المؤلفون: Areti Gkourtsa, Teja Avula, Frans Hochstenbach, Janny van den Burg, Ben Distel

المساهمون: Other departments, Medical Biochemistry

المصدر: Microbiological research, 190, 27-36. Urban und Fischer Verlag Jena

مصطلحات موضوعية: Models, Molecular, 0301 basic medicine, Proline, Arginine, Protein Conformation, DNA Mutational Analysis, Molecular Sequence Data, 030106 microbiology, macromolecular substances, Microbiology, SH3 domain, Fungal Proteins, src Homology Domains, 03 medical and health sciences, Candida albicans, Amino Acid Sequence, Peptide ligand, chemistry.chemical_classification, biology, Tryptophan, biology.organism_classification, Amino acid, 030104 developmental biology, chemistry, Biochemistry, Hydrophobic motif, Protein Binding

الوصف: Src-homology 3 (SH3) domains are small protein-protein interaction modules. While most SH3 domains bind to proline-x-x-proline (PxxP) containing motifs in their binding partners, some SH3 domains recognize motifs other than proline-based sequences. Recently, we showed that the SH3 domain of Candida albicans Rvs167-3 binds peptides enriched in hydrophobic residues and containing a single proline residue (RΦxΦxΦP, where x is any amino acid and Φ is a hydrophobic residue). Here, we demonstrate that the proline in this motif is not required for Rvs167-3 SH3 recognition. Through mutagenesis studies we show that binding of the peptide ligand involves the conserved tryptophan in the canonical PxxP binding pocket as well as residues in the extended n-Src loop of Rvs167-3 SH3. Our studies establish a novel, proline-independent, binding sequence for Rvs167-3 SH3 (RΦxΦxΦ) that is comprised of a positively charged residue (arginine) and three hydrophobic residues.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d1d08074a11f27e8a772244b5aa77bfaTest

المؤلفون: Kazuya Machida, Wojciech Jankowski, Canan Kasikara, Tong Liu, Raymond B. Birge, Jing Li, Hong Li, Khanh-Quynh N. Nguyen, Charalampos G. Kalodimos, Charles Reichman, Peter Hornbeck, Ganapathy Sriram, Tamjeed Saleh

المصدر: Oncogene

مصطلحات موضوعية: Cancer Research, animal structures, Blotting, Western, Molecular Sequence Data, non-canonical signaling, Biology, SH2 domain, Article, SH3 domain, src Homology Domains, Tyrosine phosphorylation, Mice, Adapter molecule crk, chemistry.chemical_compound, Tandem Mass Spectrometry, Cell Line, Tumor, Genetics, Animals, Humans, Amino Acid Sequence, Phosphorylation, Oncogene Proteins v-abl, Molecular Biology, Sequence Homology, Amino Acid, Proto-Oncogene Proteins c-crk, Cell biology, HEK293 Cells, chemistry, atypical Crk SH3 domain, NIH 3T3 Cells, Tyrosine, Signal transduction, Chromatography, Liquid, Protein Binding, Signal Transduction, Proto-oncogene tyrosine-protein kinase Src

الوصف: Crk, the prototypical member of a class of Src homology-2 (SH2) and Src homology-3 (SH3) domain containing proteins that controls the coordinated assembly of signaling complexes, is regulated by phosphorylation of Y221 in the linker region, which forms an intramolecular SH2-pY221 auto-clamp to interrupt SH2-N-terminal SH3 domain (SH3N) signaling. Here, we show using LC-MS/MS and by generating phospho-specific antibodies that, iteratively with Y221, the Crk C-terminal SH3 domain (SH3C) is routinely phosphorylated on Y239 and/or Y251 by several extracellular stimuli known to engage Crk. Although phosphorylation at Y221 auto-inhibits the Crk SH2, phosphorylation of the SH3C generates an unconventional phosphoSH3C-SH3N unit in which the SH3N is fully functional to bind polyproline type II ligands and the phosphoSH3C binds de novo to other SH2 domains. Using high-throughput SH2 domain profiling, artificial neural network and position-specific scoring matrix-based bioinformatics approaches, and unbiased mass spectometry, we found that the phosphoSH3C binds several SH2 domain containing proteins, including specific non-receptor tyrosine kinases-Abl via pY251 and C-terminal Src kinase via pY239. Functionally, we show that the phosphoSH3C modulates the Abl-mediated phenotypes of cell spreading and motility. Together, these studies describe a versatile mechanism wherein phosphorylation of Crk at Y221 is not an off switch but redirects signaling from the SH2-SH3N axis to a phosphoSH3C-SH3N axis, with the SH3N as a common denominator.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9f8d13173df5032a3f55fdb57171e60dTest
https://doi.org/10.1038/onc.2014.361Test

