G Protein Signaling: Methods and Protocols [Hardcover]

Purification 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 ProteinDependent 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 ArterialRestenosis.- 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-Tal³æ