Overviews -- Peptide Recognition Mechanisms of Eukaryotic Signaling Modules -- Protein-Protein Interactions in Signaling Cascades -- Transmembrane Signaling by Receptor Oligomerization -- Specific Topics -- Use of Peptide Libraries to Determine Optimal Substrates of Tyrosine Kinases -- Mapping the Specificity of SH3 Domains with Phage-Displayed Random-Peptide Libraries -- Selective Antagonism of Receptor Signaling Using Antisense RNA to Deplete G-Protein Subunits -- Microinjection of Antisense Oligonucleotides and Electrophysiological Recording of Whole-Cell Currents as Tools to identify Specific G-Protein Subtypes Coupling Hormone Receptors to Voltage-Gated Calcium Channels -- Oocytes Microinjection Assay to Study the MAP-Kinase Cascade -- Mammalian Cell Microinjection Assay to Study the Function of Rac and Rho -- Identification and Functional Reconstitution of Effector Proteins for the GTPases Rac and CDC42HS -- Cell-Free Assay System for Ras- and Rap1-Dependent Activation of MAP-Kinase Cascade -- Cell-Free Assay System for Ras- and Rap1-Dependent Activation of MAP-Kinase Cascade -- Reconstitution System Based on Cytosol-Depleted Cells to Study the Regulation of Phospholipases C and D -- Reconstitution System Based on Cytosol-Depleted Cells to Study the Regulation of Phospholipases C and D -- Two-Hybrid Analysis of Ras-Raf Interactions -- Two-Hybrid Analysis of Ras-Raf Interactions -- Cloning and Mutational Analysis of the She-Phosphotyrosine Interaction/ Phosphotyrosine-Binding Domain -- Cloning and Mutational Analysis of the Shc-Phosphotyrosine Interaction/Phosphotyrosine-Binding Domain -- Use of Fluorescence Spectroscopy to Study the Regulation of Small G Proteins -- Use of Fluorescence Spectroscopy to Study the Regulation of Small G Proteins -- Prenylation Assays for Small GTPases -- Prenylation Assays for Small GTPases -- Analysis of Myristoylated and Palmitoylated Src Family Proteins -- Analysis of Myristoylated and Palmitoylated Src Family Proteins -- Ultracentrifugation Technique for Measuring the Binding of Peptides and Proteins to Sucrose-Loaded Phospholipid Vesicles -- Ultracentrifugation Technique for Measuring the Binding of Peptides and Proteins to Sucrose-Loaded Phospholipid Vesicles -- Biochemical and Biological Analyses of Farnesyl-Protein Transferase Inhibitors -- Biochemical and Biological Analyses of Farnesyl-Protein Transferase Inhibitors -- Identification and Characterization of Small GTPase-Associated Kinases -- Identification and Characterization of Small GTPase-Associated Kinases -- Functional Studies of Dual-Specificity Phosphatases -- Functional Studies of Dual-Specificity Phosphatases.

Transmembrane Signaling Protocols brings together a collection of practical, cutting-edge techniques for the study of cell signaling. Written by acknowledged leaders in the field, the protocols provide detailed, step-by-step instructions, helpful notes, and troubleshooting tips that make even the most powerful of the latest techniques readily reproducible. The protocols presented include the use of peptide libraries to study transmembrane signaling; the use of single-cell assays to analyze signal transduction pathways; the reconstitution of signaling complexes; methods for analyzing protein-protein interactions, and more. Introductory reviews explain the basic theory and enable researchers new to the area to rapidly gain understanding, as well as command of the practical knowledge and expertise afforded by the protocols. The time-tested techniques collected in Transmembrane Signaling Protocols make available to all researchers the many state-of-the-art techniques that have recently led to landmark discoveries in transmembrane signaling. The book is destined to become a standard resource for both conceptual information and the practical detail needed to carry out and interpret laboratory procedures in this dramatically advancing field.