through early systems design within an organization, need to be deeply rooted in the organization. A vision's rationale needs to be understood by those who decide if the vision should be implemented as well as...... anchoring is considered to be a main activity, is outlined. The task of anchoring visions is described, and techniques and activities are suggested with respect to those actors that have to act on the visions and the recommendations from a design proposal. The paper concludes that obtaining appropriate...

  3. Susceptibility to anchoring effects

    Todd McElroy; Keith Dowd

    2007-01-01

    Previous research on anchoring has shown this heuristic to be a very robust psychological phenomenon ubiquitous across many domains of human judgment and decision-making. Despite the prevalence of anchoring effects, researchers have only recently begun to investigate the underlying factors responsible for how and in what ways a person is susceptible to them. This paper examines how one such factor, the Big-Five personality trait of openness-to-experience, influences the effect of previously p...

  4. Gpr161 anchoring of PKA consolidates GPCR and cAMP signaling.

    Bachmann, Verena A; Mayrhofer, Johanna E; Ilouz, Ronit; Tschaikner, Philipp; Raffeiner, Philipp; Röck, Ruth; Courcelles, Mathieu; Apelt, Federico; Lu, Tsan-Wen; Baillie, George S; Thibault, Pierre; Aanstad, Pia; Stelzl, Ulrich; Taylor, Susan S; Stefan, Eduard

    2016-07-12

    Scaffolding proteins organize the information flow from activated G protein-coupled receptors (GPCRs) to intracellular effector cascades both spatially and temporally. By this means, signaling scaffolds, such as A-kinase anchoring proteins (AKAPs), compartmentalize kinase activity and ensure substrate selectivity. Using a phosphoproteomics approach we identified a physical and functional connection between protein kinase A (PKA) and Gpr161 (an orphan GPCR) signaling. We show that Gpr161 functions as a selective high-affinity AKAP for type I PKA regulatory subunits (RI). Using cell-based reporters to map protein-protein interactions, we discovered that RI binds directly and selectively to a hydrophobic protein-protein interaction interface in the cytoplasmic carboxyl-terminal tail of Gpr161. Furthermore, our data demonstrate that a binary complex between Gpr161 and RI promotes the compartmentalization of Gpr161 to the plasma membrane. Moreover, we show that Gpr161, functioning as an AKAP, recruits PKA RI to primary cilia in zebrafish embryos. We also show that Gpr161 is a target of PKA phosphorylation, and that mutation of the PKA phosphorylation site affects ciliary receptor localization. Thus, we propose that Gpr161 is itself an AKAP and that the cAMP-sensing Gpr161:PKA complex acts as cilium-compartmentalized signalosome, a concept that now needs to be considered in the analyzing, interpreting, and pharmaceutical targeting of PKA-associated functions. PMID:27357676

  5. An internally positioned signal can direct attachment of a glycophospholipid membrane anchor

    1991-01-01

    All known glycophosphatidylinositol (GPI)-anchored membrane proteins contain a COOH-terminal hydrophobic domain necessary for signalling anchor attachment. To examine the requirement that this signal be at the COOH terminus of the protein, we constructed a chimeric protein, DAFhGH, in which human growth hormone (hGH) was fused to the COOH terminus of decay accelerating factor (DAF) (a GPI-anchored protein), thereby placing the GPI signal in the middle of the chimeric protein. We show that the...

  6. Anchoring of FRET Sensors-A Requirement for Spatiotemporal Resolution.

    Ivanova, Elena V; Figueroa, Ricardo A; Gatsinzi, Tom; Hallberg, Einar; Iverfeldt, Kerstin

    2016-01-01

    FRET biosensors have become a routine tool for investigating mechanisms and components of cell signaling. Strategies for improving them for particular applications are continuously sought. One important aspect to consider when designing FRET probes is the dynamic distribution and propagation of signals within living cells. We have addressed this issue by directly comparing an anchored (taFS) to a non-anchored (naFS) cleavable FRET sensor. We chose a microtubule-associated protein tau as an anchor, as microtubules are abundant throughout the cytosol of cells. We show that tau-anchored FRET sensors are concentrated at the cytoskeleton and enriched in the neurite-like processes of cells, providing high intensity of the total signal. In addition, anchoring limits the diffusion of the sensor, enabling spatiotemporally resolved monitoring of subcellular variations in enzyme activity. Thus, anchoring is an important aspect to consider when designing FRET sensors for deeper understanding of cell signaling. PMID:27196902

  7. The calmodulin-like proteins AtCML4 and AtCML5 are single-pass membrane proteins targeted to the endomembrane system by an N-terminal signal anchor sequence.

    Ruge, Henning; Flosdorff, Sandra; Ebersberger, Ingo; Chigri, Fatima; Vothknecht, Ute C

    2016-06-01

    Calmodulins (CaMs) are important mediators of Ca(2+) signals that are found ubiquitously in all eukaryotic organisms. Plants contain a unique family of calmodulin-like proteins (CMLs) that exhibit greater sequence variance compared to canonical CaMs. The Arabidopsis thaliana proteins AtCML4 and AtCML5 are members of CML subfamily VII and possess a CaM domain comprising the characteristic double pair of EF-hands, but they are distinguished from other members of this subfamily and from canonical CaMs by an N-terminal extension of their amino acid sequence. Transient expression of yellow fluorescent protein-tagged AtCML4 and AtCML5 under a 35S-promoter in Nicotiana benthamiana leaf cells revealed a spherical fluorescence pattern. This pattern was confirmed by transient expression in Arabidopsis protoplasts under the native promoter. Co-localization analyses with various endomembrane marker proteins suggest that AtCML4 and AtCML5 are localized to vesicular structures in the interphase between Golgi and the endosomal system. Further studies revealed AtCML5 to be a single-pass membrane protein that is targeted into the endomembrane system by an N-terminal signal anchor sequence. Self-assembly green fluorescent protein and protease protection assays support a topology with the CaM domain exposed to the cytosolic surface and not the lumen of the vesicles, indicating that AtCML5 could sense Ca(2+) signals in the cytosol. Phylogenetic analysis suggests that AtCML4 and AtCML5 are closely related paralogues originating from a duplication event within the Brassicaceae family. CML4/5-like proteins seem to be universally present in eudicots but are absent in some monocots. Together these results show that CML4/5-like proteins represent a flowering plant-specific subfamily of CMLs with a potential function in vesicle transport within the plant endomembrane system. PMID:27029353

  8. Anchoring the Deficit of the Anchor Deficit: Dyslexia or Attention?

    Willburger, Edith; Landerl, Karin

    2010-01-01

    In the anchoring deficit hypothesis of dyslexia ("Trends Cogn. Sci.", 2007; 11: 458-465), it is proposed that perceptual problems arise from the lack of forming a perceptual anchor for repeatedly presented stimuli. A study designed to explicitly test the specificity of the anchoring deficit for dyslexia is presented. Four groups, representing all…

  9. A glycophospholipid membrane anchor acts as an apical targeting signal in polarized epithelial cells

    1989-01-01

    Glycosyl-phosphatidylinositol- (GPI) anchored proteins contain a large extracellular protein domain that is linked to the membrane via a glycosylated form of phosphatidylinositol. We recently reported the polarized apical distribution of all endogenous GPI-anchored proteins in the MDCK cell line (Lisanti, M. P., M. Sargiacomo, L. Graeve, A. R. Saltiel, and E. Rodriguez-Boulan. 1988. Proc. Natl. Acad. Sci. USA. 85:9557-9561). To study the role of this mechanism of membrane anchoring in targeti...

  10. The Anchoring Concept

    Simonsen, Jesper

    1998-01-01

    This paper introduces the term 'anchoring' within systems development: Visions, developed through early systems design within an organization, need to be deeply rooted in the organization. A vision's rationale needs to be understood by those who decide if the vision should be implemented as well as...... by those involved in the actual implementation. A model depicting a recent trend within systems development is presented: Organizations rely on purchasing generic software products and/or software development outsourced to external contractors. A contemporary method for participatory design, where...

  11. Release of Glycoprotein (GP1 from the Tegumental Surface of Taenia solium by Phospholipase C from Clostridium perfringens Suggests a Novel Protein-Anchor to Membranes

    Abraham Landa

    2010-01-01

    Full Text Available In order to explore how molecules are linked to the membrane surface in larval Taenia solium, whole cysticerci were incubated in the presence of phospholipase C from Clostridium perfringens (PLC. Released material was collected and analyzed in polyacrylamide gels with sodium dodecyl sulfate. Two major bands with apparent molecular weights of 180 and 43 kDa were observed. Western blot of released material and localization assays in cysticerci tissue sections using antibodies against five known surface glycoproteins of T. solium cysticerci indicated that only one, previously called GP1, was released. Similar localization studies using the lectins wheat-germ-agglutinin and Concanavalin A showed that N-acetyl-D-glucosamine, N-acetylneuraminic, sialic acid, αmethyl-D-mannoside, D-manose/glucose, and N-acetyl-D-glucosamine residues are abundantly present on the surface. On the other hand, we find that treatment with PLC releases molecules from the surface; they do not reveal Cross Reacting Determinant (CRD, suggesting a novel anchor to the membrane for the glycoprotein GP1.

  12. The membrane bound LRR lipoprotein Slr, and the cell wall-anchored M1 protein from Streptococcus pyogenes both interact with type I collagen.

    Marta Bober

    Full Text Available Streptococcus pyogenes is an important human pathogen and surface structures allow it to adhere to, colonize and invade the human host. Proteins containing leucine rich repeats (LRR have been identified in mammals, viruses, archaea and several bacterial species. The LRRs are often involved in protein-protein interaction, are typically 20-30 amino acids long and the defining feature of the LRR motif is an 11-residue sequence LxxLxLxxNxL (x being any amino acid. The streptococcal leucine rich (Slr protein is a hypothetical lipoprotein that has been shown to be involved in virulence, but at present no ligands for Slr have been identified. We could establish that Slr is a membrane attached horseshoe shaped lipoprotein by homology modeling, signal peptidase II inhibition, electron microscopy (of bacteria and purified protein and immunoblotting. Based on our previous knowledge of LRR proteins we hypothesized that Slr could mediate binding to collagen. We could show by surface plasmon resonance that recombinant Slr and purified M1 protein bind with high affinity to collagen I. Isogenic slr mutant strain (MB1 and emm1 mutant strain (MC25 had reduced binding to collagen type I as shown by slot blot and surface plasmon resonance. Electron microscopy using gold labeled Slr showed multiple binding sites to collagen I, both to the monomeric and the fibrillar structure, and most binding occurred in the overlap region of the collagen I fibril. In conclusion, we show that Slr is an abundant membrane bound lipoprotein that is co-expressed on the surface with M1, and that both these proteins are involved in recruiting collagen type I to the bacterial surface. This underlines the importance of S. pyogenes interaction with extracellular matrix molecules, especially since both Slr and M1 have been shown to be virulence factors.

  13. Bone Anchored Hearing Treatment Procedure

    Full Text Available ... to allow removal of some of the soft tissue where the screwlike fixture will be anchored. The ... series of slides demonstrates the removal of the tissue beneath the skin flap. You can see in ...

  14. Bone Anchored Hearing Treatment Procedure

    Full Text Available ... skin to allow removal of some of the soft tissue where the screwlike fixture will be anchored. The ... screen, acting as a hinge. Beneath that the soft tissue consisting of fat and some muscle is removed. ...

  15. Bone Anchored Hearing Treatment Procedure

    Full Text Available ... of the bone oscillator is superficial on the skin, the transference of that energy is less efficient, ... slides, is to mark the location in the skin where the screwlike fixture will be anchored. Subsequent ...

  16. Bone Anchored Hearing Treatment Procedure

    Full Text Available ... The Baha system consists of a bone-anchored titanium fixture that is implanted in the mastoid bone, ... the -- great. I'm going to grab the titanium implant. And what I want you to notice ...

  17. Membrane curvature enables N-Ras lipid anchor sorting to liquid-ordered membrane phases

    Larsen, Jannik Bruun; Jensen, Martin Borch; Bhatia, Vikram Kjøller;

    2015-01-01

    Trafficking and sorting of membrane-anchored Ras GTPases are regulated by partitioning between distinct membrane domains. Here, in vitro experiments and microscopic molecular theory reveal membrane curvature as a new modulator of N-Ras lipid anchor and palmitoyl chain partitioning. Membrane...... curvature was essential for enrichment in raft-like liquid-ordered phases; enrichment was driven by relief of lateral pressure upon anchor insertion and most likely affects the localization of lipidated proteins in general....

  18. Nudel Contributes to Microtubule Anchoring at the Mother Centriole and Is Involved in Both Dynein-dependent and -independent Centrosomal Protein Assembly

    Guo, Jing; Yang, Zhenye; Song, Wei; Chen, Qi; Wang, Fubin; Zhang, Qiangge; Zhu, Xueliang

    2006-01-01

    The centrosome is the major microtubule-organizing center in animal cells. Although the cytoplasmic dynein regulator Nudel interacts with centrosomes, its role herein remains unclear. Here, we show that in Cos7 cells Nudel is a mother centriole protein with rapid turnover independent of dynein activity. During centriole duplication, Nudel targets to the new mother centriole later than ninein but earlier than dynactin. Its centrosome localization requires a C-terminal region that is essential ...

  19. Poly(ethylene glycol) anchored to poly(dopamine) layers: highly versatile and simple way of suppressing protein adsorption and cell adhesion

    Pop-Georgievski, Ognen; Popelka, Štěpán; Proks, Vladimír; Houska, Milan; Jaroš, J.; Hampl, A.; Rypáček, František

    Praha : Ústav makromolekulární chemie AV ČR, 2010. L_21. ISBN 978-80-85009-62-0. [Workshop "Career in Polymers" /2./. 23.07.2010-24.07.2010, Praha] R&D Projects: GA AV ČR KJB400500904; GA MŠk 1M0538 EU Projects: European Commission(XE) 500283 - EXPERTISSUES Institutional research plan: CEZ:AV0Z40500505 Keywords : poly(ethylene glycol) * protein adsorption * biomedical applications Subject RIV: CD - Macromolecular Chemistry

  20. Anchoring a Leviathan: How the Nuclear Membrane Tethers the Genome

    Rafal eCzapiewski

    2016-05-01

    Full Text Available It is well established that the nuclear envelope has many distinct direct connections to chromatin that contribute to genome organization. The functional consequences of genome organization on gene regulation are less clear. Even less understood is how interactions of lamins and nuclear envelope transmembrane proteins (NETs with chromatin can produce anchoring tethers that can withstand the physical forces of and on the genome. Chromosomes are the largest molecules in the cell, making megadalton protein structures like the nuclear pore complexes and ribosomes seem small by comparison. Thus to withstand strong forces from chromosome dynamics an anchoring tether is likely to be much more complex than a single protein-protein or protein-DNA interaction. Here we will briefly review known NE-genome interactions that likely contribute to spatial genome organization, postulate in the context of experimental data how these anchoring tethers contribute to gene regulation, and posit several hypotheses for the physical nature of these tethers that need to be investigated experimentally. Significantly, disruption of these anchoring tethers and the subsequent consequences for gene regulation could explain how mutations in nuclear envelope proteins cause diseases ranging from muscular dystrophy to lipodystrophy to premature ageing progeroid syndromes. The two favored hypotheses for nuclear envelope protein involvement in disease are 1 weakening nuclear and cellular mechanical stability, and 2 disrupting genome organization and gene regulation. Considerable experimental support has been obtained for both. The integration of both mechanical and gene expression defects in the disruption of anchoring tethers could provide a unifying hypothesis consistent with both.

  1. Permanent ground anchors: Nicholson design criteria

    Nicholson, P. J.; Uranowski, D. D.; Wycliffe-Jones, P. T.

    1982-09-01

    The methods used by Nicholson Construction Company in the design of permanent ground anchors specifically as related to retaining walls are discussed. Basic soil parameters, design concepts, drilling and grouting methods for ground anchors are discussed. Particular emphasis is placed on anchors founded in soil rather than rock formations. Also, soil properties necessary for the proper design of anchored retaining walls are detailed. The second chapter of the report is devoted to a general discussion of retaining wall and anchor design. In addition, a design example of an anchored retaining wall is presented in a step by step manner.

  2. Type II PKAs are anchored to mature insulin secretory granules in INS-1 β-cells and required for cAMP-dependent potentiation of exocytosis.

    Villalpando, Sabrina; Cazevieille, Chantal; Fernandez, Anne; Lamb, Ned J; Hani, El-Habib

    2016-06-01

    Specificity of the cAMP-dependent protein kinase (PKA) pathway relies on an extremely sophisticated compartmentalization mechanism of the kinase within a given cell, based on high-affinity binding of PKA tetramer pools to different A-Kinase Anchoring Proteins (AKAPs). We and others have previously shown that AKAPs-dependent PKA subcellular targeting is a requisite for optimal cAMP-dependent potentiation of insulin exocytosis. We thus hypothesized that a PKA pool may directly anchor to the secretory compartment to potentiate insulin exocytosis. Here, using immunofluorescence analyses combined to subcellular fractionations and purification of insulin secretory granules (ISGs), we identified discrete subpools of type II PKAs, RIIα and RIIβ PKAs, along with the catalytic subunit, physically associated with ISGs within pancreatic insulin-secreting β-cells. Ultrastructural analysis of native rodent β-cells confirmed in vivo the occurrence of PKA on dense-core ISGs. Isoform-selective disruption of binding of PKAs to AKAPs reinforced the requirement of type II PKA isoforms for cAMP potentiation of insulin exocytosis. This granular localization of PKA was of critical importance since siRNA-mediated depletion of either RIIα or RIIβ PKAs resulted in a significant reduction of cAMP-dependent potentiation of insulin release. The present work provides evidence for a previously unrecognized pool of type II PKAs physically anchored to the β-cell ISGs compartment and supports a non-redundant function for type II PKAs during cAMP potentiation of exocytosis. PMID:26898328

  3. Structural models of the membrane anchors of envelope glycoproteins E1 and E2 from pestiviruses

    The membrane anchors of viral envelope proteins play essential roles in cell entry. Recent crystal structures of the ectodomain of envelope protein E2 from a pestivirus suggest that E2 belongs to a novel structural class of membrane fusion machinery. Based on geometric constraints from the E2 structures, we generated atomic models of the E1 and E2 membrane anchors using computational approaches. The E1 anchor contains two amphipathic perimembrane helices and one transmembrane helix; the E2 anchor contains a short helical hairpin stabilized in the membrane by an arginine residue, similar to flaviviruses. A pair of histidine residues in the E2 ectodomain may participate in pH sensing. The proposed atomic models point to Cys987 in E2 as the site of disulfide bond linkage with E1 to form E1–E2 heterodimers. The membrane anchor models provide structural constraints for the disulfide bonding pattern and overall backbone conformation of the E1 ectodomain. - Highlights: • Structures of pestivirus E2 proteins impose constraints on E1, E2 membrane anchors. • Atomic models of the E1 and E2 membrane anchors were generated in silico. • A “snorkeling” arginine completes the short helical hairpin in the E2 membrane anchor. • Roles in pH sensing and E1–E2 disulfide bond formation are proposed for E1 residues. • Implications for E1 ectodomain structure and disulfide bonding pattern are discussed

  4. Structural models of the membrane anchors of envelope glycoproteins E1 and E2 from pestiviruses

    Wang, Jimin, E-mail: jimin.wang@yale.edu; Li, Yue; Modis, Yorgo, E-mail: yorgo.modis@yale.edu

    2014-04-15

    The membrane anchors of viral envelope proteins play essential roles in cell entry. Recent crystal structures of the ectodomain of envelope protein E2 from a pestivirus suggest that E2 belongs to a novel structural class of membrane fusion machinery. Based on geometric constraints from the E2 structures, we generated atomic models of the E1 and E2 membrane anchors using computational approaches. The E1 anchor contains two amphipathic perimembrane helices and one transmembrane helix; the E2 anchor contains a short helical hairpin stabilized in the membrane by an arginine residue, similar to flaviviruses. A pair of histidine residues in the E2 ectodomain may participate in pH sensing. The proposed atomic models point to Cys987 in E2 as the site of disulfide bond linkage with E1 to form E1–E2 heterodimers. The membrane anchor models provide structural constraints for the disulfide bonding pattern and overall backbone conformation of the E1 ectodomain. - Highlights: • Structures of pestivirus E2 proteins impose constraints on E1, E2 membrane anchors. • Atomic models of the E1 and E2 membrane anchors were generated in silico. • A “snorkeling” arginine completes the short helical hairpin in the E2 membrane anchor. • Roles in pH sensing and E1–E2 disulfide bond formation are proposed for E1 residues. • Implications for E1 ectodomain structure and disulfide bonding pattern are discussed.

