Download or read book Characterization of beta arrestins Trafficking and Signaling Functions on G Protein coupled Receptors written by Etienne Khoury and published by . This book was released on 2015 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Characterization of arrestins Trafficking and Signaling Functions on G Protein coupled Receptors written by Etienne Khoury and published by . This book was released on 2015 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: "The heptahelical G protein-coupled receptors (GPCRs) are the largest, most versatile superfamily of cell surface receptors and constitute the most common target for many therapeutic drugs. They participate in various physiological processes via signaling transduction, mainly induced by heterotrimeric G proteins. Following activation, these 7-transmembrane receptors recruit [beta]-arrestin for endocytosis. Beta-arrestins will directlyassociate GPCRs to several components of the endocytic machinery, such as clathrin and adaptor protein 2 (AP-2), allowing the receptor to internalize. Over the years, [beta]-arrestins were shown to also act as signaling adaptors upon binding to agonist-occupied GPCRs; however, the mechanism regulating receptor/[beta]-arrestin complexes in endosomes was not yet addressed. Based on a previous study where we showed that [beta]-arrestin serves as a scaffolding protein for several signaling effectors (e.g. MAPK) in the endosomes, we hypothesized that endosomal MAPK activity would be necessary to maintain the GPCR/[beta]-arrestin-2 complex, thus regulating receptor trafficking. Our first study revealed a putative phosphorylating MAPK motif (Thr178) in [beta]-arrestin-2 suggesting that endosomal MAPK activity is involved in such process. Using biochemical assays and FRAP (Fluorescence Recovery After Photobleaching) approach to assess the life-time of bradykinin B2 receptor (B2R)/[beta]-arrestin-2 endosomal complexes, we showed that the MAPK putative site in [beta]-arrestin-2 is involved in regulating the association between the arrestin and its receptor. The role of [beta]-arrestin-2 'hinge' domain was tested for such effect, and using arrestin mutants demonstrated distinct behaviours between [beta]-arrestin-2 species on GPCRs trafficking and agonist mediated receptor signaling. This study highlights a strong correlation between MAPKsignaling and the regulation of endosomal GPCR/[beta]-arrestin-2 interactions. In addition to the 'hinge' domain, structural studies showed that the polar core, as well as the arrestins loops, were also crucial for [beta]-arrestin activation. Based on the crystal structure of [beta]-arrestin-1, a virtual screen was then conducted in order to identify aselective pharmacological inhibitor of [beta]-arrestin-2. Results showed a unique compound, namely UM0012685 which firstly, blocked V2-vasopressin receptor (V2R) endocytosis; Secondly, inhibited [beta]-arrestin recruitment; Thirdly, decreased the stability of the receptor and [beta]-arrestin-2 endosomal complexes and finally inhibited [beta]-arrestin-2-dependent MAPK activation upon agonist stimulation of the V2R. This compound was also used as a tool to determine [beta]-arrestins trafficking function on [beta]2-adrenergic receptor ([beta]2AR) recycling. These results uncover a better understanding of the underlying mechanism regulating [beta]-arrestin in the endosomes as well as the intracellular trafficking modulation of GPCRs. Our findings also reveal a useful tool to investigate the multifunctional aspect of [beta]-arrestins in the cell." --
Download or read book G Protein Coupled Receptors written by and published by Academic Press. This book was released on 2016-02-26 with total page 522 pages. Available in PDF, EPUB and Kindle. Book excerpt: G-Protein-Coupled Receptors: Signaling, Trafficking, and Regulation, a new volume in the Methods in Cell Biology series continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in G-Protein-Coupled Receptors, and includes sections on such topics signaling, trafficking and regulation. Covers the increasingly appreciated cell biology field of G-protein-coupled receptors Includes both established and new technologies Contributed by experts in the field Covers topics such as signaling, trafficking, and regulation
Download or read book Characterization of Beta arrestin PAR2 Interactions written by Puneet Kumar and published by . This book was released on 2008 with total page 282 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Trafficking of GPCRs written by Guangyu Wu and published by Academic Press. This book was released on 2015-06-05 with total page 351 pages. Available in PDF, EPUB and Kindle. Book excerpt: G protein-coupled receptors (GPCRs) constitute the largest superfamily of cell surface receptors that regulate a variety of cell functions. Over the past few decades great progress has been made in defining the roles of intracellular trafficking in controlling the functionality of the receptors as well as in the development of various human diseases. This volume of Progress in Molecular Biology and Translational Science reviews the recent understanding of GPCR trafficking regulators and molecular mechanisms. Written by future leaders in the pain field Covers a wide range of targets Contains provocative ideas about GPCR trafficking
Download or read book G Protein Coupled Receptors written by and published by Academic Press. This book was released on 2019-01-05 with total page 332 pages. Available in PDF, EPUB and Kindle. Book excerpt: G-Protein-Coupled Receptors, Part B, 2nd Edition, Volume 149, the latest release in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers Optical Approaches for Visualization of Arrestin Binding to Muscarinic Receptors, Luciferase Reporter Assay for Unlocking Ligand-mediated Signaling of GPCRs, Assays to Measure GPCR Dependent Cellular Migration, Characterization of the Frizzled GPCRs, Binding Assays for Bradykinin and Angiotensin Receptors, Detection of Misfolded Rhodopsin Aggregates in Cells, Measuring GPCR Ubiquitination and Trafficking, Culture of Primary Neurons and its Use in Studying GPCR Trafficking, and much more. Covers the increasingly appreciated cell biology field of G-protein-coupled receptors Includes both established and new technologies Contributed by experts in the field