المؤلفون: Françoise Guerlesquin, Matthieu Nouailler, Ali Badache, Corinne Sebban-Kreuzer, Michaël Feracci, Olivier Bornet, Xavier Morelli, Deborah Byrne, Hubert Halimi

المساهمون: Interactions et Modulateurs de Réponses (IMR), Centre National de la Recherche Scientifique (CNRS), Laboratoire d'ingénierie des systèmes macromoléculaires (LISM), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Microbiologie de la Méditerranée (IMM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU)

المصدر: FEBS Letters
FEBS Letters, 2014, 588 (12), pp.2031--6. ⟨10.1016/j.febslet.2014.04.029⟩
FEBS Letters, Wiley, 2014, 588 (12), pp.2031--6. ⟨10.1016/j.febslet.2014.04.029⟩

مصطلحات موضوعية: Models, Molecular, Magnetic Resonance Spectroscopy, Receptor, ErbB-2, Amino Acid Motifs, Molecular Sequence Data, Biophysics, IDP, Breast Neoplasms, Biology, Mitogen-activated protein kinase kinase, SH2 domain, Biochemistry, Receptor tyrosine kinase, SH3 domain, src Homology Domains, ErbB2, Structural Biology, Cell Line, Tumor, Genetics, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Proline rich motifs, Amino Acid Sequence, skin and connective tissue diseases, Molecular Biology, Binding Sites, Cell Biology, src-Family Kinases, Src-kinase, ROR1, biology.protein, Cyclin-dependent kinase 9, GRB2, Peptides, SH3 domain and NMR spectroscopy, Proto-oncogene tyrosine-protein kinase Src

الوصف: International audience; Overexpression of the ErbB2 receptor tyrosine kinase is associated with most aggressive tumors in breast cancer patients and is thus one of the main investigated therapeutic targets. Human ErbB2 C-terminal domain is an unstructured anchor that recruits specific adaptors for signaling cascades resulting in cell growth, differentiation and migration. Herein, we report the presence of a SH3 binding motif in the proline rich unfolded ErbB2 C-terminal region. NMR analysis of this motif supports a PPII helix conformation and the binding to Fyn-SH3 domain. The interaction of a kinase of the Src family with ErbB2 C-terminal domain could contribute to synergistic intracellular signaling and enhanced oncogenesis.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::62a281e1bca3b768b6b9e26de5187233Test
https://doi.org/10.1016/j.febslet.2014.04.029Test

المؤلفون: Michael Sporny, Naama Lahav-Mankovski, Yehezkel Sasson, Julia Guez-Haddad, Lada Gevorkyan-Airapetov, Jens Radzimanowski, Yarden Opatowsky, David Margulies

المصدر: Structure. 23:1989-2000

مصطلحات موضوعية: Binding Sites, GTPase-activating protein, Effector, GTPase-Activating Proteins, Molecular Sequence Data, Cell migration, Plasma protein binding, Biology, Ligands, SH3 domain, Substrate Specificity, Cell biology, Molecular Docking Simulation, src Homology Domains, Biochemistry, Structural Biology, Cytoplasm, Humans, Proline-Rich Protein Domains, Amino Acid Sequence, Binding site, Cytoskeleton, Molecular Biology, Protein Binding

الوصف: SummarysrGAP proteins regulate cell migration and morphogenesis by shaping the structure and dynamics of the cytoskeleton and membranes. First discovered as intracellular effectors for the Robo1 axon-guidance receptor, srGAPs were later identified as interacting with several other nuclear and cytoplasmic proteins. In all these cases, the srGAP SH3 domain mediates protein-protein interactions by recognizing a short proline-rich segment on the cognate-binding partner. However, as interactions between the isolated SH3 domain and a selected set of ligands show weak affinity and low specificity, it is not clear how srGAPs are precisely recruited to their signaling sites. Here, we report a two-component molecular mechanism that regulates ligand binding to srGAP2 by on the one hand dramatically tightening their association and on the other, moderately autoinhibiting and restricting binding. Our results allow the design of point mutations for better probing of srGAP2 activities, and may facilitate the identification of new srGAP2 ligands.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::68206b72fe8562477abc9585fc503204Test
https://doi.org/10.1016/j.str.2015.08.009Test

