排序方式: 共有342条查询结果,搜索用时 125 毫秒
71.
Emeline Assémat Emmanuelle Crost Marion Ponserre Jan Wijnholds Andre Le Bivic Dominique Massey-Harroche 《Experimental cell research》2013
MUPP-1 (multi-PDZ domain protein-1) and PATJ (PALS-1-associated tight junction protein) proteins are closely related scaffold proteins and bind to many common interactors including PALS-1 (protein associated with Lin seven) a member of the Crumbs complex. Our goal is to understand how MUPP-1 and PATJ and their interaction with PALS-1 are regulated in the same cells. We have shown that in MCF10A cells there are at least two different and co-existing complexes, PALS-1/MUPP-1 and PALS-1/PATJ. Surprisingly, MUPP-1 levels inversely correlated with PATJ protein levels by acting on the stabilization of the PATJ/PALS-1 complex. Upon MUPP-1 depletion, the increased amounts of PATJ are in part localized at the migrating front of MCF10A cells and are able to recruit more PAR3 (partition defective 3). All together these data indicate that a precise balance between MUPP-1 and PATJ is achieved in epithelial cells by regulating their association with PALS-1. 相似文献
72.
HtrA2/Omi is a mammalian mitochondrial serine protease, and was found to have dual roles in mammalian cells, not only acting
as an apoptosis-inducing protein but also maintaining mitochondrial homeostasis. PDZ domain is one of the most important protein-protein
interaction modules and is involved in a variety of important cellular functions, such as signal transduction, degradation
of proteins, and formation of cytoskeleton. Recently, it was reported that the PDZ domain of HtrA2/Omi might regulate proteolytic
activity through its interactions with ligand proteins. In this study, we rapidly characterized the binding properties of
HtrA2/Omi PDZ domain by validation screening of the PDZ ligand library with yeast two-hybrid approach. Then, we predicted
its novel ligand proteins in human proteome and reconfirmed them in the yeast two-hybrid system. Finally, we analyzed the
smallest networks bordered by the shortest path length between the protein pairs of novel interactions to evaluate the confidence
of the identified interactions. The results revealed some novel binding properties of HtrA2/Omi PDZ domain. Besides the reported
Class II PDZ motif, it also binds to Class I and Class III motifs, and exhibits restricted variability at P−3, which means that the P−3 residue is selected according to the composition of the last three residues. Seven novel ligand proteins of HtrA2/Omi PDZ
domain were discovered, providing significant clues for further clarifying the roles of HtrA2/Omi. Moreover, this study proves
the high efficiency and practicability of the newly developed validation screening of candidate ligand library method for
binding property characterization of peptide-binding domains. 相似文献
73.
74.
Interactions between proteins are an essential part of biology, and the desire to identify these interactions has led to the development of numerous technologies to systematically map protein–protein interactions at a large scale. As in most cellular processes, protein interactions are central to the control of cell polarity, and a full understanding of polarity will require comprehensive knowledge of the protein interactions involved. At its core, cell polarity is established through carefully regulated mutually inhibitory interactions between several groups of cortical proteins. While several interactions have been identified, the dynamics and molecular mechanisms that control these interactions are not well understood. Cell polarity also needs to be integrated with cellular processes including junction formation, cytoskeletal organization, organelle positioning, protein trafficking, and functional specialization of membrane domains. Moreover, polarized cells need to respond to external cues that coordinate polarity at the tissue level. Identifying the protein–protein interactions responsible for integrating polarity with all of these processes remains a major challenge, in part because the mechanisms of polarity control vary in different contexts and with developmental times. Because of their unbiased nature, systematic large-scale protein–protein interaction mapping approaches can be particularly helpful to identify such mechanisms. Here, we discuss methods commonly used to generate proteome-wide interactome maps, with an emphasis on advances in our understanding of cell polarity that have been achieved through application of such methods. 相似文献
75.