Download or read book Nuclear G Protein Coupled Receptors written by Bruce G. Allen and published by Humana. This book was released on 2016-08-23 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nuclear G-Protein Coupled Receptors: Methods and Protocols is a compilation of a number of conceptual and methodological aspects important for the validation and characterization of intacrine signaling systems. To date, the best-characterized intracrine signaling system is that of angiotensin II (Ang II), covered in depth in various chapters. Methodology to study the subcellular localization and function of GPCRs and other signaling systems is provided, as well as numerous chapters focusing on methods designed to understand signaling mediated by nuclear and other internal GPCRs. Methods are also described to study the formation of second messengers such as cAMP and to study the trafficking of receptors from the cell surface. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Nuclear G-Protein Coupled Receptors: Methods and Protocols seeks to serve both professionals and novices with state-of-the-art approaches to characterize what is becoming a common theme in cellular signaling.
Download or read book Characterization of Protease activated Receptor 4 Trafficking and Heterodimerization in Modulating Receptor Signaling written by Thomas Horace Smith and published by . This book was released on 2016 with total page 140 pages. Available in PDF, EPUB and Kindle. Book excerpt: G protein-coupled receptors (GPCRs) are transmembrane proteins that allow cells to respond to extracellular stimuli. GPCR activation occurs when a ligand binds to the extracellular portion of the receptor. The ligand-bound receptor undergoes a conformational change that allows the intracellular domain of the receptor to engage and activate signaling effectors. The protease-activated receptors (PARs) are a family of GPCRs that are activated by proteases such as thrombin. Unlike traditional GPCRs, which can return to their inactive state after signaling, PARs are irreversibly activated. Therefore, the eventual fate of an activated PAR is degradation. The internalization and endocytic sorting of PARs play key roles in their signal regulation. Trafficking has been well-characterized for PAR1, the prototypical thrombin receptor. PAR4 was the last of the four thrombin receptor to be discovered, and has in large part been relatively under-studied. Recent studies have shown that PAR4 plays distinct roles that differ from those of PAR1, and as such may represent a potential drug target. Thus, understanding the mechanisms that regulate PAR4 signal regulation is important. In this dissertation, I characterized the role that trafficking and heterodimerization play in regulating PAR4 signaling. I found that adaptor-protein complex-2 (AP-2) is a key mediator of PAR4 agonist-induced internalization, and that AP-2 binds to intracellular loop 3 (ICL3) of PAR4. Disruption of PAR4 internalization resulted in enhanced and prolonged ERK1/2 activation, suggesting that endocytosis may mediate PAR4 signal attenuation. I also characterized the role of heterodimerization with the puringeric receptor P2Y12 in modulating PAR4 signaling. Previous studies had demonstrated that PAR4 and P2Y12 coordinate [beta]-arrestin-dependent Akt activation. My work strongly suggests that PAR4 and P2Y12 physically associate basally and co-internalize in response to activation of PAR4. I also demonstrate that [beta]-arrestin is recruited to the co-internalized PAR4-P2Y12 complex. In contrast to the effect on ERK1/2 activation, disruption of PAR4 internalization diminished Akt activation. Taken together, the studies summarized in this dissertation highlight the importance of PAR4 internalization in modulating activity of functionally and spatially distinct signaling pathways.
Download or read book Methods for the Discovery and Characterization of G Protein Coupled Receptors written by Craig W. Stevens and published by Humana Press. This book was released on 2011-07-15 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The goal of the characterization and discovery of G protein-coupled receptors, arguably the most important class of signaling molecules in humans and other vertebrates, has spawned numerous vital methodologies. In Methods for the Discovery and Characterization of G Protein-Coupled Receptors, experts in the field present the very latest on the methods and technology used to characterize and discover novel mechanisms of GPCRs which, in many cases, can be used directly to design experiments for the reader’s particular GPCR of interest and their specific avenue of investigation. Divided into four convenient sections, this detailed volume covers GPCRs in the genome, trafficking of GPCRs, GPCRs on the membrane, as well as the regulation of these key receptors. Chapters also feature an important section called “Future Directions” which gives the reader an insight into advances soon to be realized in each area. Written for the popular Neuromethods series, this book contains the kind of detailed description and implementation advice that is crucial for getting optimal results. Authoritative and cutting-edge, Methods for the Discovery and Characterization of G Protein-Coupled Receptors serves as an ideal guide for scientists determined to further our knowledge of crucially important set of receptors.