المؤلفون: Joe Xie, Qiqi Ye, Xiaoyu Cui, Sandrine V. Pierre, Qiying Yi, Namrata Madan, Zijian Xie

المصدر: American Journal of Physiology-Cell Physiology. 309:C373-C382

مصطلحات موضوعية: Cell signaling, MAP Kinase Signaling System, Swine, Physiology, Proto-Oncogene Proteins c-akt, Caveolin 1, Molecular Sequence Data, Down-Regulation, Ion Pumps, Biology, Kidney, SH3 domain, Cell Line, Animals, Amino Acid Sequence, Na+/K+-ATPase, Ouabain, Protein kinase B, Tyrosine-protein kinase CSK, Epithelial Cells, Cell Biology, Molecular biology, Rats, Cell biology, src-Family Kinases, Call for Papers, Sodium-Potassium-Exchanging ATPase, Signal transduction, Signal Transduction, Proto-oncogene tyrosine-protein kinase Src

الوصف: Na-K-ATPase is a fundamental component of ion transport. Four α isoforms of the Na-K-ATPase catalytic α subunit are expressed in human cells. The ubiquitous Na-K-ATPase α1 was recently discovered to also mediate signal transduction through Src kinase. In contrast, α2 expression is limited to a few cell types including myocytes, where it is coupled to the Na+/Ca2+exchanger. To test whether rat Na-K-ATPase α2 is capable of cellular signaling like its α1 counterpart in a recipient mammalian system, we used an α1 knockdown pig renal epithelial cell (PY-17) to create an α2-expressing cell line with no detectable level of α1 expression. These cells exhibited normal ouabain-sensitive ATPase, but failed to effectively regulate Src. In contrast to α1-expressing cells, ouabain did not stimulate Src kinase or downstream effectors such as ERK and Akt in α2 cells, although their signaling apparatus was intact as evidenced by EGF-mediated signal transduction. Additionally, α2 cells were unable to rescue caveolin-1. Unlike the NaKtide sequence derived from Na-K-ATPase α1, which downregulates basal Src activity, the corresponding α2 NaKtide was unable to inhibit Src in vitro. Finally, coimmunoprecipitation of cellular Src was diminished in α2 cells. These findings indicate that Na-K-ATPase α2 does not regulate Src and, therefore, may not serve the same role in signal transduction as α1. This further implies that the signaling mechanism of Na-K-ATPase is isoform specific, thereby supporting a model where α1 and α2 isoforms play distinct roles in mediating contraction and signaling in myocytes.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::89d13d6493dca90dbf7dec378cdb01a1Test
https://doi.org/10.1152/ajpcell.00103.2015Test

المؤلفون: Barbara Loesch, Joerg Kumbrink, Barbara Laumbacher, Kathrin H. Kirsch, Shefali Soni

المصدر: Journal of Biological Chemistry. 290:12247-12255

مصطلحات موضوعية: Cell signaling, animal structures, Molecular Sequence Data, macromolecular substances, Biology, environment and public health, Biochemistry, Gene Expression Regulation, Enzymologic, SH3 domain, src Homology Domains, Focal adhesion, Mice, Cell Movement, Sequence Homology, Nucleic Acid, Cell Adhesion, Animals, Humans, Protein Isoforms, Neoplasm Invasiveness, RNA, Messenger, Cell adhesion, Molecular Biology, Base Sequence, Alternative splicing, Cell migration, Exons, Cell Biology, Fibroblasts, Molecular biology, Protein Structure, Tertiary, Alternative Splicing, Crk-Associated Substrate Protein, Focal Adhesion Protein-Tyrosine Kinases, BCAR1, Signal Transduction