  5. Endoplasmic reticulum anchored heme-oxygenase 1 faces the cytosol

    Gottlieb, Yehonatan; Truman, Marianna; Cohen, Lyora A.; Leichtmann-Bardoogo, Yael; Meyron-Holtz, Esther G.

    2012-01-01

    Heme-oxygenase 1 is an endoplasmic reticulum-anchored enzyme that breaks down heme into iron, carbon monoxide and biliverdin. Heme is a hydrophobic co-factor in many proteins, including hemoglobin. Free heme is highly cytotoxic and, therefore, both heme synthesis and breakdown are tightly regulated. During turnover of heme proteins, heme is released in the phago-lysosomal compartment or the cytosol. The subcellular location of the heme-oxygenase 1 active site has not been clarified. Using con...

  6. Inconspicuous anchoring effects generated by false information

    Chen Qu; Jun Wang; Yuejia Luo

    2008-01-01

    The impact of false information on numerical judgments was examined on young normal subjects by an event-related potential (ERP) experiment. To imitate the judgments in real world, we ensured the subjects acknowledged of the target task. The behavioral results found that both uncertain information and false information assimilated the final estimates: higher after higher anchors and lower after lower anchors; and false information caused a weaker anchoring bias than uncertain information. ERP results provided further electrophysiological evidence for the mechanism of anchoring. In the early phrase, it was an accessibility-dominated process in which two kinds of anchors elicited an N300 component related to the accessibility of anchors propositions. The knowledge relevant to targets joined the process in the late phrase, which caused a larger amplitude of late positive component (LPC) for implausible lower anchors than that for plausible higher anchors. Source analysis showed that medial frontal gyrus, whose activity was suggested to signal the need of adjustment, was more reliable to explain the LPC elicited by implausible lower anchors. Therefore, we suggest that accessibility is facilitated when the external anchor is consistent with the world knowledge, and adjustment is initiated when the external anchor is inconsistent.

  7. Anchoring in numeric judgments of visual stimuli

    Linda eLangeborg

    2016-02-01

    Full Text Available This article investigates effects of anchoring in age estimation and estimation of quantities, two tasks which to different extents are based on visual stimuli. The results are compared to anchoring in answers to classic general knowledge questions that rely on semantic knowledge. Cognitive load was manipulated to explore possible differences between domains. Effects of source credibility, manipulated by differing instructions regarding the selection of anchor values (no information regarding anchor selection, information that the anchors are randomly generated or information that the anchors are answers from an expert on anchoring were also investigated. Effects of anchoring were large for all types of judgments but were not affected by cognitive load or by source credibility in either one of the researched domains. A main effect of cognitive load on quantity estimations and main effects of source credibility in the two visually based domains indicate that the manipulations were efficient. Implications for theoretical explanations of anchoring are discussed. In particular, because anchoring did not interact with source credibility or cognitive load, the results imply that the process behind anchoring in visual tasks is predominantly automatic and unconscious.

  8. Anchoring in Numeric Judgments of Visual Stimuli.

    Langeborg, Linda; Eriksson, Mårten

    2016-01-01

    This article investigates effects of anchoring in age estimation and estimation of quantities, two tasks which to different extents are based on visual stimuli. The results are compared to anchoring in answers to classic general knowledge questions that rely on semantic knowledge. Cognitive load was manipulated to explore possible differences between domains. Effects of source credibility, manipulated by differing instructions regarding the selection of anchor values (no information regarding anchor selection, information that the anchors are randomly generated or information that the anchors are answers from an expert) on anchoring were also investigated. Effects of anchoring were large for all types of judgments but were not affected by cognitive load or by source credibility in either one of the researched domains. A main effect of cognitive load on quantity estimations and main effects of source credibility in the two visually based domains indicate that the manipulations were efficient. Implications for theoretical explanations of anchoring are discussed. In particular, because anchoring did not interact with cognitive load, the results imply that the process behind anchoring in visual tasks is predominantly automatic and unconscious. PMID:26941684

  9. Anchors and their use in offshore petroleum operations

    1984-01-01

    Holding a ship or vessel in a fixed position over specific spots is essential in drilling subsea wells. Conventional anchors and holding systems have thus been transformed into permanent mooring points capable of withstanding enormous stresses. This book specifies the parameters directly governing behavior of these new anchors, studies the mechanisms involved in anchor/seabed interaction and interprets these in the context of soil mechanics, and draws practical conclusions in the form of methods for predicting holding capacity in different types of subsea soils. Contents are: history of anchors; high-capacity anchoring systems; kinetics of anchors; holding power of anchors; anchoring site survey; choice of an anchor; and anchoring tests.

  10. Shifting Nominal Anchors: The Experience of Mexico

    Leonardo Leiderman; Nissan Liviatan; Alfredo Thorne

    1995-01-01

    In the mid-1980’s Mexico successfully brought down its high rate of inflation by using the exchange rate as nominal anchor in combination with strict fiscal discipline, tight monetary policy, and incomes policy. This paper discusses the role of exchange rate policy as nominal anchor in Mexico and develops the inflation target as the monetary framework for anchoring prices. It also describes how Mexico is applying this frame work while shifting to a more flexible exchange regime and discusses ...

  11. Capital Asset Pricing Model Adjusted for Anchoring

    Hammad, Siddiqi

    2015-01-01

    I show that adjusting CAPM for anchoring provides a unified explanation for the size, value, and momentum effects. Anchoring adjusted CAPM (ACAPM) predicts that stock splits are associated with positive abnormal returns and an increase in return volatility, whereas the reverse stock-splits are associated with negative abnormal returns and a fall in return volatility. Existing empirical evidence strongly supports these predictions. Anchoring has the effect of pushing up the equity premium, a ...

  12. PredGPI: a GPI-anchor predictor

    Casadio Rita

    2008-09-01

    Full Text Available Abstract Background Several eukaryotic proteins associated to the extracellular leaflet of the plasma membrane carry a Glycosylphosphatidylinositol (GPI anchor, which is linked to the C-terminal residue after a proteolytic cleavage occurring at the so called ω-site. Computational methods were developed to discriminate proteins that undergo this post-translational modification starting from their aminoacidic sequences. However more accurate methods are needed for a reliable annotation of whole proteomes. Results Here we present PredGPI, a prediction method that, by coupling a Hidden Markov Model (HMM and a Support Vector Machine (SVM, is able to efficiently predict both the presence of the GPI-anchor and the position of the ω-site. PredGPI is trained on a non-redundant dataset of experimentally characterized GPI-anchored proteins whose annotation was carefully checked in the literature. Conclusion PredGPI outperforms all the other previously described methods and is able to correctly replicate the results of previously published high-throughput experiments. PredGPI reaches a lower rate of false positive predictions with respect to other available methods and it is therefore a costless, rapid and accurate method for screening whole proteomes.

  13. DEX-1 and DYF-7 establish sensory dendrite length by anchoring dendritic tips during cell migration

    Heiman, Maxwell G.; Shaham, Shai

    2009-01-01

    Cells are devices whose structures delimit function. For example, in the nervous system, neuronal and glial shapes dictate paths of information flow. To understand how cells acquire their shapes, we examined the formation of a sense organ in C. elegans. Using time-lapse imaging, we found that sensory dendrites form by stationary anchoring of dendritic tips during cell-body migration. A genetic screen identified DEX-1 and DYF-7, extracellular proteins required for dendritic tip anchoring, whic...

  14. Method of fabrication of anchored nanostructure materials

    Seals, Roland D; Menchhofer, Paul A; Howe, Jane Y; Wang, Wei

    2013-11-26

    Methods for fabricating anchored nanostructure materials are described. The methods include heating a nano-catalyst under a protective atmosphere to a temperature ranging from about 450.degree. C. to about 1500.degree. C. and contacting the heated nano-catalysts with an organic vapor to affix carbon nanostructures to the nano-catalysts and form the anchored nanostructure material.

  15. Suction anchors for floating production systems

    Tjelta, T.I.; Rusas, P. [Statoil a.s. (Norway); Edvardsen, G. [HEX a.s. (Norway)

    1996-12-31

    Since the suction anchor is a recent development in its current use, this work provides a brief historical background. It is shown that the suction anchor throughout evolution today is a competitive foundation solution for many applications. Examples from these categories are included in this paper. Also several installation methods are discussed. 3 refs., 7 figs., 2 tabs.

  16. Regulation of ADAM12 cell-surface expression by protein kinase C epsilon

    Sundberg, Christina; Thodeti, Charles Kumar; Kveiborg, Marie;

    2004-01-01

    constitutively active protein. However, little is known about the regulation of ADAM12 cell-surface translocation. Here, we used human RD rhabdomyosarcoma cells, which express ADAM12 at the cell surface, in a temporal pattern. We report that protein kinase C (PKC) epsilon induces ADAM12 translocation to the cell......The ADAM (a disintegrin and metalloprotease) family consists of multidomain cell-surface proteins that have a major impact on cell behavior. These transmembrane-anchored proteins are synthesized as proforms that have (from the N terminus): a prodomain; a metalloprotease-, disintegrin......-immunoprecipitated from membrane-enriched fractions of PMA-treated cells, 3) RD cells transfected with EGFP-tagged, myristoylated PKCepsilon expressed more ADAM12 at the cell surface than did non-transfected cells, and 4) RD cells transfected with a kinase-inactive PKCepsilon mutant did not exhibit ADAM12 cell...

  17. Construction and characterization of secreted and chimeric transmembrane forms of Drosophila acetylcholinesterase: a large truncation of the C-terminal signal peptide does not eliminate glycoinositol phospholipid anchoring.

    Incardona, J P; Rosenberry, T. L.

    1996-01-01

    Despite advances in understanding the cell biology of glycoinositol phospholipid (GPI)-anchored proteins in cultured cells, the in vivo functions of GPI anchors have remained elusive. We have focused on Drosophila acetylcholinesterase (AChE) as a model GPI-anchored protein that can be manipulated in vivo with sophisticated genetic techniques. In Drosophila, AChE is found only as a GPI-anchored G2 form encoded by the Ace locus on the third chromosome. To pursue our goal of replacing wild-type ...

  18. Anchored nanostructure materials and method of fabrication

    Seals, Roland D; Menchhofer, Paul A; Howe, Jane Y; Wang, Wei

    2012-11-27

    Anchored nanostructure materials and methods for their fabrication are described. The anchored nanostructure materials may utilize nano-catalysts that include powder-based or solid-based support materials. The support material may comprise metal, such as NiAl, ceramic, a cermet, or silicon or other metalloid. Typically, nanoparticles are disposed adjacent a surface of the support material. Nanostructures may be formed as anchored to nanoparticles that are adjacent the surface of the support material by heating the nano-catalysts and then exposing the nano-catalysts to an organic vapor. The nanostructures are typically single wall or multi-wall carbon nanotubes.

  19. Mooring Anchors for Marine Renewable Energy Foundations

    Stevens, Robert F.; Rahim, Amir

    2014-01-01

    With the increasing use of offshore wind turbines, it has become necessary to explore deep-water sites for locating wind farms. Floating turbines are an ideal choice for these locations. Such turbines are anchored with mooring chains to the sea floor using suction anchors, driven piles or gravity foundations. This paper presents design methods for these types of foundations. Moored gravity foundations have been used for the much larger floating oil and gas installations. These concrete founda...

  20. Anchoring Adjusted Capital Asset Pricing Model

    Hammad, Siddiqi

    2015-01-01

    An anchoring adjusted Capital Asset Pricing Model (ACAPM) is developed in which the payoff volatilities of well-established stocks are used as starting points that are adjusted to form volatility judgments about other stocks. Anchoring heuristic implies that such adjustments are typically insufficient. ACAPM converges to CAPM with correct adjustment, so CAPM is a special case of ACAPM. The model provides a unified explanation for the size, value, and momentum effects in the stock market. A ke...

  1. FRP-to-concrete joint assemblies anchored with multiple FRP anchors: experimental investigation

    Zhang, H.; Smith, ST

    2010-01-01

    Higher strains can be developed in fibre-reinforced polymer (FRP) composites which are bonded to the surfaces of concrete members if the FRP is anchored. Anchors made from FRP (also known as FRP spike anchors but herein referred to as FRP anchors) are a promising type of anchorage as they can be applied to a variety of different shaped structural elements and they have been shown to be effective in en-hancing the strain capacity of externally bonded FRP. Limited research, however, has been co...

  2. DEX-1 and DYF-7 establish sensory dendrite length by anchoring dendritic tips during cell migration.

    Heiman, Maxwell G; Shaham, Shai

    2009-04-17

    Cells are devices whose structures delimit function. For example, in the nervous system, neuronal and glial shapes dictate paths of information flow. To understand how cells acquire their shapes, we examined the formation of a sense organ in C. elegans. Using time-lapse imaging, we found that sensory dendrites form by stationary anchoring of dendritic tips during cell-body migration. A genetic screen identified DEX-1 and DYF-7, extracellular proteins required for dendritic tip anchoring, which act cooperatively at the time and place of anchoring. DEX-1 and DYF-7 contain, respectively, zonadhesin and zona pellucida domains, and DYF-7 self-associates into multimers important for anchoring. Thus, unlike other dendrites, amphid dendritic tips are positioned by DEX-1 and DYF-7 without the need for long-range guidance cues. In sequence and function, DEX-1 and DYF-7 resemble tectorins, which anchor stereocilia in the inner ear, suggesting that a sensory dendrite anchor may have evolved into part of a mechanosensor. PMID:19344940

  3. Anchors as Semantic Primes in Value Construction: An EEG Study of the Anchoring Effect.

    Qingguo Ma

    Full Text Available Previous research regarding anchoring effects has demonstrated that human judgments are often assimilated to irrelevant information. Studies have demonstrated that anchors influence the economic valuation of various products and experiences; however, the cognitive explanations of this effect remain controversial, and its neural mechanisms have rarely been explored. In the current study, we conducted an electroencephalography (EEG experiment to investigate the anchoring effect on willingness to accept (WTA for an aversive hedonic experience and the role of anchors in this judgment heuristic. The behavioral results demonstrated that random numbers affect participants' WTA for listening to pieces of noise. The participants asked for higher pay after comparing their WTA with higher numbers. The EEG results indicated that anchors also influenced the neural underpinnings of the valuation process. Specifically, when a higher anchor number was drawn, larger P2 and late positive potential amplitudes were elicited, reflecting the anticipation of more intensive pain from the subsequent noise. Moreover, higher anchors induced a stronger theta band power increase compared with lower anchors when subjects listened to the noises, indicating that the participants felt more unpleasant during the actual experience of the noise. The levels of unpleasantness during both anticipation and experience were consistent with the semantic information implied by the anchors. Therefore, these data suggest that a semantic priming process underlies the anchoring effect in WTA. This study provides proof for the robustness of the anchoring effect and neural evidence of the semantic priming model. Our findings indicate that activated contextual information, even seemingly irrelevant, can be embedded in the construction of economic value in the brain.

  4. Applying a Targeted Label-free Approach using LC-MS AMT Tags to Evaluate Changes in Protein Phosphorylation Following Phosphatase Inhibition

    Yang, Feng; Jaitly, Navdeep; Jayachandran, Hemalatha; Lou, Quanzhou; Monroe, Matthew E.; Du, Xiuxia; Gritsenko, Marina A.; Zhang, Rui; Anderson, David J.; Purvine, Samuel O.; Adkins, Joshua N.; Moore, Ronald J.; Mottaz, Heather M.; Ding, Shi-Jian; Lipton, Mary S.; Camp, David G.; Udseth, Harold R.; Smith, Richard D.; Rossie, Sandra S.

    2007-10-12

    To identify phosphoproteins regulated by the phosphoprotein phosphatase (PPP) family of S/T phosphatases, we performed a large-scale characterization of changes in protein phosphorylation on extracts from HeLa cells treated with or without calyculin A, a potent PPP enzyme inhibitor. A label-free comparative Phosphoproteomics approach using immobilized metal ion affinity chromatography and targeted tandem mass spectrometry was employed to discover and identify signatures based upon distinctive changes in abundance. Overall, 232 proteins were identified as either direct or indirect targets for PPP enzyme regulation. Most of the present identifications represent novel PPP enzyme targets at the level of both phosphorylation site and protein. These include phosphorylation sites within signaling proteins such as p120 Catenin, A Kinase Anchoring Protein 8, JunB, and Type II Phosphatidyl Inositol 4 Kinase. These data can be used to define underlying signaling pathways and events regulated by the PPP family of S/T phosphatases.

  5. Construction of a highly active xylanase displaying oleaginous yeast: comparison of anchoring systems.

    Sophie Duquesne

    Full Text Available Three Yarrowia lipolytica cell wall proteins (YlPir, YlCWP1 and YlCBM were evaluated for their ability to display the xylanase TxXYN from Thermobacillus xylanilyticus on the cell surface of Y. lipolytica. The fusion proteins were produced in Y. lipolytica JMY1212, a strain engineered for mono-copy chromosomal insertion, and enabling accurate comparison of anchoring systems. The construction using YlPir enabled cell bound xylanase activity to be maximised (71.6 U/g. Although 48% of the activity was released in the supernatant, probably due to proteolysis at the fusion zone, this system is three times more efficient for the anchoring of TxXYN than the YlCWP1 system formerly developed for Y. lipolytica. As far as we know it represents the best displayed xylanase activity ever published. It could be an attractive alternative anchoring system to display enzymes in Y. lipolytica.

