PRAISE FOR THE SERIES"The Methods in Enzymology series represents the gold-standard." --NEUROSCIENCE"Incomparably useful." --ANALYTICAL BIOCHEMISTRY"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." --BIO/TECHNOLOGY"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." --AMERICAN SOCIETY OF MICROBIOLOGY NEWS"If we had some way to find the work most often consulted in the laboratory, it could well be Colowick and Kaplan's multi-volume series Methods in Enzymology...a great work." --ENZYMOLOGIA"A series that has established itself as a definitive reference for biochemists." --JOURNAL OF CHROMATOGRAPHY

• 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]GTPγS 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 Gα 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 βγ 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 cerevisiaeSection II: Isolation or Production of Native or Modified[12]: Expression of α 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 βγ from Bovine Brain[16]: Separation and Analysis of G Protein γ Subunits[17]: Activity of Gγ Prenylcysteine Carboxyl Methyltransferase[18]: Preparation and Application of G Protein γ Subunit-Derived Peptides Incorporating a Photoactive IsoprenoidSection III: Functional Analysis of G Protein Subunits[19]: Expression and Functional Analysis of G Protein α Subunits in S49 Lymphoma Cells[20]: Mouse Gene Knockout Knockin Strategies in Application to α Subunits of Gi/Go Family of G Proteins[21]: Determining Cellular Role of Gα12[22]: Targeted, Regulatable Expression of Activated Heterotrimeric G Protein α Subunits in Transgenic Mice[23]: Inducible, Tissue-Specific Suppression of Heterotrimeric G Protein α Subunits in Vivo[24]: Construction of Replication Defective Adenovirus That Expresses Mutant Gαs Q227L[25]: Expression of Adenovirus-Directed Expression of Activated Gαs in Rat Hippocampal Slices[26]: Quench-Flow Kinetic Measurement of Individual Reactions of G-Protein-Catalyzed GTPase Cycle[27]: Analysis of Genomic Imprinting of Gsα Gene[28]: Subcellular Localization of G Protein Subunits[29]: Fluorescence Approaches to Study G Protein Mechanisms[30]: Defining G Protein βγ Specificity for Effector Recognition[31]: Ribozyme-Mediated Suppression of G Protein γ SubunitsSection IV: G Protein Structure and Identification[32]: Use of Scanning Mutagenesis to Delineate Structure–Function Relationships in G Protein α 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 βγ Subunits from Phage-Displayed Peptide Libraries[40]: Exploring Protein–Protein Interactions by Peptide Docking Protocols[41]: Structural Characterization of Intact G Protein γ Subunits by Mass SpectrometrySection 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 Gαi3 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 Gqα[49]: Assays of Complex Formation between RGS Protein Gγ Subunit-like Domains and Gβ 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 DomainAuthor indexSubject Index

PRAISE FOR THE SERIES"The Methods in Enzymology series represents the gold-standard." --NEUROSCIENCE"Incomparably useful." --ANALYTICAL BIOCHEMISTRY"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." --BIO/TECHNOLOGY"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." --AMERICAN SOCIETY OF MICROBIOLOGY NEWS"If we had some way to find the work most often consulted in the laboratory, it could well be Colowick and Kaplan's multi-volume series Methods in Enzymology...a great work." --ENZYMOLOGIA"A series that has established itself as a definitive reference for biochemists." --JOURNAL OF CHROMATOGRAPHY

