G Protein Signaling

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 ŽCeliŽc, 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