  6. Monitoring ground anchor using non-destructive ground anchor integrity test (NDT-GRANIT)

    Monitoring at ground anchor commonly uses a pull out test method, therefor we developing a non-destructive ground anchor integrity testing (NDT-GRANIT). NDT-GRANIT using the principle of seismic waves that have been modified into form of sweep signal, the signal will be demodulated, filtered, and Fourier transformation (inverse discrete Fourier transform) so the data can be interpreted reflected wave from the ground anchor. The method was applied to determine whether the ground anchor still gripped in the subsurface by looking the attenuation of the wave generated sources. From the result we can see that ground anchor does not grip. To validate the results of the comparison method of measurement used pile integrity test

  7. ANCHORING EFFECT ANALYSIS OF TENSIONED BOLTS

    夏建中

    1997-01-01

    The paper analyses quantitatively the anchoring effect of tensioned bolts on surrounding rock strength, and defines two concepts: one is the surrounding rock strength increased amount △τ13 and the other is the strength influence factor k. The anchoring effect of tensioned bolts is considered to increase a strength increased amount △τ13 where △τ13 is the product k and tensioned load p, i. e. △τ13= kp, where k is a function of two variables x and y. The distributive properties both △τ13 and k are also discussed in the paper, obtaining some useful results for designing bolting support parameters.

  8. Monogenean anchor morphometry: systematic value, phylogenetic signal, and evolution.

    Khang, Tsung Fei; Soo, Oi Yoon Michelle; Tan, Wooi Boon; Lim, Lee Hong Susan

    2016-01-01

    Background. Anchors are one of the important attachment appendages for monogenean parasites. Common descent and evolutionary processes have left their mark on anchor morphometry, in the form of patterns of shape and size variation useful for systematic and evolutionary studies. When combined with morphological and molecular data, analysis of anchor morphometry can potentially answer a wide range of biological questions. Materials and Methods. We used data from anchor morphometry, body size and morphology of 13 Ligophorus (Monogenea: Ancyrocephalidae) species infecting two marine mugilid (Teleostei: Mugilidae) fish hosts: Moolgarda buchanani (Bleeker) and Liza subviridis (Valenciennes) from Malaysia. Anchor shape and size data (n = 530) were generated using methods of geometric morphometrics. We used 28S rRNA, 18S rRNA, and ITS1 sequence data to infer a maximum likelihood phylogeny. We discriminated species using principal component and cluster analysis of shape data. Adams's K mult was used to detect phylogenetic signal in anchor shape. Phylogeny-correlated size and shape changes were investigated using continuous character mapping and directional statistics, respectively. We assessed morphological constraints in anchor morphometry using phylogenetic regression of anchor shape against body size and anchor size. Anchor morphological integration was studied using partial least squares method. The association between copulatory organ morphology and anchor shape and size in phylomorphospace was used to test the Rohde-Hobbs hypothesis. We created monogeneaGM, a new R package that integrates analyses of monogenean anchor geometric morphometric data with morphological and phylogenetic data. Results. We discriminated 12 of the 13 Ligophorus species using anchor shape data. Significant phylogenetic signal was detected in anchor shape. Thus, we discovered new morphological characters based on anchor shaft shape, the length between the inner root point and the outer root

  9. Stone anchors from Minicoy Island, Lakshadweep, India

    Tripati, S.

    at Androth, Kadmat, Kavaratti and Minicoy. In addition to these finds, in recent past an Indo-Arabian type of stone anchor has come to lime light at the Jama Mosque of Funhilol of Minicoy. In this paper an attempt has been made to describe the Indo...

  10. Anchor Stress Checking of Security Injection Tank

    2011-01-01

    The intention of the calculating is to check the anchor stresses of the security injection tank to know whether the stress is satisfied the code requirements on the basis of all the reaction forces gained in the static, seismic and thermal stress results.

  11. Predicting Anchor Links between Heterogeneous Social Networks

    Sajadmanesh, Sina; Khodadadi, Ali

    2016-01-01

    People usually get involved in multiple social networks to enjoy new services or to fulfill their needs. Many new social networks try to attract users of other existing networks to increase the number of their users. Once a user (called source user) of a social network (called source network) joins a new social network (called target network), a new inter-network link (called anchor link) is formed between the source and target networks. In this paper, we concentrated on predicting the formation of such anchor links between heterogeneous social networks. Unlike conventional link prediction problems in which the formation of a link between two existing users within a single network is predicted, in anchor link prediction, the target user is missing and will be added to the target network once the anchor link is created. To solve this problem, we use meta-paths as a powerful tool for utilizing heterogeneous information in both the source and target networks. To this end, we propose an effective general meta-pat...

  12. Influence of Anchoring on Burial Depth of Submarine Pipelines.

    Zhuang, Yuan; Li, Yang; Su, Wei

    2016-01-01

    Since the beginning of the twenty-first century, there has been widespread construction of submarine oil-gas transmission pipelines due to an increase in offshore oil exploration. Vessel anchoring operations are causing more damage to submarine pipelines due to shipping transportation also increasing. Therefore, it is essential that the influence of anchoring on the required burial depth of submarine pipelines is determined. In this paper, mathematical models for ordinary anchoring and emergency anchoring have been established to derive an anchor impact energy equation for each condition. The required effective burial depth for submarine pipelines has then been calculated via an energy absorption equation for the protection layer covering the submarine pipelines. Finally, the results of the model calculation have been verified by accident case analysis, and the impact of the anchoring height, anchoring water depth and the anchor weight on the required burial depth of submarine pipelines has been further analyzed. PMID:27166952

  13. Influence of Anchoring on Burial Depth of Submarine Pipelines

    Zhuang, Yuan; Li, Yang; Su, Wei

    2016-01-01

    Since the beginning of the twenty-first century, there has been widespread construction of submarine oil-gas transmission pipelines due to an increase in offshore oil exploration. Vessel anchoring operations are causing more damage to submarine pipelines due to shipping transportation also increasing. Therefore, it is essential that the influence of anchoring on the required burial depth of submarine pipelines is determined. In this paper, mathematical models for ordinary anchoring and emergency anchoring have been established to derive an anchor impact energy equation for each condition. The required effective burial depth for submarine pipelines has then been calculated via an energy absorption equation for the protection layer covering the submarine pipelines. Finally, the results of the model calculation have been verified by accident case analysis, and the impact of the anchoring height, anchoring water depth and the anchor weight on the required burial depth of submarine pipelines has been further analyzed. PMID:27166952

  14. Ice nucleation protein as a bacterial surface display protein

    Sarhan Mohammed A.A.

    2011-01-01

    Surface display technology can be defined as that phenotype (protein or peptide) which is linked to a genotype (DNA or RNA) through an appropriate anchoring motif. A bacterial surface display system is based on expressing recombinant proteins fused to sorting signals (anchoring motifs) that direct their incorporation on the cell surface.

  15. Maleimide-functionalized lipids that anchor polypeptides to lipid bilayers and membranes.

    Elliott, J T; Prestwich, G D

    2000-01-01

    Two maleimide-containing diacylglycerol derivatives were synthesized to permit the anchoring of short peptides and longer polypeptides to phospholipid bilayers and membranes. The maleimide was introduced at the site normally occupied by a phospholipid headgroup. The first lipid, the dipalmitoyl ester of 1-maleimido-2,3-propanediol, was developed as a membrane anchor for extracellular domains of transmembrane proteins. The second anchoring lipid, in which the 3-position contained a 6-aminohexanoate, was designed for convenient modification with amine-reactive reporter groups. Specifically, the NBD fluorophore, 7-nitrobenzo-2-oxa-1, 3-diazole-aminohexanoic-N-hydroxysuccinimide ester, was attached to give an fluorescent anchoring reagent. Next, these reagents were applied to the anchoring of a C-terminally cysteamine-modified 8 kDa polypeptide that comprises the extracellular N-terminal domain of the human thrombin receptor, a transmembrane protease-activated receptor (PAR-1). Gel filtration and fluorescence analysis showed that the fluorescent lipopolypeptide spontaneously inserted into preformed phospholipid vesicles, but it did not insert into whole cell membranes. In contrast, the dipalmitoyl derivative could only be reconstituted into artificial membranes by mixing the lipopolypeptide and phospholipid before vesicle formation. These results suggest that biophysical interactions governing the lipopolypeptide insertion into artificial and cellular membranes may differ. The thiol-reactive lipidating reagents should be valuable materials for studying the structure and function of peptides and polypeptides at phospholipid bilayer surfaces. PMID:11087332

  16. Stone anchors of India: Findings, classification and significance.

    Tripati, S.

    Honolulu, Hawaii, May 12-16, 2014, Vol. 2: 973-986 973 Stone anchors of India: Findings, Classification and Significance Sila Tripati CSIR-National Institute of Oceanography, Dona Paula, Goa 403 004, India Email: sila@nio.org Abstract Various.... The making of anchors from the hero stones indicates their alternative use (Sila Tripati and Patnaik, 2008). Classification Stone anchors are classified according to character, shape and region. In general stone anchors have been classified...

  17. Susceptibility to anchoring effects: How openness-to-experience influences responses to anchoring cues

    Todd McElroy; Keith Dowd

    2007-01-01

    Previous research on anchoring has shown this heuristic to be a very robust psychological phenomenon ubiquitous across many domains of human judgment and decision-making. Despite the prevalence of anchoring effects, researchers have only recently begun to investigate the underlying factors responsible for how and in what ways a person is susceptible to them. This paper examines how one such factor, the Big-Five personality trait of openness-to-experience, influences the effect of previously p...

  18. Monolayer Assemblies of a De Novo Designed 4-alpha-Helix Bundle Carboprotein and Its Sulfur Anchor Fragment on Au(111) Surfaces Addressed by Voltammetry and In Situ Scanning Tunneling Microscopy

    Brask, Jesper; Wackerbarth, Hainer; Jensen, Knud J.;

    2003-01-01

    scheme for the synthesis of a new 4-R-helix bundle carboprotein built on a galactopyranoside derivative with a thiol anchor aglycon suitable for surface immobilization on gold. The carboprotein with thiol anchor in monomeric and dimeric (disulfide) form, the thiol anchor alone, and a sulfur-free 4-R......-helix bundle carboprotein without thiol anchor have been prepared and investigated for comparison. Cyclic and differential pulse voltammetry (DPV) of the proteins show desorption peaks around -750 mV (SCE), whereas the thiol anchor desorption peak is at -685 mV. The peaks are by far the highest for thiol...... monomeric 4-R-helix bundle carboprotein and the thiol anchor. This pattern is supported by capacitance data. The DPV and capacitance data for the thiolated 4-R-helix bundle carboproteins and the thiol anchor hold a strong Faradaic reductive desorption component as supported by X-ray photoelectron...

  19. Students' Anchoring Predisposition: An Illustration from Spring Training Baseball

    Mohrweis, Lawrence C.

    2014-01-01

    The anchoring tendency results when decision makers anchor on initial values and then make final assessments that are adjusted insufficiently away from the initial values. The professional literature recognizes that auditors often risk falling into the judgment trap of anchoring and adjusting (Ranzilla et al., 2011). Students may also be unaware…

  20. 33 CFR 401.14 - Anchor marking buoys.

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Anchor marking buoys. 401.14 Section 401.14 Navigation and Navigable Waters SAINT LAWRENCE SEAWAY DEVELOPMENT CORPORATION, DEPARTMENT OF TRANSPORTATION SEAWAY REGULATIONS AND RULES Regulations Condition of Vessels § 401.14 Anchor marking buoys. A highly visible anchor marking...

  1. Lentiviral Engineered Fibroblasts Expressing Codon Optimized COL7A1 Restore Anchoring Fibrils in RDEB

    Georgiadis, Christos,; Syed, Farhatullah; Petrova, Anastasia; Abdul-Wahab, Alya; Lwin, Su M; Farzaneh, Farzin; Chan, Lucas; Ghani, Sumera; Fleck, Roland A.; Glover, Leanne; McMillan, James R.; Chen, Mei; Adrian J Thrasher; McGrath, John A; Di, Wei-Li

    2015-01-01

    Cells therapies, engineered to secrete replacement proteins, are being developed to ameliorate otherwise debilitating diseases. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects of type VII collagen (C7), a protein essential for anchoring fibril formation at the dermal-epidermal junction (DEJ). Whilst allogeneic fibroblasts injected directly into the dermis can mediate transient disease modulation, autologous gene-modified fibroblasts should evade immunological rejection ...

  2. Removal of the glycosylphosphatidylinositol anchor from PrP(Sc) by cathepsin D does not reduce prion infectivity.

    Lewis, Patrick A; Properzi, Francesca; Prodromidou, Kanella; Clarke, Anthony R; Collinge, John; Jackson, Graham S

    2006-04-15

    According to the protein-only hypothesis of prion propagation, prions are composed principally of PrP(Sc), an abnormal conformational isoform of the prion protein, which, like its normal cellular precursor (PrP(C)), has a GPI (glycosylphosphatidylinositol) anchor at the C-terminus. To date, elucidating the role of this anchor on the infectivity of prion preparations has not been possible because of the resistance of PrP(Sc) to the activity of PI-PLC (phosphoinositide-specific phospholipase C), an enzyme which removes the GPI moiety from PrP(C). Removal of the GPI anchor from PrP(Sc) requires denaturation before treatment with PI-PLC, a process that also abolishes infectivity. To circumvent this problem, we have removed the GPI anchor from PrP(Sc) in RML (Rocky Mountain Laboratory)-prion-infected murine brain homogenate using the aspartic endoprotease cathepsin D. This enzyme eliminates a short sequence at the C-terminal end of PrP to which the GPI anchor is attached. We found that this modification has no effect (i) on an in vitro amplification model of PrP(Sc), (ii) on the prion titre as determined by a highly sensitive N2a-cell based bioassay, or (iii) in a mouse bioassay. These results show that the GPI anchor has little or no role in either the propagation of PrP(Sc) or on prion infectivity. PMID:16441239

  3. Removal of the glycosylphosphatidylinositol anchor from PrPSc by cathepsin D does not reduce prion infectivity

    Lewis, Patrick A.; Properzi, Francesca; Prodromidou, Kanella; Clarke, Anthony R.; Collinge, John; Jackson, Graham S.

    2006-01-01

    According to the protein-only hypothesis of prion propagation, prions are composed principally of PrPSc, an abnormal conformational isoform of the prion protein, which, like its normal cellular precursor (PrPC), has a GPI (glycosylphosphatidylinositol) anchor at the C-terminus. To date, elucidating the role of this anchor on the infectivity of prion preparations has not been possible because of the resistance of PrPSc to the activity of PI-PLC (phosphoinositide-specific phospholipase C), an enzyme which removes the GPI moiety from PrPC. Removal of the GPI anchor from PrPSc requires denaturation before treatment with PI-PLC, a process that also abolishes infectivity. To circumvent this problem, we have removed the GPI anchor from PrPSc in RML (Rocky Mountain Laboratory)-prion-infected murine brain homogenate using the aspartic endoprotease cathepsin D. This enzyme eliminates a short sequence at the C-terminal end of PrP to which the GPI anchor is attached. We found that this modification has no effect (i) on an in vitro amplification model of PrPSc, (ii) on the prion titre as determined by a highly sensitive N2a-cell based bioassay, or (iii) in a mouse bioassay. These results show that the GPI anchor has little or no role in either the propagation of PrPSc or on prion infectivity. PMID:16441239

  4. Geodesic defect anchoring on nematic shells.

    Mirantsev, Leonid V; Sonnet, André M; Virga, Epifanio G

    2012-08-01

    Nematic shells are colloidal particles coated with nematic liquid crystal molecules, which may freely glide and rotate on the colloid's surface while keeping their long axis on the local tangent plane. Molecular dynamics simulations on a nanoscopic spherical shell indicate that under appropriate adhesion conditions for the molecules on the equator, the equilibrium nematic texture exhibits at each pole a pair of +1/2 defects so close to one another to be treated as one +1 defect. Spirals connect the polar defects, though the continuum limit of the interaction potential would not feature any elastic anisotropy. A molecular averaging justifies an anchoring defect energy that feels the geodesics emanating from the defect. All our observations are explained by such a geodesic anchoring, which vanishes on flat manifolds. PMID:23005713

  5. Recent Study of Drag Embedment Plate Anchors in China

    Haixiao Liu

    2012-01-01

    Experimental and theoretical studies of drag embedment plate anchors recently carried out in Tianjin University are summarized in this research paper,which involve a series of important topics relevant to the study of drag anchors.The techniques for measuring the trajectory and movement direction of drag anchors in soils,the techniques for measuring the moving embedment point and reverse catenary shape of the embedded drag line,the penetration mechanism and kinematic behavior of drag anchors,the ultimate embedment depth of drag anchors,the movement direction of the anchor with an arbitrary fluke section,the reverse catenary properties of the embedded drag line,the interactional properties between drag anchor and installation line,the kinematic model of drag anchors in seabed soils,and the analytical method for predicting the anchor trajectory in soils will all be examined.The present work remarkably reduces the uncertainties in design and analysis of drag embedment plate anchors,and is beneficial to improving the application of this new type of drag anchor in offshore engineering.

  6. Anchor Toolkit - a secure mobile agent system

    Mudumbai, Srilekha S.; Johnston, William; Essiari, Abdelilah

    2008-01-01

    Mobile agent technology facilitates intelligent operation in software systems with less human interaction. Major challenge to deployment of mobile agents include secure transmission of agents and preventing unauthorized access to resources between interacting systems, as either hosts, or agents, or both can act maliciously. The Anchor toolkit, designed by LBNL, handles the transmission and secure management of mobile agents in a heterogeneous distributed computing environment. It provide...

  7. Anchoring in a novel bimanual coordination pattern.

    Maslovat, Dana; Lam, Melanie Y; Brunke, Kirstin M; Chua, Romeo; Franks, Ian M

    2009-02-01

    Anchoring in cyclical movements has been defined as regions of reduced spatial or temporal variability [Beek, P. J. (1989). Juggling dynamics. PhD thesis. Amsterdam: Free University Press] that are typically found at movement reversal points. For in-phase and anti-phase movements, synchronizing reversal points with a metronome pulse has resulted in decreased anchor point variability and increased pattern stability [Byblow, W. D., Carson, R. G., & Goodman, D. (1994). Expressions of asymmetries and anchoring in bimanual coordination. Human Movement Science, 13, 3-28; Fink, P. W., Foo, P., Jirsa, V. K., & Kelso, J. A. S. (2000). Local and global stabilization of coordination by sensory information. Experimental Brain Research, 134, 9-20]. The present experiment examined anchoring during acquisition, retention, and transfer of a 90 degrees phase-offset continuous bimanual coordination pattern (whereby the right limb lags the left limb by one quarter cycle), involving horizontal flexion about the elbow. Three metronome synchronization strategies were imposed: participants either synchronized maximal flexion of the right arm (i.e., single metronome), both flexion and extension of the right arm (i.e., double metronome within-limb), or flexion of each arm (i.e., double metronome between-limb) to an auditory metronome. In contrast to simpler in-phase and anti-phase movements, synchronization of additional reversal points to the metronome did not reduce reversal point variability or increase pattern stability. Furthermore, practicing under different metronome synchronization strategies did not appear to have a significant effect on the rate of acquisition of the pattern. PMID:18842313

  8. Molecular Conductance: Chemical Trends of Anchoring Groups

    Ke, San-Huang; Baranger, Harold U.; Yang, Weitao

    2004-01-01

    Combining density functional theory calculations for molecular electronic structure with a Green function method for electron transport, we calculate from first principles the molecular conductance of benzene connected to two Au leads through different anchoring atoms -- S, Se, and Te. The relaxed atomic structure of the contact, different lead orientations, and different adsorption sites are fully considered. We find that the molecule-lead coupling, electron transfer, and conductance all dep...

