G Protein Pathways, Part C: Effector Mechanisms book cover

G Protein Pathways, Part C: Effector Mechanisms

This third volume in the trio covering G proteins, features integrated approaches to studying G proteins. Methods pertaining to signaling mechanisms are presented, including theoretical and modeling approaches, biochemistry and molecular biology, and cell biology and physiology. The techniques for studying the structure and function of G proteins are important not only to those with specific research interests in them, but also endocrinologists and pharmacologists conducting research on signaling mechanisms that are increasingly understood to interact with G proteins.

Audience
Biochemists, molecular biologists, cell biologists, pharmacologists, neurophysiologists, neurochemists, neuroendocrinologists, and biomedical researchers.

Included in series
Methods in Enzymology

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Published: October 2001

Imprint: Academic Press

ISBN: 978-0-12-182246-0

Contents

  • Contributors to Volume 345.Preface.Volume in Series.Section I: Modeling Intracellular Signaling Pathways:Use of Kinetikit and GENESIS for Modeling Signaling Pathways.Section II: Phosphodiesterases:Assays of G Protein/cGMP-Phosphodiesterase Interactions.Assaying G Protein-Phosphodiesterase Interactions in Sensory Systems.Section III: Calcium and Potassium Channels:Studies of Endogenous G-Protein-Mediated Pathways in Neurons by Whole-Cell Electrophysiology.Biochemical Approaches to Study Interaction of Calcium Channels with RGS12 in Primary Neuronal Cultures.Assaying Phosphatidylinositol Bisphosphate Regulation of Potassium Channels.Section IV: Adenylyl Cyclases:Purification of Soluble Adenylyl Cyclase.Calcium-Sensitive Adenylyl Cyclase/Aequorin Chimeras as Sensitive Probes for Discrete Modes of Elevation of Cytosolic Calcium.Kinetic Analysis of the Action of P-Site Analogs.Expression, Purification, and Assay of Cytosolic (Catalytic) Domains of Membrane-Bound Mammalian Adenylyl Cyclases.Identification of Putative Direct Effectors for Ga0 Using Yeast Two-Hybrid Method.Identification of Transmembrane Adenylyl Cyclase Isoforms.Functional Analyses of Type V Adenylyl Cyclase.Photoaffinity Labeling of Adenylyl Cyclase.Crystallization of Complex between Soluble Domains of Adenylyl Cyclase and Activated Gsa.Generation of Adenylyl Cyclase Knockout Mice.Construction of Soluble Adenylyl Cyclase from Human Membrane-Bound Type 7 Adenylyl Cyclase.Section V: Phospholipases and Lipid-Derived Products:Expression and Characterization of Rat Brain Phospholipase D.G-Protein-Coupled Receptor Regulation of Phospholipase D.Analysis and Quantitation of Ceramide.Assays for Phospholipase D Reaction Products.Determination of Strength and Specificity of Membrane-Bound G Protein-Phospholipase C Association Using Fluorescence Spectroscopy.Assays and Characterization of Mammalian Phosphatidylinositol 4,5-Bisphosphate-Sensitive Phospholipase D.Characterization and Purification of Phosphatidylinositol Trisphosphate 5-Phosphatase from Rat Brain Tissues.Section VI: Small GTP-Binding Proteins:Assay of Cdc42, Rac, and Rho GTPase Activation by Affinity Methods.Assays of ADP-Ribosylation Factor Function.Functional Characterization of p115 RhoGEF.Nonisotopic Methods for Detecting Activation of Small G Proteins.BIG1 and BIG2: Brefeldin A-Inhibited Guanine Nucleotide-Exchange Proteins for ADP-Ribosylation Factors.Functional Interaction of Ga13 with p115RhoGEF Determined with Transcriptional Reporter System.Section VII: Protein Kinases and Phosphatases:Analysis of c-Jun N-Terminal Kinase Regulation and Function.Double-Label Confocal Microscopy of Phosphorylated Protein Kinases Involved in Long-Term Potentiation.Regulation of Mitogen-Activated Protein Kinases by G-Protein-Coupled Receptors.Analysis of Protein Kinase B/Akt.Direct Stimulation of Bruton's Tyrosine Kinase by G Protein a Subunits.Isozyme-Specific Inhibitors and Activators of Protein Kinase C.Assay of Raf-1 Activity.Analyzing Protein Kinase C. Activation.Assays for Protein-Tyrosine Phosphates.Differential Display of mRNAs Regulated by G-Protein Signaling.Gene Profiling of Transgenic Mice with Targeted Expression of Activated Heterotrimeric G Protein a Subunits Using DNA Microarray.Retroviral Vectors Applied to Gene Regulation Studies.Overexpression of Tightly Regulated Proteins: Protein Phosphatase 2A Overexpression in NIH 3T3 Cells.Monitoring G-Protein-Coupled Receptor Signaling with DNA Microarrays and Real-Time Polymerase Chain Reaction.CAMP Response Element-Mediated Gene Expression in Transgenic Reporter Gene Mouse Strain.Functional Genomic Search of G-Protein-Coupled Receptors Using Microarrays with Normalized cDNA Library.Author Index.Subject Index.

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