Download or read book G Protein Coupled Receptors written by and published by Academic Press. This book was released on 2013-01-24 with total page 489 pages. Available in PDF, EPUB and Kindle. Book excerpt: This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. This volume covers G protein coupled receptors and includes chapters on such topics as G protein-coupled receptor trafficking motifs, structure-based virtual screening, and automation-friendly high throughput assays for identification of pharmacoperone drugs. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers G protein coupled receptors Contains chapters on such topics as G protein-coupled receptor trafficking motifs, structure-based virtual screening, and automation-friendly high-throughput assays for identifying pharmacoperone drugs
Download or read book Beta Arrestins written by Mark G. H. Scott and published by Humana Press. This book was released on 2019-03-28 with total page 409 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume looks at various approaches to study the pleiotropic roles of b-arrestins (b-arrs) in the control of signal transduction, and the resulting cellular and in vivo consequences that arise. The chapters in this book cover diverse topics around b-arrs such as their established roles in GPCR regulation and trafficking; regulatory scaffolding functions of b-arrs in MAPK signaling, cAMP hydrolysis and cytoskeletal dynamics; proteomic analysis of the b-arr interactome; mathematical modelling of b-arr signaling networks; functional selectivity involving biased ligands; nucleocytoplasmic trafficking and primary cilia-associated functions of b-arrs; conformational plasticity of b-arrs; and the roles of b-arrs in allergic inflammation, Type 2 Diabetes, Parkinson’s disease, and cancer. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and authoritative, Beta-Arrestins: Methods and Protocols is a valuable resource for researchers interested in learning more about the function and regulation of b-arrestins.
Download or read book Characterization of a Small Molecule Inhibitor of Small G Proteins Regulating AT1R Trafficking and Signaling written by Jenna Giubilaro and published by . This book was released on 2020 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: "G protein–coupled receptors (GPCRs) are important therapeutic targets that activate various downstream signaling events, like mitogen-activated protein kinases (MAPK, for instance ERK1/2), via both G proteins and [beta]-arrestins. GPCR-mediated ERK1/2 signaling occurs both at the plasma membrane and in endosomes from receptor-[beta]-arrestin complexes. However, deciphering where ERK1/2 signals occur in cells, as well as the role(s) of [beta]-arrestin in such spatiotemporal signaling remains a challenge due to the dearth of tools. Here, using BRET-based sensors in a high-throughput screen for angiotensin II type 1 receptor (AT1R) trafficking, a small molecule, named Rasarfin, was identified that selectively inhibits small GTPases Ras and Arf6 but not heterotrimeric or other small G proteins. Rasarfin blocks agonist-promoted endocytosis of the AT1, Bradykinin B2 (B2R) and [beta]2-adrenergic ([beta]2AR) receptors and inhibits the activation of ERK1/2 by these receptors, including EGFR, as well as the activation of Akt. Molecular dynamics (MD) simulations reveal that Rasarfin binds Ras at the interface between Ras and its guanine nucleotide exchange factor (GEF) SOS1 domain. Structure-activity relationship studies on Rasarfin identify functionally selective analogs that inhibit AT1R endocytosis and/or MAPK activation. This project has also allowed for the development of novel biosensors that measure receptor-mediated signaling events in live cells. This study is the first to report a small molecule inhibitor of both Ras and Arf6 that can be further optimized for the functional characterization of Ras-mediated MAPK signaling versus Arf6-mediated receptor internalization in normal and cancer cells"--
Download or read book Regulation of G Protein Coupled Receptor Function and Expression written by Jeffrey L. Benovic and published by Wiley-Liss. This book was released on 1999-11-12 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recent advances in molecular and cell biology enabling the cloning, expression, and mutagenesis of signal transduction proteins has prompted an explosion of knowledge in the field of receptor regulation, facilitating the discovery of new classes of regulatory proteins, and providing a basis and means for manipulating receptor function through multiple intracellular targets. This volume covers methods used to examine how the function(s) of receptors are regulated. Understanding how to regulate the function and expression of these receptors is critical in determining how to modify receptors and to translocating receptors away from the cell surface and its recycling. Individual chapters focus on specific techniques used to characterize receptors (epitope tagging, measurement and analysis of receptor phosphorylation, analysis of the kinetics of receptor desensitization, and assessment of receptor/G protein coupling); the role of regulatory proteins (receptor kinases and phosphatases, arrestins) in modulating receptor function; and the methods used to measure receptor trafficking (ligand binding, immunofluoresence) and expression (transcriptional and translational regulation). * Covers a broad range of important concepts and methodologies which are current in the study of G protein-coupled receptors (GPCRs) * G-protein coupled receptors make up over 40% of the current pharmacological targets * Provides detailed protocols for executing various strategies and offers informed judgments as to what approaches are and aren't useful * Volume Editor, Jeffrey Benovic, is a dominant world leader in the study of receptor regulation of GPCR kinases and is highly respected in the field
Download or read book Receptor Receptor Interactions written by Kjell Fuxe and published by Springer Science & Business Media. This book was released on 2013-03-13 with total page 577 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Trafficking Inside Cells written by Nava Segev and published by Springer Science & Business Media. This book was released on 2010-05-30 with total page 459 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book covers the past, present and future of the intra-cellular trafficking field, which has made a quantum leap in the last few decades. It details how the field has developed and evolved as well as examines future directions.