  9. Anchoring Heuristic and the Equity Premium Puzzle

    Siddiqi, Hammad

    2015-01-01

    What happens when the anchoring and adjustment heuristic of Tversky and Kahneman (1974) is incorporated in the standard consumption-based capital asset pricing model (CCAPM)? The surprising finding is that it not only resolves the high equity-premium and low risk-free rate puzzles with a low risk-aversion coefficient, but also provides a unified framework for understanding countercyclical equity-premium, excess volatility, size, value, and momentum effects, and abnormal returns and volatiliti...

  10. Crystal structure of GAP50, the anchor of the invasion machinery in the inner membrane complex of Plasmodium falciparum

    van der Bosch, Jürgen; Paige, Matthew H.; Vaidya, Akhil; Bergman, Lawrence; Hol, Wim G. J.

    2012-01-01

    The glideosome associated protein GAP50 is an essential protein in apicomplexan parasites such as Plasmodium, Toxoplasma and Cryptosporidium, several species of which are important human pathogens. The 44.6 kDa protein is part of a multi-protein complex known as the invasion machinery or glideosome, which is required for cell invasion and substrate gliding motility empowered by an actin-myosin motor. GAP50 is anchored through its C-terminal transmembrane helix into the inner membrane complex ...

  11. The Analysis Stability of Anchor Retaining Wall

    The construction of anchored retaining walls reach every day in the field of Civil Engineering especially in public works. Their dimensioning and stability are the axes of research for geotechnical. The rule is to reduce the active forces of the slide and increase the effective normal stress on the rupture surface. So that, we anchored tied-back (constituted by steel cables) in the stable ground located under the failure surface and we apply at the top a traction force. This effort can be distributed over the ground surface by means of small plates or massive reinforced concrete. The study of the stability of anchored retaining wall was also performed by using software GEO4. Many cases can be solved using analytical solutions available in the group GEO4 program, but for our standard model solution studied analytically proved unsatisfactory so we used a numerical analysis based on the method of finite element in this program. The results obtained by numerical study were interpreted to identify the precision numerical predictions. Moreover these methods were useful and economics in the realization of reinforced slopes by tied-buck. (author)

  12. Structural basis for membrane anchoring of HIV-1 envelope spike.

    Dev, Jyoti; Park, Donghyun; Fu, Qingshan; Chen, Jia; Ha, Heather Jiwon; Ghantous, Fadi; Herrmann, Tobias; Chang, Weiting; Liu, Zhijun; Frey, Gary; Seaman, Michael S; Chen, Bing; Chou, James J

    2016-07-01

    HIV-1 envelope spike (Env) is a type I membrane protein that mediates viral entry. We used nuclear magnetic resonance to determine an atomic structure of the transmembrane (TM) domain of HIV-1 Env reconstituted in bicelles that mimic a lipid bilayer. The TM forms a well-ordered trimer that protects a conserved membrane-embedded arginine. An amino-terminal coiled-coil and a carboxyl-terminal hydrophilic core stabilize the trimer. Individual mutations of conserved residues did not disrupt the TM trimer and minimally affected membrane fusion and infectivity. Major changes in the hydrophilic core, however, altered the antibody sensitivity of Env. These results show how a TM domain anchors, stabilizes, and modulates a viral envelope spike and suggest that its influence on Env conformation is an important consideration for HIV-1 immunogen design. PMID:27338706

  13. Robotic Ankle for Omnidirectional Rock Anchors

    Parness, Aaron; Frost, Matthew; Thatte, Nitish

    2013-01-01

    Future robotic exploration of near-Earth asteroids and the vertical and inverted rock walls of lava caves and cliff faces on Mars and other planetary bodies would require a method of gripping their rocky surfaces to allow mobility without gravitational assistance. In order to successfully navigate this terrain and drill for samples, the grippers must be able to produce anchoring forces in excess of 100 N. Additionally, the grippers must be able to support the inertial forces of a moving robot, as well gravitational forces for demonstrations on Earth. One possible solution would be to use microspine arrays to anchor to rock surfaces and provide the necessary load-bearing abilities for robotic exploration of asteroids. Microspine arrays comprise dozens of small steel hooks supported on individual suspensions. When these arrays are dragged along a rock surface, the steel hooks engage with asperities and holes on the surface. The suspensions allow for individual hooks to engage with asperities while the remaining hooks continue to drag along the surface. This ensures that the maximum possible number of hooks engage with the surface, thereby increasing the load-bearing abilities of the gripper. Using the microspine array grippers described above as the end-effectors of a robot would allow it to traverse terrain previously unreachable by traditional wheeled robots. Furthermore, microspine-gripping robots that can perch on cliffs or rocky walls could enable a new class of persistent surveillance devices for military applications. In order to interface these microspine grippers with a legged robot, an ankle is needed that can robotically actuate the gripper, as well as allow it to conform to the large-scale irregularities in the rock. The anchor serves three main purposes: deploy and release the anchor, conform to roughness or misalignment with the surface, and cancel out any moments about the anchor that could cause unintentional detachment. The ankle design contains a

  14. Removal of the glycosylphosphatidylinositol anchor from PrPSc by cathepsin D does not reduce prion infectivity

    Lewis, P. A.; Properzi, F.; Prodromidou, K.; Clarke, A R; Collinge, J.; Jackson, G S

    2006-01-01

    According to the protein-only hypothesis of prion propagation, prions are composed principally of PrPSc, an abnormal conformational isoform of the prion protein, which, like its normal cellular precursor (PrPC), has a GPI (glycosylphosphatidylinositol) anchor at the C-terminus. To date, elucidating the role of this anchor on the infectivity of prion preparations has not been possible because of the resistance of PrPSc to the activity of PI-PLC (phosphoinositide specific phospholipase C), an e...

  15. Protein (Viridiplantae): 308812183 [

    Full Text Available XP_003083399.1 33090:12970 3041:5897 1035538:4613 13792:4613 70447:1628 70448:5196 Glycosylphosp ... hatidylinositol anchor synthesis ... protein (ISS) Ostreococcus tauri MSARRASFQSRFNDSSQ ...

  16. Allosteric inhibition of Aurora-A kinase by a synthetic vNAR domain.

    Burgess, Selena G; Oleksy, Arkadiusz; Cavazza, Tommaso; Richards, Mark W; Vernos, Isabelle; Matthews, David; Bayliss, Richard

    2016-07-01

    The vast majority of clinically approved protein kinase inhibitors target the ATP-binding pocket directly. Consequently, many inhibitors have broad selectivity profiles and most have significant off-target effects. Allosteric inhibitors are generally more selective, but are difficult to identify because allosteric binding sites are often unknown or poorly characterized. Aurora-A is activated through binding of TPX2 to an allosteric site on the kinase catalytic domain, and this knowledge could be exploited to generate an inhibitor. Here, we generated an allosteric inhibitor of Aurora-A kinase based on a synthetic, vNAR single domain scaffold, vNAR-D01. Biochemical studies and a crystal structure of the Aurora-A/vNAR-D01 complex show that the vNAR domain overlaps with the TPX2 binding site. In contrast with the binding of TPX2, which stabilizes an active conformation of the kinase, binding of the vNAR domain stabilizes an inactive conformation, in which the αC-helix is distorted, the canonical Lys-Glu salt bridge is broken and the regulatory (R-) spine is disrupted by an additional hydrophobic side chain from the activation loop. These studies illustrate how single domain antibodies can be used to characterize the regulatory mechanisms of kinases and provide a rational basis for structure-guided design of allosteric Aurora-A kinase inhibitors. PMID:27411893

  17. Dynamic performance of concrete undercut anchors for Nuclear Power Plants

    Graphical abstract: - Highlights: • Behavior of undercut anchors under dynamic actions simulating earthquakes. • First high frequency load and crack cycling tests on installed concrete anchors ever. • Comprehensive review of anchor qualification for Nuclear Power Plants. - Abstract: Post-installed anchors are widely used for structural and nonstructural connections to concrete. In many countries, concrete anchors used for Nuclear Power Plants have to be qualified to ensure reliable behavior even under extreme conditions. The tests required for qualification of concrete anchors are carried out at quasi-static loading rates well below the rates to be expected for dynamic actions deriving from earthquakes, airplane impacts or explosions. To investigate potentially beneficial effects of high loading rates and cycling frequencies, performance tests on installed undercut anchors were conducted. After introductory notes on anchor technology and a comprehensive literature review, this paper discusses the qualification of anchors for Nuclear Power Plants and the testing carried out to quantify experimentally the effects of dynamic actions on the load–displacement behavior of undercut anchors

  18. Analysis of Glenoid Inter-anchor Distance with an All-Suture Anchor System

    Kramer, Jonathan; Robinson, Sean; Dutton, Pascual; Dickinson, Ephraim; Rodriguez, John Paul; Camisa, William; Leasure, Jeremi M.; Montgomery, William H.

    2016-01-01

    Objectives: Arthroscopic shoulder stabilization using suture anchors are commonly used techniques. More recently developed all-suture systems employ smaller diameter anchors, which increase repair contact area and allow greater placement density on narrow surfaces such as the glenoid. Our goal is investigate the strength characteristics of various inter-anchor distances in a human glenoid model. Methods: Twelve fresh-frozen human cadaveric glenoids were potted after the labrum was excised. The glenoids were then implanted with 1.4 mm all-suture anchors (Juggerknot, Biomet, Warsaw, IN) at varying inter-anchor distances. Anchors were implanted adjacent to one another or at 2 mm, 3 mm, or 5 mm distances using a template with pre-drilled holes. The glenoids were then underwent single cycle pullout testing using a test frame (Instron 8521, Instron Inc., Norwood, MA). A 5 N preload was applied to the construct and the actuator was driven away from the shoulder at a rate of 12.5mm/s as seen in Figure 1. Force and displacement were collected from the test frame actuator at a rate of 500 Hz. The primary outcomes were failure strength and stiffness. Stiffness was calculated from the initial linear region of the force displacement curve. Failure strength was defined as the first local maximum inflection point in the force displacement curve. Results: During load to fail testing, all but three of the specimens had both anchors pull out of the glenoid. The other mode of failure included one or both of the sutures failing. Stiffness was 13.52 ± 3.8, 17.97 ± 5.02, 17.59 ± 4.65 and 18.95 ± 4.67 N/mm for the adjacent, 2 mm, 3 mm and 5 mm treatment groups as shown Table 1. The adjacent group had a significantly lower stiffness compared to the other treatment groups. Failure strength was 48.68 ± 20.64, 76.16 ± 23.78, 73.19 ± 35.83 and 87.04 ± 34.67 N for the adjacent, 2 mm, 3 mm and 5 mm treatment groups as shown in Table 1. The adjacent group had a significantly lower

  19. Adsorption phenomena and anchoring energy in nematic liquid crystals

    Barbero, Giovanni

    2005-01-01

    Despite the large quantity of phenomenological information concerning the bulk properties of nematic phase liquid crystals, little is understood about the origin of the surface energy, particularly the surface, interfacial, and anchoring properties of liquid crystals that affect the performance of liquid crystal devices. Self-contained and unique, Adsorption Phenomena and Anchoring Energy in Nematic Liquid Crystals provides an account of new and established results spanning three decades of research into the problems of anchoring energy and adsorption phenomena in liquid crystals.The book contains a detailed discussion of the origin and possible sources of anchoring energy in nematic liquid crystals, emphasizing the dielectric contribution to the anchoring energy in particular. Beginning with fundamental surface and anchoring properties of liquid crystals and the definition of the nematic phase, the authors explain how selective ion adsorption, dielectric energy density, thickness dependence, and bias voltage...

  20. The bone-anchored hearing aid

    Foghsgaard, Søren

    2014-01-01

    The bone-anchored hearing aid (Baha) was introduced in 1977 by Tjellström and colleagues and has now been used clinically for over 30 years. Generally, the outcomes are good, and several studies have shown improved audiological- and quality of life outcomes. The principle of the Baha is, that sound...... vibrations are led directly to the inner ear via the mastoid bone, bypassing the middle ear. This is achieved via an osseointegrated implant and a skin-penetrating abutment. Studies report high success rates and a majority of complications as typically minor in nature....

  1. Anchor Toolkit - a secure mobile agent system

    Mudumbai, Srilekha S.; Johnston, William; Essiari, Abdelilah

    1999-05-19

    Mobile agent technology facilitates intelligent operation insoftware systems with less human interaction. Major challenge todeployment of mobile agents include secure transmission of agents andpreventing unauthorized access to resources between interacting systems,as either hosts, or agents, or both can act maliciously. The Anchortoolkit, designed by LBNL, handles the transmission and secure managementof mobile agents in a heterogeneous distributed computing environment. Itprovides users with the option of incorporating their security managers.This paper concentrates on the architecture, features, access control anddeployment of Anchor toolkit. Application of this toolkit in a securedistributed CVS environment is discussed as a case study.

  2. Starting point anchoring effects in choice experiments

    Ladenburg, Jacob; Olsen, Søren Bøye

    Anchoring is acknowledged as a potential source of considerable bias in Dichotomous Choice Contingent Valuation studies. Recently, another stated preference method known as Choice Experiments has gained in popularity as well as the number of applied studies. However, as the elicitation of...... subjectivity in the present study is gender dependent, pointing towards, that female respondents are prone to be affected by the price levels employed. Male respondents, on the other hand, are not sensitive towards these prices levels. Overall, this implicates that female respondents, when employing a low...

  3. The impact of anchoring bias in the UK equity market

    Koskinen, Matti

    2013-01-01

    PURPOSE OF THE STUDY The purpose of this thesis is to study how the anchoring bias, a cognitive bias, affects equity analysts' forecasts and what kind of implications this bias has for investors and managers of listed companies in the UK stock market. I test whether industry median forecast earnings per share can work as an anchor when analysts are estimating companies' future earnings and how this potential anchoring affects forecast errors, future stock returns, earnings surprises and s...

  4. Nominal Anchor Exchange Rate Policies as a Domestic Distortion

    Anne O. Krueger

    1997-01-01

    This paper analyzes a nominal anchor exchange rate policy as a domestic distortion, in the tradition of international trade theory. It is shown that, in addition to the problems of sustainability and exit pinpointed in the exchange rate literature, a nominal anchor exchange rate policy, while in force, drives a wedge between the domestic and the international intertemporal marginal rates of substitution. The welfare cost of the Mexican use of the nominal anchor exchange rate policy prior to D...

  5. Decoding Cytoskeleton-Anchored and Non-Anchored Receptors from Single-Cell Adhesion Force Data.

    Sariisik, Ediz; Popov, Cvetan; Müller, Jochen P; Docheva, Denitsa; Clausen-Schaumann, Hauke; Benoit, Martin

    2015-10-01

    Complementary to parameters established for cell-adhesion force curve analysis, we evaluated the slope before a force step together with the distance from the surface at which the step occurs and visualized the result in a two-dimensional density plot. This new tool allows detachment steps of long membrane tethers to be distinguished from shorter jumplike force steps, which are typical for cytoskeleton-anchored bonds. A prostate cancer cell line (PC3) immobilized on an atomic-force-microscopy sensor interacted with three different substrates: collagen-I (Col-I), bovine serum albumin, and a monolayer of bone marrow-derived stem cells (SCP1). To address PC3 cells' predominant Col-I binding molecules, an antibody-blocking β1-integrin was used. Untreated PC3 cells on Col-I or SCP1 cells, which express Col-I, predominantly showed jumps in their force curves, while PC3 cells on bovine-serum-albumin- and antibody-treated PC3 cells showed long membrane tethers. The probability density plots thus revealed that β1-integrin-specific interactions are predominately anchored to the cytoskeleton, while the nonspecific interactions are mainly membrane-anchored. Experiments with latrunculin-A-treated PC3 cells corroborated these observations. The plots thus reveal details of the anchoring of bonds to the cell and provide a better understanding of receptor-ligand interactions. PMID:26445433

  6. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either 3H-fatty acids or [3H]ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the 3H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of [3H]ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from 3H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the 3H-fatty acid and the [3H]ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the [3H]ethanolamine label from the purified alkaline phosphatase. The 3H radioactivity in alkaline phosphatase purified from [3H]ethanolamine-labeled cells comigrated with authentic [3H]ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the 3H-fatty acid and [3H]ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase

  7. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Jemmerson, R.; Low, M.G.

    1987-09-08

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either /sup 3/H-fatty acids or (/sup 3/H)ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the /sup 3/H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of (/sup 3/H)ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from /sup 3/H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the /sup 3/H-fatty acid and the (/sup 3/H)ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the (/sup 3/H)ethanolamine label from the purified alkaline phosphatase. The /sup 3/H radioactivity in alkaline phosphatase purified from (/sup 3/H)ethanolamine-labeled cells comigrated with authentic (/sup 3/H)ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the /sup 3/H-fatty acid and (/sup 3/H)ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase.

