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020 _a9781592594306
024 7 _a10.1385/1592594301
_2doi
040 _aES-MaUEC
_bspa
_cES-MaUEC
245 1 0 _aG Protein Signaling
_bMethods and Protocols
_cedited by Alan V. Smrcka.
250 _a1st edition 2004
264 1 _aTotowa, NJ
_bHumana Press
_c2004
300 _a1 recurso en línea (XII, 250 páginas)
_b
336 _atexto
_btxt
_2rdacontent
337 _aelectrónico
_bc
_2rdamedia
338 _arecurso electrónico
_bcr
_2rdacarrier
347 _aarchivo de texto
_bPDF
490 0 _aMethods in Molecular Biology
_x1940-6029
_v237
505 0 _aPurification of G Protein Subunits and G Protein Effectors -- Purification of Recombinant G Protein ? Subunits from Escherichia coli -- Purification of G Protein Subunits from Sf9 Insect Cells Using Hexahistidine-Tagged ? and ?? Subunits -- Expression and Purification of Soluble Adenylyl Cyclase from Escherichia coli -- Purification of Phospholipase C ? and Phospholipase C ? from Sf9 Cells -- Assays for G Protein-Dependent Effector Activation In Vitro -- Assay for G Protein-Dependent Activation of Phospholipase C ? Using Purified Protein Components -- Assays of Recombinant Adenylyl Cyclases Expressed in Sf9 Cells -- Assays For G Protein-Coupled Receptor-Mediated and G Protein Subunit-Mediated Effector Activation in Intact Cells -- Measurement of G Protein-Coupled Receptor-Stimulated Phospholipase D Activity in Intact Cells -- Analysis of G Protein-Mediated Activation of Phospholipase C in Cultured Cells -- Receptor Mechanisms, Activity, and Localization -- Intensive Mutational Analysis of G Protein-Coupled Receptors in Yeast -- Green Fluorescent Protein-Tagged ?-Arrestin Translocation as a Measure of G Protein-Coupled Receptor Activation -- Detection of G Protein-Coupled Receptors by Immunofluorescence Microscopy -- Assay of G Protein-Coupled Receptor Activation of G Proteins in Native Cell Membranes Using [35S]GTP?S Binding -- Analysis of the Coupling of G12/13 to G Protein-Coupled Receptors Using a Luciferase Reporter Assay -- Disruption of Endogenous G Protein-Signaling Systems to Understand the Role of G Protein Subunits in Intact Systems -- Selective Inhibition of G Protein-Mediated Pathways Using RGS Domains -- Ribozymes as Tools for Suppression of G Protein ? Subunits -- In Vivo Adenoviral-Mediated Gene Transfer of the ?ARKct to Study the Role of G?? in Arterial Restenosis -- Posttranslational Modifications of Regulators of G Protein Signaling -- Analysis of RGS Protein Palmitoylation -- Methods for Measuring RGS Protein Phosphorylation by G Protein-Regulated Kinases -- Use of Gfp-Tagged G Proteins and Effectors in Intact Cells -- The Use of Green Fluorescent Proteins to View Association Between Phospholipase C? and G Protein Subunits in Cells -- Cellular Localization of GFP-Tagged ? Subunits.
520 _aG proteins and G protein-coupled receptors are ubiquitously expressed proteins that regulate a wide range of physiological processes and are consequently the targets of many pharmaceuticals. In G Protein Signaling: Methods and Protocols, leading researchers describe in detail key methods for investigating G protein signaling from a variety of perspectives ranging from in vitro biochemistry to whole animal studies. The authors focus on the mechanisms of G protein and G protein-coupled receptor function and the roles of G protein subunits in cell biology and disease. Among the readily reproducible techniques presented are those for the purification of G proteins and effector enzymes, assays of these purified G proteins and effector enzymes, and for the study of G protein interactions with effectors in intact cells. Additional methods are provided for assaying G protein-coupled receptor structure, function, and localization, for studying the physiological roles for endogenous G proteins, and for examining lipid and phosphate modifications of RGS proteins. Each fully tested protocol includes a background introduction explaining the principle behind the technique, equipment and reagent lists, tips on troubleshooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results. Comprehensive and highly practical, G Protein Signaling: Methods and Protocols offers novice and expert researchers alike an array of cutting-edge tools to illuminate the roles of G protein and G protein-coupled receptors, as well as a better understanding of their action with drugs, neurotransmitters, and sensory stimuli.
700 1 _aSmrcka, Alan V
_eeditor literario
_4edt
_4http://id.loc.gov/vocabulary/relators/edt
776 0 8 _iPrinted edition:
_z9781617373817
776 0 8 _iPrinted edition:
_z9781588291370
776 0 8 _iPrinted edition:
_z9781489939418
856 4 0 _uhttps://go.openathens.net/redirector/universidadeuropea.es?url=https://doi.org/10.1385/1592594301
_z(usuarios Universidad Europea de Valencia)
942 _2lcc
_cLE
988 _aSpringer_Protocols_2004
999 _c235592
_d235592