G protein signaling : methods and protocols / edited by Alan V. Smrcka.

Contributor(s): Smrcka, Alan V
Material type: TextTextSeries: Methods in molecular biology (Clifton, N.J.): v. 237.Publisher: Totowa, NJ : Humana Press, ©2004Description: 1 online resource (xii, 249 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceISBN: 9781592594306; 1592594301; 1588291375; 9781588291370Subject(s): G proteins -- Laboratory manuals | Cellular signal transduction -- Laboratory manuals | GTP-Binding Proteins -- physiology | Genetic Techniques | GTP-Binding Protein Regulators -- physiology | Clinical Laboratory Techniques -- methods | SCIENCE -- Life Sciences -- Biochemistry | Cellular signal transduction | G proteinsGenre/Form: Laboratory Manual. | Electronic books. | Handbooks and manuals. Additional physical formats: Print version:: G protein signaling.DDC classification: 572/.643 LOC classification: QP552.G16 | G185 2004Online resources: Click here to access online
Contents:
Purification of recombinant g protein alpha subunits from escherichia coli / W.K. Greentree and M.E. Linder -- Purification of g protein subunits from sf9 insect cells using hexahistidine-tagged alpha and beta gamma subunits / T. Kozasa -- Expression and purification of soluble adenylyl cyclase from escherichia coli / J.A. Beeler and W.J. Tang -- Purification of phospholipase c beta and phospholipase c epsilon from sf9 cells / M. Ghosh [and others] -- Assay for g protein-dependent activation of phospholipase c beta using purified protein components / M. Ghosh and A.V. Smrcka -- Assays of recombinant adenylyl cyclases expressed in sf9 cells / R. Taussig -- Measurement of g protein-coupled receptor-stimulated phospholipase d activity in intact cells / S.J. Walker and H.A. Brown -- Analysis of g protein-mediated activation of phospholipase c in cultured cells / B. Liu and D. Wu -- Intensive mutational analysis of g protein-coupled receptors in yeast / A. Celic [and others] -- Green fluorescent protein-tagged beta-arrestin translocation as a measure of g protein-coupled receptor activation / S.S. Ferguson and M.G. Caron -- Detection of g protein-coupled receptors by immunofluorescence microscopy / P.M. Hinkle and J.A. Puskas -- Assay of g protein-coupled receptor activation of g proteins in native cell membranes using [35s]gtp gamma s binding / J.M. Bidlack and A.L. Parkhill -- Analysis of the coupling of g12/13 to g protein-coupled receptors using a luciferase reporter assay / B. Liu and D. Wu -- Selective inhibition of g protein-mediated pathways using rgs domains / T. Kozasa and R.D. Ye -- Ribozymes as tools for suppression of g protein gamma subunits / J.D. Robishaw, Z.P. Guo and Q. Wang -- In vivo adenoviral-mediated gene transfer of the beta arkct to study the role of g beta gamma in arterial restenosis / G. Iaccarino and W.J. Koch -- Analysis of rgs protein palmitoylation / L.S. Bernstein, M.E. Linder and J.R. Hepler -- Methods for measuring rgs protein phosphorylation by g protein-regulated kinases / S. Hollinger and J.R. Hepler -- The use of green fluorescent proteins to view association between phospholipase c beta and g protein subunits in cells / S. Scarlata and L. Dowal -- Cellular localization of gfp-tagged alpha subunits / T.R. Hynes, T.E. Hughes and C.H. Berlot.
In: Springer ProtocolsSummary: G 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.
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Includes bibliographical references and index.

Purification of recombinant g protein alpha subunits from escherichia coli / W.K. Greentree and M.E. Linder -- Purification of g protein subunits from sf9 insect cells using hexahistidine-tagged alpha and beta gamma subunits / T. Kozasa -- Expression and purification of soluble adenylyl cyclase from escherichia coli / J.A. Beeler and W.J. Tang -- Purification of phospholipase c beta and phospholipase c epsilon from sf9 cells / M. Ghosh [and others] -- Assay for g protein-dependent activation of phospholipase c beta using purified protein components / M. Ghosh and A.V. Smrcka -- Assays of recombinant adenylyl cyclases expressed in sf9 cells / R. Taussig -- Measurement of g protein-coupled receptor-stimulated phospholipase d activity in intact cells / S.J. Walker and H.A. Brown -- Analysis of g protein-mediated activation of phospholipase c in cultured cells / B. Liu and D. Wu -- Intensive mutational analysis of g protein-coupled receptors in yeast / A. Celic [and others] -- Green fluorescent protein-tagged beta-arrestin translocation as a measure of g protein-coupled receptor activation / S.S. Ferguson and M.G. Caron -- Detection of g protein-coupled receptors by immunofluorescence microscopy / P.M. Hinkle and J.A. Puskas -- Assay of g protein-coupled receptor activation of g proteins in native cell membranes using [35s]gtp gamma s binding / J.M. Bidlack and A.L. Parkhill -- Analysis of the coupling of g12/13 to g protein-coupled receptors using a luciferase reporter assay / B. Liu and D. Wu -- Selective inhibition of g protein-mediated pathways using rgs domains / T. Kozasa and R.D. Ye -- Ribozymes as tools for suppression of g protein gamma subunits / J.D. Robishaw, Z.P. Guo and Q. Wang -- In vivo adenoviral-mediated gene transfer of the beta arkct to study the role of g beta gamma in arterial restenosis / G. Iaccarino and W.J. Koch -- Analysis of rgs protein palmitoylation / L.S. Bernstein, M.E. Linder and J.R. Hepler -- Methods for measuring rgs protein phosphorylation by g protein-regulated kinases / S. Hollinger and J.R. Hepler -- The use of green fluorescent proteins to view association between phospholipase c beta and g protein subunits in cells / S. Scarlata and L. Dowal -- Cellular localization of gfp-tagged alpha subunits / T.R. Hynes, T.E. Hughes and C.H. Berlot.

Print version record.

G 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.

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