  8. Illusion of Relevance: Anchoring in Economic and Financial Knowledge

    Andrey Kudryavtse

    2010-12-01

    Full Text Available Contemporary research documents various psychological aspects of economic thought and decision-making. The main goal of our study is to analyze the role of the anchoring (bias(Tversky and Kahneman, 1974 in perceiving economic and financial information, and, in particular, the effect of perceived relevance of the anchors on the degree of the bias. Anchoring bias refers to people's tendency to form their estimates for different categories, starting from a particular available, and often irrelevant, value and insufficiently adjusting their final judgments from this starting value. We carry out an experiment involving a group of MBA students, asking them to recall a number of recent economic and financial indicators (stock and bond market index returns, rates of inflation, currency exchange rates, etc., with half of the participants receiving actual information about some unrelated indicators (anchors, before answering the questions. We document that significant anchoring bias is exhibited, on average, for each of our experimental questions and by vast majority of the participants, the degree of the bias being higher for women and older participants. Furthermore, in the context of the effect of anchors' perceived relevance, we find that people exhibit significantly stronger bias in cases when the anchor is presented as a similar category. Our findings indicate that if, at least at the first glance, an anchor bears some similarity to the target question, then the resulting answer may be more assimilated towards the anchor

  9. Secreted form of the human interleukin 2 receptor encoded by an anchor minus cDNA

    The DNA sequence encoding all of the putative intracytoplasmic domain and most of the trans-membrane domain of the huma IL 2 receptor was detected from a full length receptor cDNA. After expression in mouse L cells, the resultant anchor minus cDNA was found to direct the synthesis of a secreted rather than membrane-associated form of the IL 2 receptor. The secreted receptor protein (44,000 to 46,000 M/sub r/) retained the capacity to bind both IL 2 and the monoclonal anti-Tac antibody, as evidenced by (1) retention on IL 2 and anti-Tac affinity columns, (2) inhibition of [3H]-antiTac binding to HUT 102B2 cells, and (3) partial inhibition of IL 2-induced CTLL proliferation. Removal of these domains from the IL 2 receptor did not alter the post-translational processing or rate of export of the truncated receptor protein. These data confirm the proposed membrane orientation of the IL 2 receptor (NH2 terminus out, COOH terminus in) and underscore the anchoring function of this carboxy terminal receptor segment. The availability of such anchor minus receptor cDNA constructs may facilitate purification of large quantities of receptor protein for further analysis of receptor structure, valency, and localization of the IL 2 binding site(s)

  10. Decay accelerating factor (DAF) is anchored to membranes by a C-terminal glycolipid

    Purified 70 kDa membrane (m) DAF incorporates into cells when added in vitro. A 2 kDa smaller DAF form which functions extrinsically like C4bp but is unable to incorporate can be isolated from urine (u). Because of common deficits of mDAF and acetylcholinesterase (AChE) in erythrocytes (E) of patients with paroxysmal nocturnal hemoglobinuria (PNH), mDAF was analyzed for a O-terminal glycolipid membrane anchor similar to that in E AChE. Incubation of E with phosphatidylinositol-specific phospholipase C, an enzyme which cleaves a similar glycolipid anchor in trypanosome variant surface glycoproteins (mfVSGs), released 20% of the DAF antigen. The released DAF species resembled uDAF in size, extrinsic model of C4b2a decay, and lack of hydrophobicity. Reductive radiomethylation of mDAF with [14C]HCHO and NaCNBH3 revealed ethanolamine and glucosamine in proportions similar to those in the E AChE glycolipid anchor. Papain cleavage of radiomethylated mDAF released the labeled ethanolamine and glucosamine in small O-terminal fragments from the residual DAF that retained N-terminal Asp. Following labeling of the anchors of mDAF and E AChE with the lipophilic photoreagent 3-trifluoromethyl-3-(m-[125I]iodophenyl)diazirine, cleavage at the glucosamine residue by deamination quantitatively released the label from both proteins. Biosynthetic labeling of Hela cells with [3H]ethanolamine resulted in rapid 3H incorporation into both 48 kDa proDAF and 70 kDa mDAF. These data indicate that mDAF is anchored by a glycolipid similar to that in E AChE, mfVSGs and Thy-1 antigen and raise the possibility that a defect in the assembly or attachment of this structure could account for the deficits of mDAF and E AChE in PNH

  11. Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells

    von Nickisch-Rosenegk Markus

    2012-01-01

    Full Text Available Abstract The need to functionalize cell membranes in a directed way for specific applications as single cell arrays or to force close cell-to-cell contact for artificial intercellular interaction and/or induction concerning stem cell manipulation or in general to have a tool for membrane and cell surface-associated processes, we envisaged a neutral inactive membrane anchor for extracellular entities to facillitate the above mentioned functionalities. The silent Duffy antigen/receptor for chemokines (DARC is a receptor-like membrane protein of erythrocytes and mediates no cell transduction not at least regarding a missing or truncated G-loop and therefore it seemed to be the candidate for our cell membrane anchor. We isolated the genetic information of DARC from human genomic DNA and cloned it in a mammalian cell line as a fusion protein via a suitable plasmid vector. In this report we demonstrate that the human plasma membrane protein DARC can be used as an artificial anchor molecule in cell surface engineering applications. We constructed the fusion protein SNAP-tag-DARC, consisting of DARC and the self-labeling protein tag SNAP-tag® (Covalys. The SNAP-tag® served as an example for a molecular-technological developed protein that is artificially attached to the extracellular side of the plasma membrane through our DARC-anchor. SnapTag should serve as an example for any extracellular entity and was easy to detect by a commercial detection system. The synthesis of SNAP-tag-DARC, its correct incorporation into the cell membrane and the functionality of the SNAP-tag® were verified by RT-PCR, Western blotting and confocal fluorescence microscopy and showed the desired functionality as an membrane anchor for an extracellular application entity.

  12. An androgenic gland membrane-anchored gene associated with the crustacean insulin-like androgenic gland hormone.

    Rosen, Ohad; Manor, Rivka; Weil, Simy; Aflalo, Eliahu D; Bakhrat, Anna; Abdu, Uri; Sagi, Amir

    2013-06-01

    Crustacean male sexual differentiation is governed by the androgenic gland (AG) and specifically by the secreted insulin-like AG hormone (IAG), thus far identified in several decapod species including the Australian red claw crayfish Cherax quadricarinatus (termed Cq-IAG). While a few insulin-like AG genes have been identified in crustaceans, other AG-specific genes have not been documented until now. In the present study, we describe the recent identification of a non-IAG AG-specific transcript obtained from the C. quadricarinatus AG cDNA library. This transcript, termed C. quadricarinatus membrane-anchored AG-specific factor (Cq-MAG), was fully sequenced and found to encode a putative product of 189 amino acids including a signal anchoring peptide. Expression of a recombinant GFP fusion protein lacking the signal anchor encoding sequence dramatically affected recombinant protein localization pattern. While the expression of the deleterious fusion protein was observed throughout most of the cell, the native GFP::Cq-MAG fusion protein was observed mainly surrounding the periphery of the nucleus, demonstrating an endoplasmic reticulum (ER)-like localization pattern. Moreover, co-expression of the wild-type Cq-MAG (fused to GFP) and the Cq-IAG hormone revealed that these peptides indeed co-localize. This study is the first to report a protein specifically associated with the insulin-like AG hormone in addition to the finding of another AG-specific transcript in crustaceans. Previous knowledge suggests that insulin/insulin-like factor secretion involves tissue-specific transcripts and membrane-anchored proteins. In this regard, Cq-MAG's tissue specificity, anchoring properties and intracellular co-localization with Cq-IAG suggest that it may play a role in the processing and secretion of this insulin-like AG hormone. PMID:23470660

  13. Two novel, putatively cell wall-associated and glycosylphosphatidylinositol-anchored α-glucanotransferase enzymes of Aspergillus niger

    Kaaij, R.M. van der; Yuan, X.L.; Franken, A.; Ram, A.F.J.; Punt, P.J.; Maarel, M.J.E.C. van der; Dijkhuizen, L.

    2007-01-01

    In the genome sequence of Aspergillus niger CBS 513.88, three genes were identified with high similarity to fungal α-amylases. The protein sequences derived from these genes were different in two ways from all described fungal α-amylases: they were predicted to be glycosylphosphatidylinositol anchor

  14. Cellular receptor for urokinase plasminogen activator. Carboxyl-terminal processing and membrane anchoring by glycosyl-phosphatidylinositol

    Ploug, M; Rønne, E; Behrendt, N;

    1991-01-01

    The cellular receptor for human urokinase-type plasminogen activator (u-PAR) is shown by several independent criteria to be a true member of a family of integral membrane proteins, anchored to the plasma membrane exclusively by a COOH-terminal glycosyl-phosphatidylinositol moiety. 1) Amino acid a...

  15. 30 CFR 57.9311 - Anchoring stationary sizing devices.

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Anchoring stationary sizing devices. 57.9311 Section 57.9311 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR METAL AND... Dumping Sites § 57.9311 Anchoring stationary sizing devices. Grizzlies and other stationary sizing...

  16. 30 CFR 56.9311 - Anchoring stationary sizing devices.

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Anchoring stationary sizing devices. 56.9311 Section 56.9311 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR METAL AND... Dumping Sites § 56.9311 Anchoring stationary sizing devices. Grizzlies and other stationary sizing...

  17. Understanding Rasch Measurement: Partial Credit Model and Pivot Anchoring.

    Bode, Rita K.

    2001-01-01

    Describes the Rasch measurement partial credit model, what it is, how it differs from other Rasch models, and when and how to use it. Also describes the calibration of instruments with increasingly complex items. Explains pivot anchoring and illustrates its use and describes the effect of pivot anchoring on step calibrations, item hierarchy, and…

  18. Sperm protein 17 is expressed in the sperm fibrous sheath

    Albani Elena

    2009-07-01

    Full Text Available Abstract Background Sperm protein 17 (Sp17 is a highly conserved mammalian protein characterized in rabbit, mouse, monkey, baboon, macaque, human testis and spermatozoa. mRNA encoding Sp17 has been detected in a range of murine and human somatic tissues. It was also recognized in two myeloma cell lines and in neoplastic cells from patients with multiple myeloma and ovarian carcinoma. These data all indicate that Sp17 is widely distributed in humans, expressed not only in germinal cells and in a variety of somatic tissues, but also in neoplastic cells of unrelated origin. Methods Sp17 expression was analyzed by immunocytochemistry and transmission electron microscopy on spermatozoa. Results Here, we demonstrate the ultrastructural localization of human Sp17 throughout the spermatozoa flagellar fibrous sheath, and its presence in spermatozoa during in vitro states from their ejaculation to the oocyte fertilization. Conclusion These findings suggest a possible role of Sp17 in regulating sperm maturation, capacitation, acrosomal reaction and interactions with the oocyte zona pellucida during the fertilization process. Further, the high degree of sequence conservation throughout its N-terminal half, and the presence of an A-kinase anchoring protein (AKAP-binding motif within this region, suggest that Sp17 might play a regulatory role in a protein kinase A-independent AKAP complex in both germinal and somatic cells.

  19. Breaking anchored droplets in a microfluidic Hele-Shaw cell

    Amselem, Gabriel; Gallaire, François; Baroud, Charles N

    2015-01-01

    We study microfluidic self digitization in Hele-Shaw cells using pancake droplets anchored to surface tension traps. We show that above a critical flow rate, large anchored droplets break up to form two daughter droplets, one of which remains in the anchor. Below the critical flow velocity for breakup the shape of the anchored drop is given by an elastica equation that depends on the capillary number of the outer fluid. As the velocity crosses the critical value, the equation stops admitting a solution that satisfies the boundary conditions; the drop breaks up in spite of the neck still having finite width. A similar breaking event also takes place between the holes of an array of anchors, which we use to produce a 2D array of stationary drops in situ.

  20. Astral microtubule pivoting promotes their search for cortical anchor sites during mitosis in budding yeast.

    Stephan Baumgärtner

    Full Text Available Positioning of the mitotic spindle is crucial for proper cell division. In the budding yeast Saccharomyces cerevisiae, two mechanisms contribute to spindle positioning. In the Kar9 pathway, astral microtubules emanating from the daughter-bound spindle pole body interact via the linker protein Kar9 with the myosin Myo2, which moves the microtubule along the actin cables towards the neck. In the dynein pathway, astral microtubules off-load dynein onto the cortical anchor protein Num1, which is followed by dynein pulling on the spindle. Yet, the mechanism by which microtubules target cortical anchor sites is unknown. Here we quantify the pivoting motion of astral microtubules around the spindle pole bodies, which occurs during spindle translocation towards the neck and through the neck. We show that this pivoting is largely driven by the Kar9 pathway. The microtubules emanating from the daughter-bound spindle pole body pivot faster than those at the mother-bound spindle pole body. The Kar9 pathway reduces the time needed for an astral microtubule inside the daughter cell to start pulling on the spindle. Thus, we propose a new role for microtubule pivoting: By pivoting around the spindle pole body, microtubules explore the space laterally, which helps them search for cortical anchor sites in the context of spindle positioning in budding yeast.

  1. 利用枯草芽孢衣壳蛋白表面展示β-半乳糖苷酶%Functional Display of β-galactosidase on the Spore Surface of Bacillus subtilis Using Spore Coat Protein as Anchor Motif

    王贺; 杨瑞金; 华霄; 赵伟; 张文斌

    2012-01-01

    分别将枯草芽孢杆菌(Bacillussubtilis 168)芽孢衣壳蛋白CotB、CotC、CotG和CotX的启动子和编码序列与来自嗜热脂肪芽孢杆菌(BacillusstearothermophilusIAMll001)的β-半乳糖苷酶基因bgaB进行重组,构建融合表达cotB—bgaB、eotC—bgaB、eotG—bgaB和eotX—bgaB的整合型重组质粒。将4种重组质粒分别转入枯草芽孢杆菌Bacillussubtilis168(trp。),获得了能在芽孢表面展示的重组菌株PB701、PB702、PB703和PB704。经Westernblot检测,4种重组菌株均表达了预期分子量的融合蛋白,初步表明β-半乳糖苷酶被锚定在重组菌株的芽孢表面。以oNPG为底物测定4种重组菌株芽孢表面展示β-半乳糖苷酶的水解能力,得到的酶活分别为0.14、0.06、0.22和0.20U/mL。%In this work, we developed an efficient spore display system that a model protein β-galactosidase was anchored on the spore surface of Bacillus subtilis 168 based on the use of spore coat proteins. The PCR-amplifying cotB, cotC, cotG and cotX were ligated with pMD-19T and digested with XbaI and KpnI, and then subcloned into vector pJS700a previously digested with the same two restriction enzymes, finally resulted in the plasmids pJSB, pJSC, pJSG and pJSX. To construct the gene fusions, the bgaB from Bacillus stearothermophilus IAMll001 was cloned into the KpnI and EcoRI sites of plasmid pJSB, pJSC, pJS G and pJSX to generate generating the plasmids pJSBB, pJSCB, pJSGB and pJSXB,respectively After linearization with BgllI restriction endonuclease, the four re- combinant integrative plasmids were transformed into B. subtilis 168 to yield the recombinant strain PB701, PB702, PB703 and PB704,respectively. Results from Western blot analysis showed that the fusion protein was immobilized on the spore surface. Using oNPG as substrate, the enzyme activity of spore-displaying β-galactosidase was assayed and they were 0.14, 0.06, 0.22 and 0.20 U/mL for PB701, PB702, PB

  2. Peptide Anchor for Folate-Targeted Liposomal Delivery.

    Nogueira, Eugénia; Mangialavori, Irene C; Loureiro, Ana; Azoia, Nuno G; Sárria, Marisa P; Nogueira, Patrícia; Freitas, Jaime; Härmark, Johan; Shimanovich, Ulyana; Rollett, Alexandra; Lacroix, Ghislaine; Bernardes, Gonçalo J L; Guebitz, Georg; Hebert, Hans; Moreira, Alexandra; Carmo, Alexandre M; Rossi, Juan Pablo F C; Gomes, Andreia C; Preto, Ana; Cavaco-Paulo, Artur

    2015-09-14

    Specific folate receptors are abundantly overexpressed in chronically activated macrophages and in most cancer cells. Directed folate receptor targeting using liposomes is usually achieved using folate linked to a phospholipid or cholesterol anchor. This link is formed using a large spacer like polyethylene glycol. Here, we report an innovative strategy for targeted liposome delivery that uses a hydrophobic fragment of surfactant protein D linked to folate. Our proposed spacer is a small 4 amino acid residue linker. The peptide conjugate inserts deeply into the lipid bilayer without affecting liposomal integrity, with high stability and specificity. To compare the drug delivery potential of both liposomal targeting systems, we encapsulated the nuclear dye Hoechst 34580. The eventual increase in blue fluorescence would only be detectable upon liposome disruption, leading to specific binding of this dye to DNA. Our delivery system was proven to be more efficient (2-fold) in Caco-2 cells than classic systems where the folate moiety is linked to liposomes by polyethylene glycol. PMID:26241560

  3. Considerations on the design of through-wall anchors

    Connections to existing buildings are often the most difficult planning challenge for the realization of construction measures in case of piping system replacements in nuclear power plants. This is due to restricted space or limited load reserves of the building structure. Usually the realization of support connections to the existing buildings is achieved by anchor bolts. But in critical cases the preferred alternative solution uses through-wall anchors. Up to now uniform assessment thresholds are not available, no technical guidelines or regulations for construction variants exist. Through-wall anchors allow significantly higher load capacities for tensile and shear loads but require enhanced planning and realization efforts.

  4. Neurostimulation leads, intrathecal catheters and anchoring devices evolution

    Laura Demartini

    2015-03-01

    Full Text Available Many scientific studies highlight the usefulness of spinal cord stimulation and intrathecal therapy for the management of chronic pain syndromes resistant to pharmacological or less invasive interventional therapies. One of the possible complications of these techniques, reported in literature, is migration of the lead or catheter; thus the use of an anchoring system is considered mandatory. Every company that produces devices for neurostimulation or neuromodulation provides various anchoring devices evolved over time. In the study, the authors discuss about the most common anchoring devices based on their clinical experience.

  5. Maxillary sinus perforation by orthodontic anchor screws.

    Motoyoshi, Mitsuru; Sanuki-Suzuki, Rina; Uchida, Yasuki; Saiki, Akari; Shimizu, Noriyoshi

    2015-06-01

    To facilitate safe placement of orthodontic anchor screws (miniscrews), we investigated the frequency of maxillary sinus perforation after screw placement and the effect of sinus perforation on screw stability. Maxillary sinus perforations involving 82 miniscrews (diameter, 1.6 mm; length, 8 mm) were evaluated using cone-beam computed tomography. All miniscrews were placed in maxillary alveolar bone between the second premolar and first molar for anchorage for anterior retraction in patients undergoing first premolar extraction. The placement torque and screw mobility of each implant were determined using a torque tester and a Periotest device, and variability in these values in relation to sinus perforation was evaluated. Eight of the 82 miniscrews perforated the maxillary sinus. There was no case of sinusitis in patients with miniscrew perforation and no significant difference in screw mobility or placement torque between perforating and non-perforating miniscrews. The sinus floor was significantly thinner in perforated cases than in non-perforated cases. A sinus floor thickness of 6.0 mm or more is recommended in order to avoid miniscrew perforation of the maxillary sinus. PMID:26062857

  6. Anchored boundary conditions for locally isostatic networks

    Theran, Louis; Nixon, Anthony; Ross, Elissa; Sadjadi, Mahdi; Servatius, Brigitte; Thorpe, M. F.

    2015-11-01

    Finite pieces of locally isostatic networks have a large number of floppy modes because of missing constraints at the surface. Here we show that by imposing suitable boundary conditions at the surface the network can be rendered effectively isostatic. We refer to these as anchored boundary conditions. An important example is formed by a two-dimensional network of corner sharing triangles, which is the focus of this paper. Another way of rendering such networks isostatic is by adding an external wire along which all unpinned vertices can slide (sliding boundary conditions). This approach also allows for the incorporation of boundaries associated with internal holes and complex sample geometries, which are illustrated with examples. The recent synthesis of bilayers of vitreous silica has provided impetus for this work. Experimental results from the imaging of finite pieces at the atomic level need such boundary conditions, if the observed structure is to be computer refined so that the interior atoms have the perception of being in an infinite isostatic environment.

