G Protein Pathways, Part B: G Proteins and Their Regulators

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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. The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today—truly an essential publication for researchers in all fields of life sciences.

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]GTP?S 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 Ga 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 ß? 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 a 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 ß? from Bovine Brain

[16]: Separation and Analysis of G Protein ? Subunits

[17]: Activity of G? Prenylcysteine Carboxyl Methyltransferase

[18]: Preparation and Application of G Protein ? Subunit-Derived Peptides Incorporating a Photoactive Isoprenoid

Section III: Functional Analysis of G Protein Subunits

[19]: Expression and Functional Analysis of G Protein a Subunits in S49 Lymphoma Cells

[20]: Mouse Gene Knockout Knockin Strategies in Application to a Subunits of Gi/Go Family of G Proteins

[21]: Determining Cellular Role of Ga12

[22]: Targeted, Regulatable Expression of Activated Heterotrimeric G Protein a Subunits in Transgenic Mice

[23]: Inducible, Tissue-Specific Suppression of Heterotrimeric G Protein a Subunits in Vivo

[24]: Construction of Replication Defective Adenovirus That Expresses Mutant Gas Q227L

[25]: Expression of Adenovirus-Directed Expression of Activated Gas in Rat Hippocampal Slices

[26]: Quench-Flow Kinetic Measurement of Individual Reactions of G-Protein-Catalyzed GTPase Cycle

[27]: Analysis of Genomic Imprinting of Gsa Gene

[28]: Subcellular Localization of G Protein Subunits

[29]: Fluorescence Approaches to Study G Protein Mechanisms

[30]: Defining G Protein ß? Specificity for Effector Recognition

[31]: Ribozyme-Mediated Suppression of G Protein ? Subunits

Section IV: G Protein Structure and Identification

[32]: Use of Scanning Mutagenesis to Delineate Structure–Function Relationships in G Protein a 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 ß? Subunits from Phage-Displayed Peptide Libraries

[40]: Exploring Protein–Protein Interactions by Peptide Docking Protocols

[41]: Structural Characterization of Intact G Protein ? 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 Gai3 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 Gqa

[49]: Assays of Complex Formation between RGS Protein G? 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

Biochemists, Molecular Biologists, Cell Biologists, Pharmacologists, Neurophysiologists, Neurochemists, Neuroendocrinologists, and Biomedical Researchers.