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.

Table of 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.