  7. High-resolution structure of TBP with TAF1 reveals anchoring patterns in transcriptional regulation

    Anandapadamanaban, Madhanagopal; Andresen, Cecilia; Helander, Sara; Ohyama, Yoshifumi; Siponen, Marina I.; Lundström, Patrik; Kokubo, Tetsuro; Ikura, Mitsuhiko; Moche, Martin; Sunnerhagen, Maria

    2016-01-01

    The general transcription factor TFIID provides a regulatory platform for transcription initiation. Here we present the crystal structure (1.97 Å) and NMR analysis of yeast TAF1 N-terminal domains TAND1 and TAND2 when bound to yeast TBP, together with mutational data. The yTAF1-TAND1, which in itself acts as a transcriptional activator, binds into the DNA-binding TBP concave surface by presenting similar anchor residues to TBP as E. coli Mot1 but from a distinct structural scaffold. Furthermore, we show how yTAF1-TAND2 employs an aromatic and acidic anchoring pattern to bind a conserved yTBP surface groove traversing the basic helix region, and we find highly similar TBP-binding motifs also presented by the structurally distinct TFIIA, Mot1 and Brf1 proteins. Our identification of these anchoring patterns, which can be easily disrupted or enhanced, provides compelling insight into the competitive multiprotein TBP interplay critical to transcriptional regulation. PMID:23851461

  8. Stone anchors from the Okhamandal region, Gujarat Coast, India

    Sundaresh; Gaur, A.S.; Gudigar, P.; Tripati, S.; Vora, K.H.; Bandodkar, S.N.

    Dwarka and Armada yielded a number of stone anchors which suggest that Okhamandal area was major concentration of maritime activities in Saurashtra region in the past. Among these three sites, Bet Dwarka has clear evidence on the settlement of Harappan...

  9. The Carboxy-Terminal Sequence of the Pestivirus Glycoprotein Erns Represents an Unusual Type of Membrane Anchor

    Fetzer, Christiane; Tews, Birke Andrea; Meyers, Gregor

    2005-01-01

    The Erns protein is a structural glycoprotein of pestiviruses that lacks a typical membrane anchor sequence and is known to be secreted from the infected cell. However, major amounts of the protein are retained within the cell and attached to the virion by a so far unknown mechanism. Transient-expression studies with cDNA constructs showed that in a steady-state situation, 16% of the protein is found in the supernatant of the transfected cells while 84% appears as intracellular protein. We sh...

  10. Lentiviral Engineered Fibroblasts Expressing Codon-Optimized COL7A1 Restore Anchoring Fibrils in RDEB

    Georgiadis, Christos; Syed, Farhatullah; Petrova, Anastasia; Abdul-Wahab, Alya; Lwin, Su M.; Farzaneh, Farzin; Chan, Lucas; Ghani, Sumera; Fleck, Roland A.; Glover, Leanne; McMillan, James R.; Chen, Mei; Thrasher, Adrian J.; McGrath, John A.; Di, Wei-Li; Qasim, Waseem

    2016-01-01

    Cells therapies, engineered to secrete replacement proteins, are being developed to ameliorate otherwise debilitating diseases. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects of type VII collagen, a protein essential for anchoring fibril formation at the dermal-epidermal junction. Whereas allogeneic fibroblasts injected directly into the dermis can mediate transient disease modulation, autologous gene-modified fibroblasts should evade immunological rejection and support sustained delivery of type VII collagen at the dermal-epidermal junction. We demonstrate the feasibility of such an approach using a therapeutic grade, self-inactivating-lentiviral vector, encoding codon-optimized COL7A1, to transduce RDEB fibroblasts under conditions suitable for clinical application. Expression and secretion of type VII collagen was confirmed with transduced cells exhibiting supranormal levels of protein expression, and ex vivo migration of fibroblasts was restored in functional assays. Gene-modified RDEB fibroblasts also deposited type VII collagen at the dermal-epidermal junction of human RDEB skin xenografts placed on NOD-scid IL2Rgammanull recipients, with reconstruction of human epidermal structure and regeneration of anchoring fibrils at the dermal-epidermal junction. Fibroblast-mediated restoration of protein and structural defects in this RDEB model strongly supports proposed therapeutic applications in man. PMID:26763448

  11. Neurostimulation leads, intrathecal catheters and anchoring devices evolution

    Laura Demartini; Gianluca Conversa; Luciana Armiento; Lucia Angelini; Cesare Bonezzi

    2015-01-01

    Many scientific studies highlight the usefulness of spinal cord stimulation and intrathecal therapy for the management of chronic pain syndromes resistant to pharmacological or less invasive interventional therapies. One of the possible complications of these techniques, reported in literature, is migration of the lead or catheter; thus the use of an anchoring system is considered mandatory. Every company that produces devices for neurostimulation or neuromodulation provides various anchoring...

  12. Monetary sterilization and dual nominal anchors: some Caribbean examples

    Khemraj, Tarron; Pasha, Sukrishnalall

    2011-01-01

    This paper notes that a high sterilization coefficient plus a de facto pegged exchange rate indicates the existence of dual nominal anchors. The econometric evidence presented shows that several Caribbean economies with fixed exchange rate regimes also possess high sterilization coefficients. Given open capital accounts in the various economies, the paper argues that this finding contravenes the money neutrality thesis, which holds that only one nominal anchor can prevail in the long-term. Th...

  13. Epidermolysis Bullosa Acquisita: Autoimmunity to Anchoring Fibril Collagen

    Chen, Mei; Kim, Gene H; Prakash, Lori; Woodley, David T.

    2011-01-01

    Epidermolysis bullosa acquisita (EBA) is a rare and acquired autoimmune subepidermal bullous disease of the skin and mucosa. EBA includes various distinct clinical manifestations resembling Bullous Pemphigus, Brunsting-Perry pemphigoid, or cicatricial pemphigoid. These patients have autoantibodies against type VII collagen, an integral component of anchoring fibrils, which are responsible for attaching the dermis to the epidermis. Destruction or perturbation of the normally functioning anchor...

  14. Liquid crystal director fluctuations and surface anchoring by molecular simulation

    Andrienko, D.; Germano, G; Allen, M. P.

    2002-01-01

    We propose a simple and reliable method to measure the liquid crystal surface anchoring strength by molecular simulation. The method is based on the measurement of the long-range fluctuation modes of the director in confined geometry. As an example, molecular simulations of a liquid crystal in slab geometry between parallel walls with homeotropic anchoring have been carried out using the Monte Carlo technique. By studying different slab thicknesses, we are able to calculate separately the pos...

  15. Transosseous Acetabular Labral Repair as an Alternative to Anchors

    Pérez-Carro, Luis; Cabello, Andres Gonzalez; Rakha, Mohamed Ibrahim; Patnaik, Sarthak; Centeno, Elias; Miranda, Victor; Fernández, Ana Alfonso

    2015-01-01

    Labral tears are the most common pathology in patients undergoing hip arthroscopy and the most common cause of mechanical hip symptoms. Labral repair techniques have been described in the literature using suture anchors placed as close as possible to the acetabular rim without penetrating the articular surface. Optimal surgical technique for labral repair is very important, and an inappropriate entry point and guide angulation may lead to intra-articular penetration of the anchor, chondral da...

  16. The Effect of Anchoring Sutures on Medicinal Leech Mortality

    Davila, Victor J.; Hoppe, Ian C.; Landi, Rocco; Ciminello, Frank S.

    2009-01-01

    Objective: The implementation of leech therapy for surgical flaps is not always logistically easy or comfortable for patients or healthcare providers. We examine different methods of placing sutures in the medicinal leech, Hirudo medicinalis, to make the implementation of leech therapy easier. Methods: Sixteen leeches were randomly divided into 3 groups: a control group, a deep anchoring suture group, and a superficial anchoring suture group. The leeches were observed to determine if either o...

  17. Anchor Plates in Two-Layered Cohesion Less Soils

    Hamed Niroumand

    2010-01-01

    Full Text Available Problem statement: During the past few years a great number of experimental model and numerical analysis results on the uplift resistance of anchor plate embedded in homogeneous cohesion less soil has been reported by many researchers. A review of related literature shows that not much research has been done to analyze the performance of anchor plates in layered soils a problem, which is often encountered by the professional engineers in the field. Approach: This study presented the performance of the anchor plates in the cohesion less soil by different researchers. It was based on different previous researches, from the earliest till the most recent ones. The main aim of this research was focused on the prediction of the anchor plates behavior and the force in the layered cohesion less soils. Few laboratory studies were conducted to investigate the uplift capacity behavior of cohesion less soil by previous researchers. Results: The experimental and numerical investigation included uplift test on cohesion less soil by last researchers. The embedment ratio and the effect of density on uplift response were evaluated. This analysis was investigated experimentally and numerically behavior of anchor plates buried in two layered cohesion less soil. Although earlier researchers developed experimentally expressions to estimate the uplift capacity of irregular anchor plates in layered cohesion less soils. Conclusion: The study observed that the ultimate uplift capacity is dependent on the relative strength of the two layers, the depth ratio of embedment and the upper layer thickness ratio.

  18. Anchor stud monotonic and cyclic shear tests. Final report

    In a prestressed concrete reactor vessel (PCRV), the main cavity liner and other cavity liners with large diameter/thickness (D/t) ratios are anchored to the concrete by anchor studs. These anchor studs are subject to shear loading resulting from prestressing, pressurization, temperature, and creep of the PCRV. The test program discussed in this report consists of both monotonic and cyclic testing of models simulating the prototype anchor stud/concrete assembly. The tests determined the shear stiffness characteristics of the 3/4-in.-diameter, one-piece Nelson anchor stud embedded in concrete and established its low-cycle fatigue life under displacement-controlled loading. The previously obtained results of Phase I and Phase II tests on two-piece anchor studs, along with high-cyclic fatigue data taken from the literature, are discussed and are plotted with the current, Phase III, results to form a displacement versus cycles to failure curve covering a useful range of design applications

  19. Evaluation of mitral valve replacement anchoring in a phantom

    McLeod, A. Jonathan; Moore, John; Lang, Pencilla; Bainbridge, Dan; Campbell, Gordon; Jones, Doug L.; Guiraudon, Gerard M.; Peters, Terry M.

    2012-02-01

    Conventional mitral valve replacement requires a median sternotomy and cardio-pulmonary bypass with aortic crossclamping and is associated with significant mortality and morbidity which could be reduced by performing the procedure off-pump. Replacing the mitral valve in the closed, off-pump, beating heart requires extensive development and validation of surgical and imaging techniques. Image guidance systems and surgical access for off-pump mitral valve replacement have been previously developed, allowing the prosthetic valve to be safely introduced into the left atrium and inserted into the mitral annulus. The major remaining challenge is to design a method of securely anchoring the prosthetic valve inside the beating heart. The development of anchoring techniques has been hampered by the expense and difficulty in conducting large animal studies. In this paper, we demonstrate how prosthetic valve anchoring may be evaluated in a dynamic phantom. The phantom provides a consistent testing environment where pressure measurements and Doppler ultrasound can be used to monitor and assess the valve anchoring procedures, detecting pararvalvular leak when valve anchoring is inadequate. Minimally invasive anchoring techniques may be directly compared to the current gold standard of valves sutured under direct vision, providing a useful tool for the validation of new surgical instruments.

  20. Pyramidal anchor stone from Baga waters of Goa, west coast of India

    Tripati, S.

    . Pyramidal anchor stones have an apex hole which goes up to the round hole, however Goa anchor stone has no such perforation, but, instead has a rectangular cutting on the apex. The anchor stone is compared with Greek pyramidal anchor stones, and probably...

  1. Glycolipid precursors for the membrane anchor of Trypanosoma brucei variant surface glycoproteins. II. Lipid structures of phosphatidylinositol-specific phospholipase C sensitive and resistant glycolipids

    A common diagnostic feature of glycosylinositol phospholipid (GPI)-anchored proteins is their release from the membrane by a phosphatidylinositol-specific phospholipase C (PI-PLC). However, some GPI-anchored proteins are resistant to this enzyme. The best characterized example of this subclass is the human erythrocyte acetylcholinesterase, where the structural basis of PI-PLC resistance has been shown to be the acylation of an inositol hydroxyl group(s). Both PI-PLC-sensitive and resistant GPI-anchor precursors (P2 and P3, respectively) have been found in Trypanosoma brucei, where the major surface glycoprotein is anchored by a PI-PLC-sensitive glycolipid anchor. The accompanying paper shows that P2 and P3 have identical glycans, indistinguishable from the common core glycan found on all the characterized GPI protein anchors. This paper shows that the single difference between P2 and P3, and the basis for the PI-PLC insusceptibility of P3, is a fatty acid, ester-linked to the inositol residue in P3. The inositol-linked fatty acid can be removed by treatment with mild base to restore PI-PLC sensitivity. Biosynthetic labeling experiments with [3H]palmitic acid and [3H]myristic acid show that [3H]palmitic acid specifically labels the inositol residue in P3 while [3H]myristic acid labels the diacylglycerol portion. Possible models to account for the simultaneous presence of PI-PLC-resistant and sensitive glycolipids are discussed in the context of available information on the biosynthesis of GPI-anchors

  2. Support Vector Machines for Predicting Membrane Protein Types by Using Functional Domain Composition

    Cai, Yu-Dong; Zhou, Guo-Ping; Chou, Kuo-Chen

    2003-01-01

    Membrane proteins are generally classified into the following five types: 1), type I membrane protein; 2), type II membrane protein; 3), multipass transmembrane proteins; 4), lipid chain-anchored membrane proteins; and 5), GPI-anchored membrane proteins. In this article, based on the concept of using the functional domain composition to define a protein, the Support Vector Machine algorithm is developed for predicting the membrane protein type. High success rates are obtained by both the self...

  3. Display of GPI-anchored anti-EGFR nanobodies on extracellular vesicles promotes tumour cell targeting

    Sander A. A. Kooijmans

    2016-03-01

    Full Text Available Background: Extracellular vesicles (EVs are attractive candidate drug delivery systems due to their ability to functionally transport biological cargo to recipient cells. However, the apparent lack of target cell specificity of exogenously administered EVs limits their therapeutic applicability. In this study, we propose a novel method to equip EVs with targeting properties, in order to improve their interaction with tumour cells. Methods: EV producing cells were transfected with vectors encoding for anti-epidermal growth factor receptor (EGFR nanobodies, which served as targeting ligands for tumour cells, fused to glycosylphosphatidylinositol (GPI anchor signal peptides derived from decay-accelerating factor (DAF. EVs were isolated using ultrafiltration/size-exclusion liquid chromatography and characterized using western blotting, Nanoparticle Tracking Analysis, and electron microscopy. EV–tumour cell interactions were analyzed under static conditions using flow cytometry and under flow conditions using a live-cell fluorescence microscopy-coupled perfusion system. Results: V analysis showed that GPI-linked nanobodies were successfully displayed on EV surfaces and were highly enriched in EVs compared with parent cells. Display of GPI-linked nanobodies on EVs did not alter general EV characteristics (i.e. morphology, size distribution and protein marker expression, but greatly improved EV binding to tumour cells dependent on EGFR density under static conditions. Moreover, nanobody-displaying EVs showed a significantly improved cell association to EGFR-expressing tumour cells under flow conditions. Conclusions: We show that nanobodies can be anchored on the surface of EVs via GPI, which alters their cell targeting behaviour. Furthermore, this study highlights GPI-anchoring as a new tool in the EV toolbox, which may be applied for EV display of a variety of proteins, such as antibodies, reporter proteins and signaling molecules.

  4. Cell wall sorting signals in surface proteins of gram-positive bacteria.

    Schneewind, O; Mihaylova-Petkov, D; Model, P

    1993-01-01

    Staphylococcal protein A is anchored to the cell wall, a unique cellular compartment of Gram-positive bacteria. The sorting signal sufficient for cell wall anchoring consists of an LPXTG motif, a C-terminal hydrophobic domain and a charged tail. Homologous sequences are found in many surface proteins of Gram-positive bacteria and we explored the universality of these sequences to serve as cell wall sorting signals. We show that several signals are able to anchor fusion proteins to the staphyl...

  5. Perceptual anchoring in preschool children: not adultlike, but there.

    Karen Banai

    Full Text Available BACKGROUND: Recent studies suggest that human auditory perception follows a prolonged developmental trajectory, sometimes continuing well into adolescence. Whereas both sensory and cognitive accounts have been proposed, the development of the ability to base current perceptual decisions on prior information, an ability that strongly benefits adult perception, has not been directly explored. Here we ask whether the auditory frequency discrimination of preschool children also improves when given the opportunity to use previously presented standard stimuli as perceptual anchors, and whether the magnitude of this anchoring effect undergoes developmental changes. METHODOLOGY/PRINCIPAL FINDINGS: Frequency discrimination was tested using two adaptive same/different protocols. In one protocol (with-reference, a repeated 1-kHz standard tone was presented repeatedly across trials. In the other (no-reference, no such repetitions occurred. Verbal memory and early reading skills were also evaluated to determine if the pattern of correlations between frequency discrimination, memory and literacy is similar to that previously reported in older children and adults. Preschool children were significantly more sensitive in the with-reference than in the no-reference condition, but the magnitude of this anchoring effect was smaller than that observed in adults. The pattern of correlations among discrimination thresholds, memory and literacy replicated previous reports in older children. CONCLUSIONS/SIGNIFICANCE: The processes allowing the use of context to form perceptual anchors are already functional among preschool children, albeit to a lesser extent than in adults. Nevertheless, immature anchoring cannot fully account for the poorer frequency discrimination abilities of young children. That anchoring is present among the majority of typically developing preschool children suggests that the anchoring deficits observed among individuals with dyslexia represent a

  6. Protein complex analysis of native brain potassium channels by proteomics.

    Sandoz, Guillaume; Lesage, Florian

    2008-01-01

    TREK potassium channels belong to a family of channel subunits with two-pore domains (K(2P)). TREK1 knockout mice display impaired polyunsaturated fatty acid-mediated protection against brain ischemia, reduced sensitivity to volatile anesthetics, resistance to depression and altered perception of pain. Recently, we isolated native TREK1 channels from mouse brain and identified their specific components by mass spectrometry. Among the identified partners, the A-Kinase Anchoring Protein AKAP150 binds to a regulatory domain of TREK1 and acts as a molecular switch. It transforms low activity, outwardly rectifying TREK1 currents into robust leak conductances resistant to stimulation by arachidonic acid, membrane stretch and acidification. Inhibition of the TREK1/AKAP150 channel by Gs-coupled receptors is as extensive as for TREK1 alone (but faster) whereas inhibition of TREK1/AKAP150 by Gq-coupled receptors is reduced. Furthermore, the association of AKAP150 with TREK1 channels integrates them into postsynaptic scaffolds where G protein-coupled membrane receptors and channels dock simultaneously. This chapter describes the proteomic approach used to study the composition of native TREK1 channels and point out its advantages and limitations over more classical methods (two-hybrid screenings in the yeast and bacteria or GST-pull down). PMID:18998088

  7. Effects of Anchor Bolts Failures in Steam Explosion Analyses

    Kim, Seung Hyun; Chang, Yoon-Suk [Kyung Hee University, Yongin (Korea, Republic of); Song, Sungchu; Cho, Yong-Jin [Korea Institute of Nuclear Safety, Daejeon (Korea, Republic of)

    2015-05-15

    Steam explosion may occur in a nuclear power plant by molten core-coolant interactions when the external reactor vessel cooling strategy is failed. This phenomenon can threat the integrity of reactor cavity, penetration piping and support structures. Even though extensive researches have been performed to predict influences of the steam explosion, due to complexity of physical phenomena and environmental thermal hydraulic conditions, it is remained as one of possible hazards. A steam explosion can cause intensive and rapid heat transfer, and lead to the formation of pressure waves and production of missiles that may endanger surrounding reactor cavity wall and associate components due to resulting dynamic effects. The goal of this research is to examine structural integrity of RPV (Reactor Pressure Vessel) support structures and anchor bolts under typical ex-vessel steam explosion conditions through FE analyses. Particularly, influence due to the failure of anchor bolts connecting RPV and support structures was evaluated. In this paper, influence of RPV and support structure due to the anchor bolt failure were evaluated under typical steam explosion conditions and the following conclusions were derived. The highest maximum stresses were calculated at the support structures under the steam explosion condition with the SVF and anchor bolts non-failure. The all stress values did not exceed their yield strengths. The displacements were high under anchor bolt failure conditions. However, the vertical movements of major components were small comparing to the overall dimensions of them.

