G Protein Signaling: Methods and Protocols (Paperback)

Part I. Purification of G Protein Subunits and G Protein Effectors Purification of Recombinant G Protein a Subunits from Escherichia coli Wendy K. Greentree and Maurine E. Linder Purification of G Protein Subunits from Sf9 Insect Cells Using Hexahistidine-Tagged a and bg Subunits Tohru Kozasa Expression and Purification of Soluble Adenylyl Cyclase from Eschericia coli Jeff A. Beeler and Wei-Jen Tang Purification of Phospholipase C b and Phospholipase C e from Sf9 Cells Mousumi Ghosh, Huan Wang, Grant G. Kelley, and Alan V. Smrcka Part II. Assays for G Protein-Dependent Effector Activation In Vitro Assay for G Protein-Dependent Activation of Phospholipase C b Using Purified Protein Components Mousumi Ghosh and Alan V. Smrcka Assays of Recombinant Adenylyl Cyclases Expressed in Sf9 Cells Ronald Taussig Part III. 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 Stephanie J. Walker and H. Alex Brown Analysis of G Protein-Mediated Activation of Phospholipase C in Cultured Cells Bo Liu and Dianqing Wu Part IV. Receptor Mechanisms, Activity, and Localization Intensive Mutational Analysis of G Protein-Coupled Receptors in Yeast Andjelka Celi c, Sara M. Connelly, Negin P. Martin, and Mark E. Dumont Green Fluorescent Protein-Tagged b-Arrestin Translocation as a Measure of G Protein-Coupled Receptor Activation Stephen S. G. Ferguson and Marc G. Caron Detection of G Protein-Coupled Receptors by Immunofluorescence Microscopy Patricia M. Hinkle and John A. Puskas Assay of G Protein-Coupled Receptor Activation of G Proteins in Native Cell Membranes Using [35S]GTPgS Binding Jean M. Bidlack and Amy L.Parkhill Analysis of the Coupling of G12/13 to G Protein-Coupled Receptors Using a Luciferase Reporter Assay Bo Liu and Dianqing Wu Part V. 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 Tohru Kozasa and Richard D. Ye Ribozymes as Tools for Suppression of G Protein g Subunits Janet D. Robishaw, Zheng-ping Guo, and Qin Wang In Vivo Adenoviral-Mediated Gene Transfer of the bARKct to Study the Role of Gbg in Arterial Restenosis Guido Iaccarino and Walter J. Koch Part VI. Posttranslational Modifications of Regulators of G Protein Signaling Analysis of RGS Protein Palmitoylation Leah S. Bernstein, Maurine E. Linder, and John R. Hepler Methods for Measuring RGS Protein Phosphorylation by G Protein-Regulated Kinases Susanne Hollinger and John R. Hepler Part VII. Use of GFP-Tagged G Proteins and Effectors in Intact Cells The Use of Green Fluorescent Proteins to View Association Between Phospholipase Cb and G Protein Subunits in Cells Suzanne Scarlata and Louisa Dowal Cellular Localization of GFP-Tagged a Subunits Thomas R. Hynes, Thomas E. Hughes, and Catherine H. Berlot Index