Download or read book Functional Characterization of Protease activated Receptor 1 Palmitoylation in Receptor Signaling and Trafficking written by Isabel Canto Cordova and published by . This book was released on 2014 with total page 172 pages. Available in PDF, EPUB and Kindle. Book excerpt: G protein-coupled receptors (GPCRs) are the largest family of signaling receptors that respond to diverse stimuli and regulate many physiological responses. GPCRs elicit their cellular responses by coupling to distinct subtypes of heterotrimeric G-proteins composed of G[alpha] and G[beta][gamma] subunits. Activated GPCRs undergo conformational changes that allow the receptor to exchange GDP for GTP on the G[alpha] subunit, which induces dissociation from the [beta][gamma] subunits and subsequent downstream signaling. Protease-activated receptor-1 (PAR1) is a member of a family of GPCRs that are proteolytically activated. PAR1 is a receptor for the coagulant protease thrombin, and is capable of coupling to multiple G-protein subtypes to elicit various cellular responses; however, the mechanisms that regulate this selectivity are not well understood. Palmitoylation is a post-translational modification that many GPCRs possess, and is defined as the addition of palmitate, a 16 carbon fatty acid, to a cysteine residue via a thioester linkage. Many GPCRs are palmitoylated on their C-terminal tails, but the role of this modification differs based on the GPCR being examined. In this dissertation, I examined the role of palmitoylation in PAR1 signaling and trafficking. I defined the sites of PAR1 palmitoylation to occur on conserved C-tail cysteine residues C387 and C388. I discovered that palmitoylation is important for other PAR1 post-translational modifications, specifically phosphorylation and ubiquitination. I also show that palmitoylation of PAR1 regulates the accessibility of a nearby tyrosine-based sorting motif to the adaptor-protein complex-2 (AP-2) and -3 (AP-3), which controls receptor internalization and degradation. Additionally, palmitoylation appears to be important for the regulation of selective PAR1-induced signaling pathways such as G[alpha]12/13 -induced RhoA activation, G[alpha]i coupling, and thrombin-stimulated p38 MAPK signaling pathways. However, thrombin-induced G[alpha]q-mediated phosphoinositide hydrolysis and ERK1/2 activation are unperturbed in the absence of PAR1 palmitoylation. Taken together, the studies summarized in this dissertation highlight the relevance of palmitoylation for PAR1 function, and suggest that palmitoylation governs a C-tail conformation that is important for accessibility of other proteins such as ligases, kinases, adaptor proteins, and G-proteins, ultimately regulating PAR1 signaling and trafficking.
Download or read book G Protein Signaling Pathways in Health and Disease written by and published by Academic Press. This book was released on 2019-02-15 with total page 230 pages. Available in PDF, EPUB and Kindle. Book excerpt: G Protein Signaling Pathways in Health and Disease, Volume 161 in the Progress in Molecular Biology and Translational Science series, provides informative and exciting monographs on a wide variety of research topics related to G Protein Signaling Pathways in Health and Disease. The series gives in-depth knowledge on the important molecular biological aspects of organismal physiology and function, along with insights on how this knowledge can be applied to understand and ameliorate human disease. This updated release covers Diseases associated with mutations in CNGA3, Mutations in arrestins, Diseases caused by mutations in GPR101, Diseases caused by mutations in lutropin receptor, and much more. Comprises 15-20 chapters, providing substantial coverage on a given topic Contains ample use of tables, diagrams, schemata and color figures to enhance the reader's ability to rapidly grasp the information provided in each chapter Provides a comprehensive guide to the latest information available on prions, viruses, bacteria, and eukaryotes