  8. Clustering of sialylated glycosylphosphatidylinositol anchors mediates PrP-induced activation of cytoplasmic phospholipase A2 and synapse damage

    Bate, Clive; Williams, Alun

    2012-01-01

    Precisely how the accumulation of PrPSc causes the neuronal degeneration that leads to the clinical symptoms of prion diseases is poorly understood. Our recent paper showed that the clustering of specific glycosylphosphatidylinositol (GPI) anchors attached to PrP proteins triggered synapse damage in cultured neurons. First, we demonstrated that small, soluble PrPSc oligomers caused synapse damage via a GPI-dependent process. Our hypothesis, that the clustering of specific GPIs caused synapse ...

  9. Chimeric Rabies Virus-Like Particles Containing Membrane-Anchored GM-CSF Enhances the Immune Response against Rabies Virus

    Hongtao Kang; Yinglin Qi; Hualei Wang; Xuexing Zheng; Yuwei Gao; Nan Li; Songtao Yang; Xianzhu Xia

    2015-01-01

    Rabies remains an important public health threat in most developing countries. To develop a more effective and safe vaccine against rabies, we have constructed a chimeric rabies virus-like particle (VLP), which containing glycoprotein (G) and matrix protein (M) of rabies virus (RABV) Evelyn-Rokitnicki-Abelseth (ERA) strain, and membrane-anchored granulocyte-macrophage colony-stimulating factor (GM-CSF), and it was named of EVLP-G. The immunogenicity and protective efficacy of EVLP-G against R...

  10. Sortase A substrate specificity in GBS pilus 2a cell wall anchoring.

    Francesca Necchi

    Full Text Available Streptococcus agalactiae, also referred to as Group B Streptococcus (GBS, is one of the most common causes of life-threatening bacterial infections in infants. In recent years cell surface pili have been identified in several Gram-positive bacteria, including GBS, as important virulence factors and promising vaccine candidates. In GBS, three structurally distinct types of pili have been discovered (pilus 1, 2a and 2b, whose structural subunits are assembled in high-molecular weight polymers by specific class C sortases. In addition, the highly conserved housekeeping sortase A (SrtA, whose main role is to link surface proteins to bacterial cell wall peptidoglycan by a transpeptidation reaction, is also involved in pili cell wall anchoring in many bacteria. Through in vivo mutagenesis, we demonstrate that the LPXTG sorting signal of the minor ancillary protein (AP2 is essential for pilus 2a anchoring. We successfully produced a highly purified recombinant SrtA (SrtA(ΔN40 able to specifically hydrolyze the sorting signal of pilus 2a minor ancillary protein (AP2-2a and catalyze in vitro the transpeptidation reaction between peptidoglycan analogues and the LPXTG motif, using both synthetic fluorescent peptides and recombinant proteins. By contrast, SrtA(ΔN40 does not catalyze the transpeptidation reaction with substrate-peptides mimicking sorting signals of the other pilus 2a subunits (the backbone protein and the major ancillary protein. Thus, our results add further insight into the proposed model of GBS pilus 2a assembly, in which SrtA is required for pili cell wall covalent attachment, acting exclusively on the minor accessory pilin, representing the terminal subunit located at the base of the pilus.

  11. Plate anchored stirrup; Pureto teichakugata sendan hokyo tekkin

    Nakamura, T.; Tanaka, Y.; Nakano, K.; Yamamoto, T. [Taisei Construction Co. Ltd., Tokyo (Japan)

    1998-09-01

    After the Great Hanshin Earthquake, the use of edge form of stirrups of semicircular hook or acute-angled hook which functions sufficiently even after exfoliation of concrete cover is proposed instead of right angle hook. However, when these stirrups are used in plate structures requiring hooks on two ends or in columns as connection reinforcement of hoop, the construction of these stirrups is so complicated that brings higher cost. The authors proposes stirrups to anchor by plate for stirrups of planar members. Anchorage capacity of these plate anchored stirrups and hooks after exfoliation of concrete cover was examined by experiments and the anchorage capacity was confirmed to be larger than that of semicircular hooks. The plate anchored stirrups are effective and easy to construct with low cost. 7 refs., 13 figs., 4 tabs.

  12. Optical Fiber Grating Sensor for Force Measurement of Anchor Cable

    JIANG Desheng; FU Jinghua; LIU Shengchun; SUI Lingfeng; FU Rong

    2006-01-01

    The development of the sensor suitable for measuring large load stress to the anchor cable becomes an important task in bridge construction and maintenance. Therefore, a new type of optical fiber sensor was developed in the laboratory - optical fiber grating sensor for force measurement of anchor cable (OFBFMAC). No similar report about this kind of sensor has been found up to now in China and other countries. This sensor is proved to be an effective way of monitoring in processes of anchor cable installation, cable cutting, cable force regulation, etc, with the accurate and repeatable measuring results. Its successful application in the tie bar cable force safety monitoring for Wuhan Qingchuan bridge is a new exploration of optical fiber grating sensing technology in bridge tie bar monitoring system.

  13. PULLOUT CAPACITY OF HORIZONTAL AND INCLINED PLATE ANCHORS IN CLAYEY SOILS

    BALESHWAR SINGH

    2011-11-01

    Full Text Available Soil anchors are required to withstand uplift or lateral forces acting on the foundations of structures constructed both in land and offshore sites. Plate anchoring system is being increasingly used to moor floating structures for the exploration and development of oil and gas fields. In this study, various experimental,theoretical and numerical approaches for estimation of pullout capacity of horizontal and inclined plate anchors in clayey soils are reviewed. A comparative analysis of the ultimate capacity is then carried out for plate anchors embedded in clayey soils by varying embedment ratio for horizontal anchors and by varying inclination angle for inclined plate anchors.

  14. Current status of frameless anchored IUD for immediate intracesarean insertion.

    Wildemeersch, Dirk; Goldstuck, Norman D; Hasskamp, Thomas

    2016-01-01

    Immediate postpartum intrauterine device (IUD) insertion deserves great attention as it can provide immediate, timely and convenient contraception plus the added benefit of preventing repeat unintended pregnancies. Although women post vaginal delivery can benefit from immediate post-placenta contraception, women undergoing Cesarean section clearly need contraception, as an inter-delivery interval shorter than 18 months places them at a high risk for uterine rupture. The main drawback of currently available framed IUD devices for immediate postpartum insertion of an IUD is their high expulsion and displacement rates when inserted immediately postpartum after both vaginal and Cesarean delivery. Current research suggests that a brief window of opportunity exists of 10 minutes for insertion of conventional IUDs after which time expulsion rates both immediately and over time are greatly enhanced. This paper summarizes the current research conducted to overcome the expulsion problems associated with conventional T-shaped devices as well as through the use of an anchored frameless device. In the 1970s and 1980s, attempts were made to solve the expulsion problem by modifying existing devices, such as adding absorbable sutures (Delta-T) or additional appendages. These attempts proved to be clinically unsuccessful as the catgut suture added to the transverse arms did not provide sufficient resistance to prevent downward displacement and expulsion. An anchoring technique to suspend a copper IUD to the fundus of the uterus was developed in Belgium in the 1980s and has been the subject of extensive ongoing clinical research since 1985. Recently the frameless copper releasing anchor IUD, GyneFix, has been tested for postplacental insertion. Initially, the anchor was modified by the inclusion of a biodegradable cone which was added below the anchoring knot. Clinical studies confirmed the adequacy of this approach suggesting that it was technically possible to anchor an IUD

  15. [Dactylogyrids (Monogenea: Dactylogyridea) with an unusual number of anchors, their origin and phylogenetic significance. Reference data].

    Gerasev, P I

    2002-01-01

    It is considered that in Anacanthorinae, Markewitschiana and Pseudacolpenteron pavlovskii (Dactylogyridae s. s.) the anchors are absent originally. In the other dactylogyrids (s. l.) without anchors (Pseudacolpenteron ignotus; Acolpenteron; Anonchohaptor-Icelanchohaptor; Kritskyia-Telethecium-Pavanelliela) these structures are lost. In related genera Rhinoxenus and Nanotrema the dorsal pair of anchors is transformed into the "spikes"; in Heteronchocleidinae there is one anchor; in Trinigyrus and Schilbetrematoides are two anchors; and in Urogyrus three anchors are reduced. But in Rhinonaster, Cacatyocotyle, Callceostomella, Neocalseostoma elongatum and Pseudodactylogyridae the presence of one pair of ventral "peduncular" or "haptoral" anchors appear to be the original state. For Anacanthorinae and Dactylogyridae s. s. the presence of 18-16 hooks, original lacking of anchors, appearance of one pair of them, evolutionary development and subsequent reduction of these structures are characteristic. In the dactylogyrids (s. l.) having two pairs of anchors and 14 hooks, one pair of the anchors is present originally, then usually the second dorsal pair grows up, and finally, the reduction of a part of anchors or all the anchors is possible. In the 16-hooked dactylogyrids (s. l.) having two pairs of anchors (Tetraonchinea, Amphibdellainea, Neodactylodiscinea) the original lacking or reduction of the anchors is not reported. The author considers, that it is unwarranted to unite all dactylogyrids (s. l.) without anchors into the superorder Ananchorea Malmberg, 1990 or to postulate the original presence of the two pairs of ventral anchors in this group. It is also impossible to unit all dactylogyrids (s. l.) having 0-4 anchors and 18-14 hooks into Dactylogyridae sensu Boeger et Kritsky, 1993. PMID:12070963

  16. SNP Discovery and Chromosome Anchoring Provide the First Physically-Anchored Hexaploid Oat Map and Reveal Synteny with Model Species.

    Rebekah E Oliver; Tinker, Nicholas A.; Lazo, Gerard R.; Shiaoman Chao; Jellen, Eric N.; Martin L. Carson; Rines, Howard W; Donald E Obert; Lutz, Joseph D.; Irene Shackelford; Korol, Abraham B.; Charlene P. Wight; Gardner, Kyle M.; Jiro Hattori; Beattie, Aaron D

    2013-01-01

    A physically anchored consensus map is foundational to modern genomics research; however, construction of such a map in oat (Avena sativa L., 2n = 6x = 42) has been hindered by the size and complexity of the genome, the scarcity of robust molecular markers, and the lack of aneuploid stocks. Resources developed in this study include a modified SNP discovery method for complex genomes, a diverse set of oat SNP markers, and a novel chromosome-deficient SNP anchoring strategy. These resources wer...

  17. The Assembly of Proline-rich Membrane Anchor (PRiMA)-linked Acetylcholinesterase Enzyme: GLYCOSYLATION IS REQUIRED FOR ENZYMATIC ACTIVITY BUT NOT FOR OLIGOMERIZATION*

    Chen, Vicky P.; Choi, Roy C. Y.; Chan, Wallace K. B.; Leung, K. Wing; Guo, Ava J. Y.; Gallant K. L. Chan; Luk, Wilson K. W.; Tsim, Karl W. K.

    2011-01-01

    Acetylcholinesterase (AChE) anchors onto cell membranes by a transmembrane protein PRiMA (proline-rich membrane anchor) as a tetrameric form in vertebrate brain. The assembly of AChE tetramer with PRiMA requires the C-terminal “t-peptide” in AChE catalytic subunit (AChET). Although mature AChE is well known N-glycosylated, the role of glycosylation in forming the physiologically active PRiMA-linked AChE tetramer has not been studied. Here, several lines of evidence indicate that the N-linked ...

  18. Characterization of the phosphatidylinositol-glycan membrane anchor of human placental alkaline phosphatase

    Placental alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] is a member of a diverse group of membrane proteins whose attachment to the lipid bilayer is mediated by a phosphatidylinositol-glycan. To investigate structural aspects of the glycolipid anchor, cultured WISH cells were used because, they produce the enzyme in abundant quantities. When cell suspensions were incubated with purified phosphatidylinositol-specific phospholipase C, most of the placental alkaline phosphatase was released from membranes in a hydrophilic form. On incubation of the cells with [14C]ethanolamine, [14C]myristic acid, or myo[3H]inositol, each was incorporated into the phosphatase near the carboxyl terminus, showing that these components, which are found in other phosphatidylinositol membrane-linked proteins, are also present in placental alkaline phosphatase

  19. Localization in Wireless Sensor Networks and Anchor Placement

    Benjamin Tatham

    2012-06-01

    Full Text Available Applications of wireless sensor network (WSN often expect knowledge of the precise location of the nodes. Many different localization protocols have been proposed that allow nodes to derive their location rather than equipping them with dedicated localization hardware such as GPS receivers, which increases node costs. We provide a brief survey of the major approaches to software-based node localization in WSN. One class of localization protocols with good localization performance patches together relative-coordinate, local maps into a global-coordinate map. These protocols require some nodes that know their absolute coordinates, called anchor nodes. While many factors influence the node position errors, in this class of protocols, using Procrustes Analysis, the placement of the anchor nodes can significantly impact the error. Through simulation, using the Curvilinear Component Analysis (CCA-MAP protocol as a representative protocol in this category, we show the impact of anchor node placement and propose a set of guidelines to ensure the best possible outcome, while using the smallest number of anchor nodes possible.

  20. When Does Scale Anchoring Work? A Case Study

    Sinharay, Sandip; Haberman, Shelby J.; Lee, Yi-Hsuan

    2011-01-01

    Providing information to test takers and test score users about the abilities of test takers at different score levels has been a persistent problem in educational and psychological measurement. Scale anchoring, a technique which describes what students at different points on a score scale know and can do, is a tool to provide such information.…

  1. The importance of anchor text for ad hoc search revisited

    M. Koolen; J. Kamps

    2010-01-01

    It is generally believed that propagated anchor text is very important for effective Web search as offered by the commercial search engines. "Google Bombs" are a notable illustration of this. However, many years of TREC Web retrieval research failed to establish the effectiveness of link evidence fo

  2. Impact of Enhanced Anchored Instruction in Inclusive Math Classrooms

    Bottge, Brian A.; Toland, Michael D.; Gassaway, Linda; Butler, Mark; Choo, Sam; Griffen, Ann Katherine; Ma, Xin

    2015-01-01

    The Common Core State Standards for Mathematics will place more pressure on special education and math teachers to raise the skill levels of all students, especially those with disabilities in math (MD). The purpose of this study was to assess the effects of enhanced anchored instruction (EAI) on students with and without MD in co-taught general…

  3. 76 FR 10627 - Assumption Buster Workshop: Trust Anchors Are Invulnerable

    2011-02-25

    ...) Committee, an interagency working group that coordinates cyber security research activities in support of... confident that they are immune from the attacks.'' This assertion underlies significant cyber security research and development that is aimed at developing and implementing invulnerable trust anchors,...

  4. Analytical Model for Hook Anchor Pull-Out

    Brincker, Rune; Ulfkjær, J. P.; Adamsen, P.;

    1995-01-01

    A simple analytical model for the pull-out of a hook anchor is presented. The model is based on a simplified version of the fictitious crack model. It is assume that the fracture process is the pull-off of a cone shaped concrete part, simplifying the problem by assuming pure rigid body motions...

  5. Analytical Model for Hook Anchor Pull-out

    Brincker, Rune; Ulfkjær, J. P.; Adamsen, P.;

    A simple analytical model for the pull-out of a hook anchor is presented. The model is based on a simplified version of the fictitious crack model. It is assumed that the fracture process is the pull-off of a cone shaped concrete part, simplifying the problem by assuming pure rigid body motions...

  6. Fullerene-based Anchoring Groups for Molecular Electronics

    Martin, Christian A.; Ding, Dapeng; Sørensen, Jakob Kryger;

    2008-01-01

    We present results on a new fullerene-based anchoring group for molecular electronics. Using lithographic mechanically controllable break junctions in vacuum we have determined the conductance and stability of single-molecule junctions of 1,4-bis(fullero[c]pyrrolidin-1-yl)benzene. The compound can...

  7. Experience-based, body-anchored qualitative research interviewing

    Stelter, Reinhard

    2010-01-01

    -anchored interviewing, and second, by an interview guide that explores a research participant's personal experience with mindfulness meditation. An excerpt from an interview is discussed to illustrate the advantages of this interview form, namely its value as a methodological instrument for qualitative research in...

  8. Ten Anchor Points for Teaching Principles of Marketing

    Tomkovick, Chuck

    2004-01-01

    Effective marketing instructors commonly share a love for their students, an affinity for the subject matter, and a devotion to continuous quality improvement. The purpose of this article is to highlight 10 anchor points for teaching Principles of Marketing, which are designed to better engage students in the learning process. These anchor…

  9. A+: Adjusted anchored neighborhood regression for fast super-resolution

    Timofte, Radu; De Smet, Vincent; Van Gool, Luc

    2015-01-01

    Timofte R., De Smet V., Van Gool L., ''A+: Adjusted anchored neighborhood regression for fast super-resolution'', Lecture notes in computer science, vol. 9006, part IV, pp. 111-126, 2015 (12th Asian conference on computer vision - ACCV 2014, November 1-5, 2014, Singapore).

  10. A Single-hole stone anchor from Kottapatnam: Early historic port site of Andhra Pradesh, India

    Tripati, S.; Rao, K.P.; Kumari, S.; Imsong, O.; Vanlalhruaitluangi, V.

    broken piece of polished stone celt in addition to other findings. The recent explorations brought to light a Vijayanagar period coin, pottery and a single hole stone anchor. The finding of the stone anchor throws light on the maritime activities...