الوصف: Elevated levels of p130Cas (Crk-associated substrate)/BCAR1 (breast cancer antiestrogen resistance 1 gene) are associated with aggressiveness of breast tumors. Following phosphorylation of its substrate domain, p130Cas promotes the integration of protein complexes involved in multiple signaling pathways and mediates cell proliferation, adhesion, and migration. In addition to the known BCAR1-1A (wild-type) and 1C variants, we identified four novel BCAR1 mRNA variants, generated by alternative first exon usage (1B, 1B1, 1D, and 1E). Exons 1A and 1C encode for four amino acids (aa), whereas 1D and 1E encode for 22 aa and 1B1 encodes for 50 aa. Exon 1B is non-coding, resulting in a truncated p130Cas protein (Cas1B). BCAR1-1A, 1B1, and variant 1C mRNAs were ubiquitously expressed in cell lines and a survey of human tissues, whereas 1B, 1D, and 1E expression was more restricted. Reconstitution of all isoforms except for 1B in p130Cas-deficient murine fibroblasts induced lamellipodia formation and membrane ruffling, which was unrelated to the substrate domain phosphorylation status. The longer isoforms exhibited increased binding to focal adhesion kinase (FAK), a molecule important for migration and adhesion. The shorter 1B isoform exhibited diminished FAK binding activity and significantly reduced migration and invasion. In contrast, the longest variant 1B1 established the most efficient FAK binding and greatly enhanced migration. Our results indicate that the p130Cas exon 1 variants display altered functional properties. The truncated variant 1B and the longer isoform 1B1 may contribute to the diverse effects of p130Cas on cell biology and therefore will be the target of future studies. Background: The tumor promoter p130Cas utilizes its SH3 domain to control cell adhesion and migration. Results: We identified N-terminal extended p130Cas variants with enhanced SH3 domain binding activity which modulates cell migration and invasion. Conclusion: Naturally occurring p130Cas variants exhibit differential biological activities. Significance: Knowledge on these variants may explain how p130Cas controls cellular programs and drives carcinogenesis.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e0c4680db512a0a93f78557b67fa62e3Test
https://doi.org/10.1074/jbc.m115.649947Test

المؤلفون: Caitlin A. Ruddick, Wei Jia, Mandi M. Murph, Sterling K. Tran

المصدر: Cancer Letters. 356:589-596

مصطلحات موضوعية: Cancer Research, Blotting, Western, Molecular Sequence Data, Fluorescent Antibody Technique, Apoptosis, Biology, SH3 domain, src Homology Domains, chemistry.chemical_compound, Valine, Lysophosphatidic acid, Tumor Cells, Cultured, Humans, Amino Acid Sequence, Receptors, Lysophosphatidic Acid, Receptor, Melanoma, Cell Proliferation, Alanine, chemistry.chemical_classification, Sequence Homology, Amino Acid, Molecular biology, Amino acid, Oncology, chemistry, Mutation, Mutagenesis, Site-Directed, Calcium, Proto-oncogene tyrosine-protein kinase Src, Binding domain

الوصف: The LPA3 receptor is a G protein-coupled receptor that binds extracellular lysophosphatidic acid and mediates intracellular signaling cascades. Although we previously reported that receptor inhibition using siRNA or chemical inhibition obliterates the viability of melanoma cells, the mechanism was unclear. Herein we hypothesized that amino acids comprising the Src homology 3 (SH3) ligand binding motif, R/K-X-X-V/P-X-X-P or (216)-KTNVLSP-(222), within the third intracellular loop of LPA3 were critical in mediating this outcome. Therefore, we performed site-directed mutagenesis of the lysine, valine and proline, replacing these amino acids with alanines, and evaluated the changes in viability, proliferation, ERK1/2 signaling and calcium in response to lysophosphatidic acid. Our results show that enforced LPA3 expression in SK-MEL-2 cells enhanced their resiliency by allowing these cells to oppose any loss of viability during growth in serum-free medium for up to 96 h, in contrast to parental SK-MEL-2 cells, which show a significant decline in viability. Similarly, site-directed alanine substitutions of valine and proline, V219A/P222A or 2aa-SK-MEL-2 cells, did not significantly alter viability, but adding a further alanine to replace the lysine, K216A/V219A/P222A or 3aa-SK-MEL-2 cells, obliterated this function. In addition, an inhibitor of the LPA3 receptor had no impact on the parental SK-MEL-2, 2aa-SK-MEL-2 or 3aa-SK-MEL-2 cells, but significantly reduced viability among wt-LPA3-SK-MEL-2 cells. Taken together, the data suggest that the SH3 ligand binding domain of LPA3 is required to mediate viability in melanoma cells.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::58c137b8cdee1e1545ab54484998fb12Test
https://doi.org/10.1016/j.canlet.2014.10.001Test

المؤلفون: Oliver Seitz, J. Schmalisch

المصدر: Chemical Communications. 51:7554-7557

مصطلحات موضوعية: chemistry.chemical_classification, Stereochemistry, Molecular Sequence Data, Metals and Alloys, General Chemistry, Native chemical ligation, Catalysis, SH3 domain, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, src Homology Domains, chemistry, Intramolecular force, Materials Chemistry, Ceramics and Composites, Thiol, Reactivity (chemistry), Amino Acid Sequence, Cysteine, Sulfhydryl Compounds, Peptides, Peptide sequence