Sarah Garrido-Urbani Alain Vonlaufen Jimmy Stalin Maria De Grandis Patricia Ropraz Stéphane Jemelin Florence Bardin Holger Scheib Michel Aurrand-Lions Beat A. Imhof 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》2018,1865(4):638-649
Most cancer deaths result from metastasis, which is the dissemination of cells from a primary tumor to distant organs. Metastasis involves changes to molecules that are essential for tumor cell adhesion to the extracellular matrix and to endothelial cells. Junctional Adhesion Molecule C (JAM-C) localizes at intercellular junctions as homodimers or more affine heterodimers with JAM-B. We previously showed that the homodimerization site (E66) in JAM-C is also involved in JAM-B binding. Here we show that neoexpression of JAM-C in a JAM-C-negative carcinoma cell line induced loss of adhesive property and pro-metastatic capacities. We also identify two critical structural sites (E66 and K68) for JAM-C/JAM-B interaction by directed mutagenesis of JAM-C and studied their implication on tumor cell behavior. JAM-C mutants did not bind to JAM-B or localize correctly to junctions. Moreover, mutated JAM-C proteins increased adhesion and reduced proliferation and migration of lung carcinoma cell lines. Carcinoma cells expressing mutant JAM-C grew slower than with JAM-C WT and were not able to establish metastatic lung nodules in mice. Overall these data demonstrate that the dimerization sites E66-K68 of JAM-C affected cell adhesion, polarization and migration and are essential for tumor cell metastasis. 相似文献
76.
Christoph Klenk Thorsten Vetter Alexander Zürn Jean-Pierre Vilardaga Peter A. Friedman Bin Wang Martin J. Lohse 《The Journal of biological chemistry》2010,285(39):30355-30362
β-Arrestins are crucial regulators of G-protein coupled receptor (GPCR) signaling, desensitization, and internalization. Despite the long-standing paradigm that agonist-promoted receptor phosphorylation is required for β-arrestin2 recruitment, emerging evidence suggests that phosphorylation-independent mechanisms play a role in β-arrestin2 recruitment by GPCRs. Several PDZ proteins are known to interact with GPCRs and serve as cytosolic adaptors to modulate receptor signaling and trafficking. Na+/H+ exchange regulatory factors (NHERFs) exert a major role in GPCR signaling. By combining imaging and biochemical and biophysical methods we investigated the interplay among NHERF1, β-arrestin2, and the parathyroid hormone receptor type 1 (PTHR). We show that NHERF1 and β-arrestin2 can independently bind to the PTHR and form a ternary complex in cultured human embryonic kidney cells and Chinese hamster ovary cells. Although NHERF1 interacts constitutively with the PTHR, β-arrestin2 binding is promoted by receptor activation. NHERF1 interacts directly with β-arrestin2 without using the PTHR as an interface. Fluorescence resonance energy transfer studies revealed that the kinetics of PTHR and β-arrestin2 interactions were modulated by NHERF1. These findings suggest a model in which NHERF1 may serve as an adaptor, bringing β-arrestin2 into close proximity to the PTHR, thereby facilitating β-arrestin2 recruitment after receptor activation. 相似文献
77.
HtrA2/Omi is a mammalian mitochondrial serine protease, and was found to have dual roles in mammalian cells, not only acting as an apoptosis-inducing protein but also maintaining mitochondrial homeostasis. PDZ domain is one of the most important protein-protein interaction modules and is involved in a variety of important cellular functions, such as signal transduction, degradation of proteins,and formation of cytoskeleton. Recently, it was reported that the PDZ domain of HtrA2/Omi might regulate proteolytic activity through its interactions with ligand proteins. In this study, we rapidly characterized the binding properties of HtrA2/Omi PDZ domain by validation screening of the PDZ ligand library with yeast two-hybrid approach. Then, we predicted its novel ligand proteins in human proteome and reconfirmed them in the yeast two-hybrid system. Finally, we analyzed the smallest networks bordered by the shortest path length between the protein pairs of novel interactions to evaluate the confidence of the identified interactions. The results revealed some novel binding properties of HtrA2/Omi PDZ domain. Besides the reported Class Ⅱ PDZ motif, it also binds to Class Ⅰ and Class Ⅲ motifs, and exhibits restricted variability at P-3, which means that the P-3 residue is selected according to the composition of the last three residues. Seven novel ligand proteins of HtrA2/Omi PDZ domain were discovered, providing significant clues for further clarifying the roles of HtrA2/Omi.Moreover, this study proves the high efficiency and practicability of the newly developed validation screening of candidate ligand library method for binding property characterization of peptide-binding domains. 相似文献
78.