  11. Characterizing the flow of stirred vessels with anchor type impellers

    Peixoto S.M.C.

    2000-01-01

    Full Text Available Despite its importance in chemical industries, there are few works which studies anchor type impellers and only a fraction of the works investigate these systems under a computational approach. The great majority refers to turbine impellers, specially Rushton turbines, under turbulent flow. Anchor impellers are used specially for highly viscous flow, typical of polymer reactions. The viscosity is normally in the range 1000-10000 cp. Since this range of viscosity describe highly viscous flows, the reactions for anchor agitated systems are normally carried out under laminar flow. This work presents a detailed computational fluid dynamics (CFD approach to study the behaviour of stirred vessels using anchor impellers. The axial plane of the tank, which is being modelled, is divided into small control volumes, which collectively is referred to as the mesh, or grid. In each of these cells the momentum balance, energy and mass conservation, which describes the model, are rewritten algebraically using the finite volumes method to relate such variables as velocity, pressure and temperature to values in neighbouring cells. The equations are then solved numerically, and the results yield the flow corresponding to the model. Since the geometry of a vessel with anchor impellers strictly calls for a three dimensional method, an approximation is made to account for the effect of the blades (Kuncewics, 1992. The main objective of this work is to give a detailed description of the flow generated by this axial impeller with a view to indicate ways in which the design and operation of these systems can be improved.

  12. Membrane anchoring subunits specify selective regulation of RGS9·Gβ5 GAP complex in photoreceptor neurons

    Cao, Yan; Kolesnikov, Alexander V.; Masuho, Ikuo; Kefalov, Vladimir J.; Martemyanov, Kirill A.

    2010-01-01

    The RGS9·Gβ5 complex is the key regulator of neuronal G protein signaling that shows remarkable selectivity of subunit composition. In retinal photoreceptors, RGS9·Gβ5 is bound to the membrane anchor R9AP and the complex regulates visual signaling. In the basal ganglia neurons, RGS9·Gβ5 is instead associated with a homologous protein, R7BP, and regulates reward circuit. Switching this selective subunit composition of the complex in rod photoreceptors allowed us to study the molecular underpin...

  13. Cement grouting during installation of ground anchors in non-cohesive soils

    Domes, Xenia A. L.

    2015-01-01

    Pressure grouting during installation of grouted ground anchors is known to increase anchor capacity in non-cohesive soils, but little information is available on correlations between applied grouting pressures, duration of grouting, ground conditions and increase of anchor pull-out capacity. The presented PhD study is concerned with processes taking place during installation of grouted ground anchors in non-cohesive soils, where filtration of the cement grout is assumed. It...

  14. An Investigation to Determine the Career Anchors Which Affect the Career Decision of Tourism Students

    GEZEN, Tuba; Köroğlu, Özlem

    2014-01-01

    AbstractAlthough career anchors are used as a career tool in organizations, are examined in many limited researches. The purpose of this study is to determine the career anchors which affect the job and career decision of the tourism students. Therefore, Tourism and Hotel Management students who attend four-year programs/departments of Balikesir University have been conducted surveys. The Career Anchors Scale developed by Schein (1990) has been used to determine the career anchors of the tour...

  15. Developments, expectations of wave energy converters and mooring anchors in the UK

    Huang, Ming; Aggidis, G. A.

    2008-02-01

    The paper introduces the important developments of Wave Energy Converters (WECs) in the UK, and the generic anchor types for WECs and similar structures. Several WECs and their characteristics are introduced to explain the development direction. The anchors are discussed in relation to the behaviour and performance of WECs, and comparisons are made with similar aspects of the offshore industry. Typical and desirable features of anchors for WECs are summarized. Additionally, expectations and research suggestions for WECs and their anchor design are presented.

  16. A group of 20 stone anchors from the waters of Dwarka, on the Gujarat Coast, India

    Gaur, A; Sundaresh; Tripati, S.; Gudigar, P.; Vora, K.H.; Bandodkar, S.N.

    A large number of stone anchors were discovered in a water depth of 10-14 m off Dwarka during the 1998-99 season. The seabed near the anchors consists of a ledge with an average height of 1 m. Several anchors were found trapped between the rocks...

  17. 33 CFR 401.83 - Reporting position at anchor, wharf, etc.

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Reporting position at anchor, wharf, etc. 401.83 Section 401.83 Navigation and Navigable Waters SAINT LAWRENCE SEAWAY DEVELOPMENT... § 401.83 Reporting position at anchor, wharf, etc. A vessel anchoring in a designated anchorage area,...

  18. Biomimetic Mussel Adhesive Inspired Clickable Anchors Applied to the Functionalization of Fe3O4 Nanoparticles

    Goldmann, Anja S.; Schoedel, Christine; Walther, Andreas; Yuan, Jiayin; Loos, Katja; Mueller, Axel H. E.; Müller, Axel H.E.

    2010-01-01

    The functionalization of magnetite (Fe3O4) nanoparticles with dopamine-derived clickable biomimetic anchors is reported. Herein, an alkyne-modified catechol-derivative is employed as the anchor, as i) the catechol-functional anchor groups possess irreversible covalent binding affinity to Fe3O4 nanop

  19. Multivalent anchored and crosslinked hyperbranched polyglycerol monolayers as antifouling coating for titanium oxide surfaces.

    Wei, Qiang; Krysiak, Stefanie; Achazi, Katharina; Becherer, Tobias; Noeske, Paul-Ludwig Michael; Paulus, Florian; Liebe, Hendrik; Grunwald, Ingo; Dernedde, Jens; Hartwig, Andreas; Hugel, Thorsten; Haag, Rainer

    2014-10-01

    A set of new catecholic monolayer coatings was developed to improve the antifouling performance of TiO2 surfaces. To solve the problem of the weak charge-transfer interaction between a single catechol anchor and TiO2, multiple catechol groups were combined with hyperbranched polyglycerol (hPG) which is a distinct dendritic scaffold that exposes its multivalent anchor groups on the surface. Thus, multivalent catecholic hPGs can be easily prepared for surface modification. The immobilization of the compounds was monitored by quartz crystal microbalance with dissipation monitoring. Surface properties of the coatings were analyzed by water contact angle, X-ray photoelectron spectroscopy, and atomic force microscopy. The antifouling ability and stability were investigated by protein adsorption and cell adhesion. By increasing the number of catechol groups on the hPG scaffold, the stability and surface coverage could be significantly enhanced. Moreover, the inner-layer crosslinking of the coatings by grafting and initiating vinyl groups clearly improved their long-term stability. As a result, hPG with a catecholic functional degree of 10% (hPG-Cat10) and hPG with both catecholic and vinylic functional degree of 5% (hPG-Cat5-V5) were identified as the best catecholic hPGs to prepare bioinert and stable monolayer coatings on TiO2. PMID:25189471

  20. Raft-based interactions of gangliosides with a GPI-anchored receptor.

    Komura, Naoko; Suzuki, Kenichi G N; Ando, Hiromune; Konishi, Miku; Koikeda, Machi; Imamura, Akihiro; Chadda, Rahul; Fujiwara, Takahiro K; Tsuboi, Hisae; Sheng, Ren; Cho, Wonhwa; Furukawa, Koichi; Furukawa, Keiko; Yamauchi, Yoshio; Ishida, Hideharu; Kusumi, Akihiro; Kiso, Makoto

    2016-06-01

    Gangliosides, glycosphingolipids containing one or more sialic acid(s) in the glyco-chain, are involved in various important physiological and pathological processes in the plasma membrane. However, their exact functions are poorly understood, primarily because of the scarcity of suitable fluorescent ganglioside analogs. Here, we developed methods for systematically synthesizing analogs that behave like their native counterparts in regard to partitioning into raft-related membrane domains or preparations. Single-fluorescent-molecule imaging in the live-cell plasma membrane revealed the clear but transient colocalization and codiffusion of fluorescent ganglioside analogs with a fluorescently labeled glycosylphosphatidylinisotol (GPI)-anchored protein, human CD59, with lifetimes of 12 ms for CD59 monomers, 40 ms for CD59's transient homodimer rafts in quiescent cells, and 48 ms for engaged-CD59-cluster rafts, in cholesterol- and GPI-anchoring-dependent manners. The ganglioside molecules were always mobile in quiescent cells. These results show that gangliosides continually and dynamically exchange between raft domains and the bulk domain, indicating that raft domains are dynamic entities. PMID:27043189

  1. Biomimetic anchors applied to the host-guest antifouling functionalization of titanium substrates.

    Cai, Xiao Yan; Li, Ning Ning; Chen, Jiu Cun; Kang, En-Tang; Xu, Li Qun

    2016-08-01

    A biomimetic strategy was developed for the construction of antifouling titanium oxide (Ti(oxide)) surfaces based on host-guest interactions. Two catecholic derivatives, dopamine 4-(phenylazo)benzamide (AZODopa) and dopamine 1-adamantanecarboxamide (AdaDopa) were synthesized and immobilized onto the Ti(oxide) surfaces. The guest molecules-anchored Ti(oxide) surfaces were further functionalized with zwitterionic heptakis[6-deoxy-6-(N-3-sulfopropyl-N,N-dimethylammonium ethyl sulfanyl)]-β-cyclodextrin (SBCD) and hydrophilic β-CD polymer (CDP). The surface elemental compositions and hydrophobic/hydrophilic properties of the Ti(oxide) surfaces before and after modification were characterized by X-ray photoelectron spectroscopy (XPS) and static water contact angle measurements, respectively. The antifouling properties of the modified Ti(oxide) surfaces were evaluated by the protein adsorption and bacterial adhesion assays. The zwitterionic SBCD- and hydrophilic CDP-functionalized Ti(oxide) surfaces can reduce the adsorption of bovine plasma fibrinogen and adhesion of Escherichia coli, as compared to the pristine and guest molecules-anchored Ti(oxide) surfaces. PMID:27135943

  2. Drosophila acetylcholinesterase: demonstration of a glycoinositol phospholipid anchor and an endogenous proteolytic cleavage

    The presence of a glycoinositol phospholipid anchor Drosophila acetylcholinesterase (AChE) was shown by several criteria. Chemical analysis of highly purified Drosophila AChE demonstrated approximately one residue of inositol per enzyme subunit. Selective cleavage by Staphylococcus aureus phosphatidylinositol-specific phospholipase C (PI-PLC) was tested with Drosophila AChE radiolabeled by the photoactivatable affinity probe 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine ([125I]TID), a reagent that specifically labels the lipid moiety of glycoinositol phospholipid-anchored proteins. Digestion with PI-PLC released 75% of this radiolabel from the protein. Gel electrophoresis of Drosophila AChE in sodium dodecyl sulfate indicated prominent 55- and 16-kDa bands and a faint 70-kDa band. The [125]I]TID label was localized on the 55-kDa fragment, suggesting that this fragment is the C-terminal portion of the protein. In support of this conclusion, a sensitive microsequencing procedure that involved manual Edman degradation combined with radiomethylation was used to determine residues 2-5 of the 16-kDa fragment. Comparison with the Drosophila AChE cDNA sequence confirmed that the 16-kDa fragment includes the N-terminus of AChE. Furthermore, the position of the N-terminal amino acid of the mature Drosophila AChE is closely homologous to that of Torpedo AChE. The presence of radiomethylatable ethanolamine in both 16- and 55-kDa fragments was also confirmed. Thus, Drosophila AChE may include a second posttranslational modification involving ethanolamine

  3. Iron anchors of Northern Odisha, east coast India: Maritime contacts with European countries

    Tripati, S.; Patnaik, S.K.; Acharya, S.B.

    at Siraf. Second Interim Report. Iran 8: 1–17. 12    List of Figures Figure 1. Iron anchor find sites, ports and trade centres of Odisha. Figure 2. Iron anchor found at Balaramgadi, Northern Odisha (Photo: Sila Tripati) Figure 2a. Iron anchor found... at Balaramgadi, Northern Odisha (S.B. Chitari & Rudra Behera). Figure 3. Iron anchor found at Chandabali on the mouth of the River Baitarani (OMISEAS, Bhubaneswar). Figure 4. Iron anchor found at Chandabali on the mouth of the River Baitarani (Photo: Sila...

  4. Graphene nanoribbons anchored to SiC substrates

    Le, Nam B.; Woods, Lilia M.

    2016-09-01

    Graphene nanoribbons are quasi-one-dimensional planar graphene allotropes with diverse properties dependent on their width and types of edges. Graphene nanoribbons anchored to substrates is a hybrid system, which offers novel opportunities for property modifications as well as experimental control. Here we present electronic structure calculations of zigzag graphene nanoribbons chemically attached via the edges to the Si or C terminated surfaces of a SiC substrate. The results show that the edge characteristics are rather robust and the properties are essentially determined by the individual nanoribbon. While the localized spin polarization of the graphene nanoribbon edge atoms is not significantly affected by the substrate, secondary energy gaps in the highest conduction and lowest valence region may emerge in the anchored structures. The van der Waals interaction together with the electrostatic interactions due to the polarity of the surface bonds are found to be important for the structure parameters and energy stability.

  5. Mobile Anchor Assisted Node Localization for Wireless Sensor Networks

    Chen, Hongyang; Huang, Pei; Poor, H Vincent; Sezaki, Kaoru

    2009-01-01

    In this paper, a cooperative localization algorithm is proposed that considers the existence of obstacles in mobilityassisted wireless sensor networks (WSNs). In this scheme, a mobile anchor (MA) node cooperates with static sensor nodes and moves actively to refine location performance. The localization accuracy of the proposed algorithm can be improved further by changing the transmission range of mobile anchor node. The algorithm takes advantage of cooperation betweenMAs and static sensors while, at the same time, taking into account the relay node availability to make the best use of beacon signals. For achieving high localization accuracy and coverage, a novel convex position estimation algorithm is proposed, which can effectively solve the localization problem when infeasible points occur because of the effects of radio irregularity and obstacles. This method is the only range-free based convex method to solve the localization problem when the feasible set of localization inequalities is empty. Simulatio...

  6. Simulation study of liquid crystal anchoring at a polymer surface

    Hamaneh, Mehdi

    2005-03-01

    The process of rubbing a polymer substrate to induce planar anchoring has two effects. It aligns the polymer chains and also creates grooves in the surface. We have investigated which one of these effects is more important by conducting a series of simulations of molecules of 5CB in contact with a poly(vinyl alcohol) surface. The polymer surface was constructed from a set of parallel straight chains. It was then distorted to mimic the effect of grooves in a direction perpendicular to the chain direction, thus causing two opposing anchoring effects. It was found that the 5CB molecules ordered preferentially along the chain direction when the depth of the grooves was less than 20 percent of the distance between grooves. For grooves whose walls were more steeply pitched, the nematic ordering aligned with the grooves.

  7. Organization, 'Anchoring' of Knowledge, and Innovative Activity in Construction

    Drejer, Ina; Vinding, Anker Lund

    2006-01-01

    The construction industry is characterised by the widespread use of project organisation. It has been suggested that the relatively low level of innovative activity in the industry can be explained by the temporary nature of firm boundary-crossing projects. Survey data from the Danish construction...... industry is used to investigate the importance of learning and 'anchoring' of project-specific knowledge at the firm level for participation in innovative activities. The data cover both the overall Danish construction industry and a specific region, North Jutland, which has a relatively high...... specialisation of construction workers. Latent class and regression analyses reveal that firms that make extensive use of partnering, together with internal product and process evaluation and knowledge diffusion (labelled 'knowledge-anchoring mechanisms'), are more likely to participate in innovative activities...

  8. Anchoring FRP Composite Armor in Flexible Offshore Riser Systems

    Costache, Andrei

    Unbonded flexible pipes find extensive use in the offshore oil industry. Although more expensive than rigid pipe, the total cost of flexible pipe installations are often less. This is because flexible pipes are easier to store and deploy, coupled with superior fatigue performance. Among other...... composite materials. One of the problems related to the substitution of tensile steel members is that anchoring in the metallic end fittings of the pipe is very challenging.The purpose of this thesis is to ensure the transfer of tensile loads between a unidirectional fiber reinforced polymer and a metallic...... the grip, and to measure the contact stresses distribution and evolution during pullout.The results of the experimental and numerical analysis show that it is possible to reliably anchor composite materials using a metallic grip. The models developed during the project show how to improve the...

  9. Graphene nanoribbons anchored to SiC substrates.

    Le, Nam B; Woods, Lilia M

    2016-09-14

    Graphene nanoribbons are quasi-one-dimensional planar graphene allotropes with diverse properties dependent on their width and types of edges. Graphene nanoribbons anchored to substrates is a hybrid system, which offers novel opportunities for property modifications as well as experimental control. Here we present electronic structure calculations of zigzag graphene nanoribbons chemically attached via the edges to the Si or C terminated surfaces of a SiC substrate. The results show that the edge characteristics are rather robust and the properties are essentially determined by the individual nanoribbon. While the localized spin polarization of the graphene nanoribbon edge atoms is not significantly affected by the substrate, secondary energy gaps in the highest conduction and lowest valence region may emerge in the anchored structures. The van der Waals interaction together with the electrostatic interactions due to the polarity of the surface bonds are found to be important for the structure parameters and energy stability. PMID:27392014

  10. Cell wall antibiotics provoke accumulation of anchored mCherry in the cross wall of Staphylococcus aureus.

    Wenqi Yu

    Full Text Available A fluorescence microscopy method to directly follow the localization of defined proteins in Staphylococcus was hampered by the unstable fluorescence of fluorescent proteins. Here, we constructed plasmid (pCX encoded red fluorescence (RF mCherry (mCh hybrids, namely mCh-cyto (no signal peptide and no sorting sequence, mCh-sec (with signal peptide, and mCh-cw (with signal peptide and cell wall sorting sequence. The S. aureus clones targeted mCh-fusion proteins into the cytosol, the supernatant and the cell envelope respectively; in all cases mCherry exhibited bright fluorescence. In staphylococci two types of signal peptides (SP can be distinguished: the +YSIRK motif SP(lip and the -YSIRK motif SP(sasF. mCh-hybrids supplied with the +YSIRK motif SP(lip were always expressed higher than those with -YSIRK motif SP(sasF. To study the location of the anchoring process and also the influence of SP type, mCh-cw was supplied on the one hand with +YSIRK motif (mCh-cw1 and the other hand with -YSIRK motif (mCh-cw2. MCh-cw1 preferentially localized at the cross wall, while mCh-cw2 preferentially localized at the peripheral wall. Interestingly, when treated with sub-lethal concentrations of penicillin or moenomycin, both mCh-cw1 and mCh-cw2 were concentrated at the cross wall. The shift from the peripheral wall to the cross wall required Sortase A (SrtA, as in the srtA mutant this effect was blunted. The effect is most likely due to antibiotic mediated increase of free anchoring sites (Lipid II at the cross wall, the substrate of SrtA, leading to a preferential incorporation of anchored proteins at the cross wall.