الوصف: The old dog and a new trick; mercaptopropionylcysteine (MPA–Cys) peptide thioesters show a surprisingly high reactivity in native chemical ligation (NCL) and allow thiol-additive free reactions. This facilitates sequential NCL reactions and ligation–desulfurization reactions in one-pot formats. The synthetic utility is demonstrated by the synthesis of a SH3 domain.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b89871077a99d85ec9aa850477959a9bTest
https://doi.org/10.1039/c5cc01447fTest

المؤلفون: Tingting Wu, Tobias Baumgart

المصدر: Biochemistry

مصطلحات موضوعية: Molecular Sequence Data, Biology, Phosphatidylinositols, Biochemistry, Dynamin II, Article, SH3 domain, Cell Line, src Homology Domains, Cell membrane, medicine, Humans, BAR domain, Amino Acid Sequence, Protein Interaction Maps, Adaptor Proteins, Signal Transducing, Dynamin, Tumor Suppressor Proteins, Cell Membrane, Nuclear Proteins, Cell biology, Membrane, medicine.anatomical_structure, Membrane curvature, Liposomes, Proto-oncogene tyrosine-protein kinase Src

الوصف: In striated muscles, invaginations from the plasma membrane, termed transverse tubules (T-tubule), function in the excitation–contraction coupling machinery. BIN1 (isoform8) plays a critical role in the biogenesis of T-tubules. BIN1 contains an N-terminal BAR domain to sense and induce membrane curvature, an isoform8-specific polybasic motif (exon10) as the phosphoinositide binding module and a C-terminal Src homology 3 (SH3) domain for the recruitment of downstream proteins such as dynamin 2. Previous studies of N-BAR domains focused on elucidating mechanisms of membrane curvature sensing and generation (MC-S&G). Less is known about how MC-S&G is regulated. We found that the SH3 domain binds to the exon10 motif more strongly compared to the proline-rich domain (PRD) of dynamin 2. Furthermore, we found that the MC-S&G ability of full-length BIN1 is inhibited on membranes lacking PI(4,5)P2. Addition of PI(4,5)P2 in the membrane activates BIN1 to sense and induce membrane curvature. Co-presence of the SH3 domain and exon10 motif leads to the strongest phosphoinositide-mediated control of BIN1 function. Addition of SH3 domain ligand (such as PRD peptides), as well as addition of the water-soluble PI(4,5)P2 analogue, can both enhance the MC-S&G ability of BIN1 on membranes without PI(4,5)P2, indicating that the key to activate BIN1 is to disrupt the exon10–SH3 interaction. The nonsense mutation K436X, found in centronuclear myopathy (CNM) patients, abolishes SH3 domain binding with either exon10 or the PRD motif, resulting in increased membrane deformation capacity. Our results suggest an autoinhibition model for BIN1 that involves a synergistic regulation by membrane composition and protein–protein interactions.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9386aa4c1353e131451e87ea70186327Test
https://doi.org/10.1021/bi501082rTest

المؤلفون: Debashis Mukhopadhyay, Sreetama Basu, Shounak Baksi

المصدر: Neuroscience Research. 87:77-83

مصطلحات موضوعية: congenital, hereditary, and neonatal diseases and abnormalities, Huntingtin, MAP Kinase Signaling System, Molecular Sequence Data, Mutant, GAB1, Nerve Tissue Proteins, SH3 domain, src Homology Domains, Mice, Growth factor receptor binding, Cell Line, Tumor, mental disorders, Animals, Humans, Adaptor Proteins, Signal Transducing, GRB2 Adaptor Protein, Huntingtin Protein, Base Sequence, biology, Kinase, General Neuroscience, Signal transducing adaptor protein, General Medicine, Molecular biology, nervous system diseases, nervous system, biology.protein, GRB2, biological phenomena, cell phenomena, and immunity, Protein Binding

الوصف: Huntington's disease (HD) is caused due to expansion of CAG repeats in the gene huntingtin (Htt). Adaptor protein Grb2, involved in Ras-MAP kinase pathway, is a known interactor of Htt. Mutant Htt-Grb2 interaction reduces Ras-MAPK signaling in HD models. In normal cells Grb2 forms Grb2-Sos1-Gab1 complex through its N-SH3 and C-SH3 domains respectively, essential for sustained activation of Ras. We found that C-SH3 of Grb2 mediates the interaction with mutant Htt and this interaction being stronger could replace Gab1, with mutant Htt becoming the preferred partner. This would have immense effect on downstream signaling events.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fcc9950da445d7f84a43bffa72716e05Test
https://doi.org/10.1016/j.neures.2014.06.009Test