Gileadi O Knapp S Lee WH Marsden BD Müller S Niesen FH Kavanagh KL Ball LJ von Delft F Doyle DA Oppermann UC Sundström M 《Journal of structural and functional genomics》2007,8(2-3):107-119
As many of the structural genomics centers have ended their first phase of operation, it is a good point to evaluate the scientific
impact of this endeavour. The Structural Genomics Consortium (SGC), operating from three centers across the Atlantic, investigates
human proteins involved in disease processes and proteins from Plasmodium falciparum and related organisms. We present here some of the scientific output of the Oxford node of the SGC, where the target areas
include protein kinases, phosphatases, oxidoreductases and other metabolic enzymes, as well as signal transduction proteins.
The SGC has aimed to achieve extensive coverage of human gene families with a focus on protein–ligand interactions. The methods
employed for effective protein expression, crystallization and structure determination by X-ray crystallography are summarized.
In addition to the cumulative impact of accelerated delivery of protein structures, we demonstrate how family coverage, generic
screening methodology, and the availability of abundant purified protein samples, allow a level of discovery that is difficult
to achieve otherwise. The contribution of NMR to structure determination and protein characterization is discussed. To make
this information available to a wide scientific audience, a new tool for disseminating annotated structural information was
created that also represents an interactive platform allowing for a continuous update of the annotation by the scientific
community. 相似文献
79.
APC and PTEN are tumor suppressor proteins that bind through their C-termini to the PDZ domain containing-hDlg scaffolding protein. We have found that co-expression of PTEN and hDlg enhanced the negative regulation of the PI3K/Akt pathway by PTEN, indicating the physiologic importance of these interactions. APC and PTEN share other PDZ domain containing-interacting partners, including the MAGI scaffolding proteins and the MAST family of protein kinases. Mutational analysis revealed that the C-terminal PDZ-binding motifs from APC and PTEN were differentially recognized by distinct PDZ domains. APC bound to the three PDZ domains from hDlg, whereas PTEN mainly bound to PDZ-2/hDlg. This indicates the existence of overlapping, but distinct PDZ-domain recognition patterns by APC and PTEN. Furthermore, a ternary complex formed by APC, PTEN, and hDlg was detected, suggesting that hDlg may serve as a platform to bring in proximity APC and PTEN tumor suppressor activities. In line with this, tumor-related mutations targeting the PDZ-2/hDlg domain diminished its interaction with APC and PTEN. Our results expand the PDZ-domain counterparts for the tumor suppressor APC, show that APC and PTEN share PDZ-domain partners but have individual molecular determinants for specific recognition of PDZ domains, and suggest the participation of the tumor suppressors APC, PTEN, and hDlg in PDZ-domain interaction networks which may be relevant in oncogenesis. 相似文献
80.
Marie Gottschalk Anders Bach Jakob Lerche Hansen Povl Krogsgaard-Larsen Anders S. Kristensen Kristian Strømgaard 《Neurochemical research》2009,34(10):1729-1737
The PDZ domain mediated interaction between the NMDA receptor and its intracellular scaffolding protein, PSD-95, is a potential target for treatment of ischemic brain diseases. We have recently developed a number of peptide analogues with improved affinity for the PDZ domains of PSD-95 compared to the endogenous C-terminal peptide of the NMDA receptor, as evaluated by a cell-free protein–protein interaction assay. However, it is important to address both membrane permeability and effect in living cells. Therefore a bioluminescence resonance energy transfer (BRET) assay was established, where the C-terminal of the NMDA receptor and PDZ2 of PSD-95 were fused to green fluorescent protein (GFP) and Renilla luciferase (Rluc) and expressed in COS7 cells. A robust and specific BRET signal was obtained by expression of the appropriate partner proteins and subsequently, the assay was used to evaluate a Tat-conjugated peptide for its ability to disrupt the PSD-95/NMDA receptor interaction in living cells. 相似文献