• 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]GTPγS 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 Gα 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 βγ 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 cerevisiaeSection II: Isolation or Production of Native or Modified[12]: Expression of α 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 βγ from Bovine Brain[16]: Separation and Analysis of G Protein γ Subunits[17]: Activity of Gγ Prenylcysteine Carboxyl Methyltransferase[18]: Preparation and Application of G Protein γ Subunit-Derived Peptides Incorporating a Photoactive IsoprenoidSection III: Functional Analysis of G Protein Subunits[19]: Expression and Functional Analysis of G Protein α Subunits in S49 Lymphoma Cells[20]: Mouse Gene Knockout Knockin Strategies in Application to α Subunits of Gi/Go Family of G Proteins[21]: Determining Cellular Role of Gα12[22]: Targeted, Regulatable Expression of Activated Heterotrimeric G Protein α Subunits in Transgenic Mice[23]: Inducible, Tissue-Specific Suppression of Heterotrimeric G Protein α Subunits in Vivo[24]: Construction of Replication Defective Adenovirus That Expresses Mutant Gαs Q227L[25]: Expression of Adenovirus-Directed Expression of Activated Gαs in Rat Hippocampal Slices[26]: Quench-Flow Kinetic Measurement of Individual Reactions of G-Protein-Catalyzed GTPase Cycle[27]: Analysis of Genomic Imprinting of Gsα Gene[28]: Subcellular Localization of G Protein Subunits[29]: Fluorescence Approaches to Study G Protein Mechanisms[30]: Defining G Protein βγ Specificity for Effector Recognition[31]: Ribozyme-Mediated Suppression of G Protein γ SubunitsSection IV: G Protein Structure and Identification[32]: Use of Scanning Mutagenesis to Delineate Structure–Function Relationships in G Protein α 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 βγ Subunits from Phage-Displayed Peptide Libraries[40]: Exploring Protein–Protein Interactions by Peptide Docking Protocols[41]: Structural Characterization of Intact G Protein γ Subunits by Mass SpectrometrySection 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 Gαi3 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 Gqα[49]: Assays of Complex Formation between RGS Protein Gγ Subunit-like Domains and Gβ 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 DomainAuthor indexSubject Index

PRAISE FOR THE SERIES"The Methods in Enzymology series represents the gold-standard." --NEUROSCIENCE"Incomparably useful." --ANALYTICAL BIOCHEMISTRY"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." --BIO/TECHNOLOGY"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." --CHEMISTRY IN INDUSTRY"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." --AMERICAN SOCIETY OF MICROBIOLOGY NEWS"If we had some way to find the work most often consulted in the laboratory, it could well be Colowick and Kaplan's multi-volume series Methods in Enzymology...a great work." --ENZYMOLOGIA"A series that has established itself as a definitive reference for biochemists." --JOURNAL OF CHROMATOGRAPHY

• 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]GTPγS 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 Gα 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 βγ 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 cerevisiaeSection II: Isolation or Production of Native or Modified[12]: Expression of α 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 βγ from Bovine Brain[16]: Separation and Analysis of G Protein γ Subunits[17]: Activity of Gγ Prenylcysteine Carboxyl Methyltransferase[18]: Preparation and Application of G Protein γ Subunit-Derived Peptides Incorporating a Photoactive IsoprenoidSection III: Functional Analysis of G Protein Subunits[19]: Expression and Functional Analysis of G Protein α Subunits in S49 Lymphoma Cells[20]: Mouse Gene Knockout Knockin Strategies in Application to α Subunits of Gi/Go Family of G Proteins[21]: Determining Cellular Role of Gα12[22]: Targeted, Regulatable Expression of Activated Heterotrimeric G Protein α Subunits in Transgenic Mice[23]: Inducible, Tissue-Specific Suppression of Heterotrimeric G Protein α Subunits in Vivo[24]: Construction of Replication Defective Adenovirus That Expresses Mutant Gαs Q227L[25]: Expression of Adenovirus-Directed Expression of Activated Gαs in Rat Hippocampal Slices[26]: Quench-Flow Kinetic Measurement of Individual Reactions of G-Protein-Catalyzed GTPase Cycle[27]: Analysis of Genomic Imprinting of Gsα Gene[28]: Subcellular Localization of G Protein Subunits[29]: Fluorescence Approaches to Study G Protein Mechanisms[30]: Defining G Protein βγ Specificity for Effector Recognition[31]: Ribozyme-Mediated Suppression of G Protein γ SubunitsSection IV: G Protein Structure and Identification[32]: Use of Scanning Mutagenesis to Delineate Structure–Function Relationships in G Protein α 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 βγ Subunits from Phage-Displayed Peptide Libraries[40]: Exploring Protein–Protein Interactions by Peptide Docking Protocols[41]: Structural Characterization of Intact G Protein γ Subunits by Mass SpectrometrySection 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 Gαi3 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 Gqα[49]: Assays of Complex Formation between RGS Protein Gγ Subunit-like Domains and Gβ 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 DomainAuthor indexSubject Index