This volume covers topics such as the structure and identification of functional domains of G proteins, and activation of G proteins by receptors or other regulators. The text takes an integrated approach to studying common experimental questions at many different levels related to G proteins. Methods related to G proteins using molecular modeling, systems biology, protein engineering, protein biochemistry, cell biology, and physiology are all accessible in the same volume.
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Section I: Activation of G Proteins by Receptors or Other Regulators : Analysis of G Protein Activation in Sf9 and Mammalian Cells by Agonist-Promoted [35S]GTPgammaS Binding : Elucidating Kinetic and Thermodynamic Constants for Interaction of G Protein Subunits and Receptors by Surface Plasmon Resonance Spectroscopy : Neuroanatomical Localization of Receptor-Activated G Proteins in Brain : Design and Use of C-Terminal Minigene Vectors for Studying Role of Heterotrimeric G Proteins : Dissecting Receptor-G Protein Specificity Using Galpha Chimeras : Use of Dominant Negative Mutations in Analysis of G Protein Function in Saccharomyces cerevisiae : Functional Assays for Mammalian G-Protein-Coupled Receptors in Yeast : Role of G Protein ssgamma Complex in Receptor-G Protein Interaction : Phosducin Down-Regulation of G-Protein Coupling: Reconstitution of Phosducin Transducin of cGMP Cascade in Bovine Rod Photoreceptor Cells : Analysis of Signal Transfer from Receptor to Go/Gi in Different Membrane Environments and Receptor-Independent Activators of Brain G Protein : 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 : Expression of alpha Subunit of Gs in Escherichia coli : Purification of G Protein Isoforms GOA GOC from Bovine Brain : Coexpression of Proteins with Methionine Aminopeptidase/or N-Myristoyltransferase in Escherichia coli to Increase Acylation Homogeneity of Protein Preparations : Purification of G Protein ssgamma from Bovine Brain : Separation and Analysis of G Protein gamma Subunits : Activity of Ggamma Prenylcysteine Carboxyl Methyltransferase : Preparation and Application of G Protein gamma Subunit-Derived Peptides Incorporating a Photoactive Isoprenoid Section III: Functional Analysis of G Protein Subunits : Expression and Functional Analysis of G Protein alpha Subunits in S49 Lymphoma Cells : Mouse Gene Knockout Knockin Strategies in Application to alpha Subunits of Gi/Go Family of G Proteins : Determining Cellular Role of Galpha12 : Targeted, Regulatable Expression of Activated Heterotrimeric G Protein alpha Subunits in Transgenic Mice : Inducible, Tissue-Specific Suppression of Heterotrimeric G Protein alpha Subunits in Vivo : Construction of Replication Defective Adenovirus That Expresses Mutant Galphas Q227L : Expression of Adenovirus-Directed Expression of Activated Galphas in Rat Hippocampal Slices : Quench-Flow Kinetic Measurement of Individual Reactions of G-Protein-Catalyzed GTPase Cycle : Analysis of Genomic Imprinting of Gsalpha Gene : Subcellular Localization of G Protein Subunits : Fluorescence Approaches to Study G Protein Mechanisms : Defining G Protein ssgamma Specificity for Effector Recognition : Ribozyme-Mediated Suppression of G Protein gamma Subunits Section IV: G Protein Structure and Identification : Use of Scanning Mutagenesis to Delineate Structure-Function Relationships in G Protein alpha Subunits : Development of Gs-Selective Inhibitory Compounds : Characterization of Deamidated G Protein Subunits : Determining G Protein Heterotrimer Formation : Use of Peptide Probes to Determine Function of Interaction Sites in G Protein Interactions with Effectors : Protein Interaction Assays with G Proteins : Evolutionary Traces of Functional Surfaces along G Protein Signaling Pathway : Discovery of Ligands for ssgamma Subunits from Phage-Displayed Peptide Libraries : Exploring Protein-Protein Interactions by Peptide Docking Protocols : Structural Characterization of Intact G Protein gamma Subunits by Mass Spectrometry Section V: RGS Proteins and Signal Termination : Quantitative Assays for GTPase-Activating Proteins : Analysis of RGS Proteins in Saccharomyces cerevisiae : Purification of RGS Protein, Sst2, from Saccharomyces cerevisiae and Escherichia coli : RGS Domain: Production and Uses of Recombinant Protein : Screening for Interacting Partners for Galphai3 and RGS-GAIP Using the Two-Hybrid System : Assay of RGS Protein Activity in Vitro Using Purified Components : Measuring RGS Protein Interactions with Gqalpha : Assays of Complex Formation between RGS Protein Ggamma Subunit-like Domains and Gss Subunits : RGS Function in Visual Signal Transduction : Molecular Cloning of Regulators of G-Protein Signaling Family Members and Characterization of Binding Specificity of RGS 12 PDZ Domain Author index Subject Index
Series: Methods in Enzymology : Book 344
Number Of Pages: 813
Published: 11th December 2001
Publisher: Elsevier Science Publishing Co Inc
Country of Publication: US
Dimensions (cm): 22.9 x 15.2
Weight (kg): 1.23