Methods In Enzymology, Volume 344

Section I: Activation of G Proteins by Receptors or Other Regulators [1]: Analysis of G Protein Activation in Sf9 and Mammalian Cells by Agonist-Promoted [35S]GTPgammaS Binding [2]: Elucidating Kinetic and Thermodynamic Constants for Interaction of G Protein Subunits and Receptors by Surface Plasmon Resonance Spectroscopy [3]: Neuroanatomical Localization of Receptor-Activated G Proteins in Brain [4]: Design and Use of C-Terminal Minigene Vectors for Studying Role of Heterotrimeric G Proteins [5]: Dissecting Receptor-G Protein Specificity Using Galpha Chimeras [6]: Use of Dominant Negative Mutations in Analysis of G Protein Function in Saccharomyces cerevisiae [7]: Functional Assays for Mammalian G-Protein-Coupled Receptors in Yeast [8]: Role of G Protein ssgamma Complex in Receptor-G Protein Interaction [9]: Phosducin Down-Regulation of G-Protein Coupling: Reconstitution of Phosducin Transducin of cGMP Cascade in Bovine Rod Photoreceptor Cells [10]: Analysis of Signal Transfer from Receptor to Go/Gi in Different Membrane Environments and Receptor-Independent Activators of Brain G Protein [11]: Identification of Modulators of Mammalian G-Protein Signaling by Functional Screens in the Yeast Saccharomyces cerevisiae Section II: Isolation or Production of Native or Modified [12]: Expression of alpha Subunit of Gs in Escherichia coli [13]: Purification of G Protein Isoforms GOA GOC from Bovine Brain [14]: Coexpression of Proteins with Methionine Aminopeptidase/or N-Myristoyltransferase in Escherichia coli to Increase Acylation Homogeneity of Protein Preparations [15]: Purification of G Protein ssgamma from Bovine Brain [16]: Separation and Analysis of G Protein gamma Subunits [17]: Activity of Ggamma Prenylcysteine Carboxyl Methyltransferase [18]: Preparation and Application of G Protein gamma Subunit-Derived Peptides Incorporating a Photoactive Isoprenoid Section III: Functional Analysis of G Protein Subunits [19]: Expression and Functional Analysis of G Protein alpha Subunits in S49 Lymphoma Cells [20]: Mouse Gene Knockout Knockin Strategies in Application to alpha Subunits of Gi/Go Family of G Proteins [21]: Determining Cellular Role of Galpha12 [22]: Targeted, Regulatable Expression of Activated Heterotrimeric G Protein alpha Subunits in Transgenic Mice [23]: Inducible, Tissue-Specific Suppression of Heterotrimeric G Protein alpha Subunits in Vivo [24]: Construction of Replication Defective Adenovirus That Expresses Mutant Galphas Q227L [25]: Expression of Adenovirus-Directed Expression of Activated Galphas in Rat Hippocampal Slices [26]: Quench-Flow Kinetic Measurement of Individual Reactions of G-Protein-Catalyzed GTPase Cycle [27]: Analysis of Genomic Imprinting of Gsalpha Gene [28]: Subcellular Localization of G Protein Subunits [29]: Fluorescence Approaches to Study G Protein Mechanisms [30]: Defining G Protein ssgamma Specificity for Effector Recognition [31]: Ribozyme-Mediated Suppression of G Protein gamma Subunits Section IV: G Protein Structure and Identification [32]: Use of Scanning Mutagenesis to Delineate Structure-Function Relationships in G Protein alpha Subunits [33]: Development of Gs-Selective Inhibitory Compounds [34]: Characterization of Deamidated G Protein Subunits [35]: Determining G Protein Heterotrimer Formation [36]: Use of Peptide Probes to Determine Function of Interaction Sites in G Protein Interactions with Effectors [37]: Protein Interaction Assays with G Proteins [38]: Evolutionary Traces of Functional Surfaces along G Protein Signaling Pathway [39]: Discovery of Ligands for ssgamma Subunits from Phage-Displayed Peptide Libraries [40]: Exploring Protein-Protein Interactions by Peptide Docking Protocols [41]: Structural Characterization of Intact G Protein gamma Subunits by Mass Spectrometry Section V: RGS Proteins and Signal Termination [42]: Quantitative Assays for GTPase-Activating Proteins [43]: Analysis of RGS Proteins in Saccharomyces cerevisiae [44]: Purification of RGS Protein, Sst2, from Saccharomyces cerevisiae and Escherichia coli [45]: RGS Domain: Production and Uses of Recombinant Protein [46]: Screening for Interacting Partners for Galphai3 and RGS-GAIP Using the Two-Hybrid System [47]: Assay of RGS Protein Activity in Vitro Using Purified Components [48]: Measuring RGS Protein Interactions with Gqalpha [49]: Assays of Complex Formation between RGS Protein Ggamma Subunit-like Domains and Gss Subunits [50]: RGS Function in Visual Signal Transduction [51]: Molecular Cloning of Regulators of G-Protein Signaling Family Members and Characterization of Binding Specificity of RGS 12 PDZ Domain Author index Subject Index