An outstanding collection of both classic and cutting-edge techniques for the detection and measurement of mitogen-activated protein kinase (MAPK) signaling cascades. The protocols includes methods for the determination of the subcellular localization of these components, the structural and biophysical analysis of the components, and identification of novel components of known and unknown signaling cascades. Additional methods examine the upstream mechanisms of activation of MAPK cascades by various receptors, the mechanisms involved in the down regulation of the MAPK cascades, and identification of targets of the MAPK cascades. A number of techniques use inhibitors in research on MAPK and transgenic mice for studies of MAPK signaling. The techniques offer biochemists, cell biologists, physicians, and biotechnologists all the essential laboratory techniques needed in to conduct productive studies of MAPK signaling in health and disease, in measuring the influence of drugs, and in a broad range of experimental systems.

Table of Contents:
The ERK Cascade As a Prototype of MAPK Signaling Pathways Hadara Rubinfeld and Rony Seger Determination of ERK Activity: Antiphospho-ERK Antibodies, In Vitro Phosphorylation, and In-Gel Kinase Assay Sarah Kraus and Rony Seger Detection of ERK1/2 Activities Using Affinity Reagents Rafael Pulido, ngel Zuniga, and Axel Ullrich Activation of SAPKs/JNKs and p38s In Vitro John M. Kyriakis, Hong Liu, and Deborah N. Chadee Investigating the Cellular BMK1/ERK5 Signaling Pathway Richard I. Tapping, Kato Yutaka, Ta-Hsiang Chao, Masaaki Hayashi, Jeng-Fan Lo, Sung-Woo Kim, and Jiing-Dwan Lee Pull-Down Assays for Guanosine 5'-Triphosphate-Bound Ras-Like Guanosine 5'-Triphosphatases Miranda van Triest and Johannes L. Bos Regulation of MAPK Cascades by Protein Tyrosine Phosphatases Josema Torres, Carmen Blanco-Aparicio, and Rafael Pulido Use of Inhibitors in the Study of MAPK Signaling Yoav D. Shaul and Rony Seger Structure of MAPKs Elizabeth J. Goldsmith, Melanie H. Cobb, and Chung-I Chang Subcellular Localization of MAPKs Makoto Adachi and Eisuke Nishida Study of MAPK Signaling Using Knockout Mice Gilles Pages and Jacques Pouyssegur Computer Simulation of MAPK Signal Transduction Baltazar D. Aguda and Herbert M. Sauro Signaling by Growth Factor Receptors Bose S. Kochupurakkal and Yosef Yarden Activation of MAPK by G Protein-Coupled Receptors Piero Crespo and J. Silvio Gutkind Identification of MAPK Substrates by Expression Screening With Solid-Phase Phosphorylation Rikiro Fukunaga and Tony Hunter Study of Substrate Specificity of MAPKs Using Oriented Peptide Libraries Michael B. Yaffe Ras Signaling Pathway for Analysis of Protein-Protein Interactions in Yeast and Mammalian Cells Ami Aronheim Methods in Functional Proteomics: Two-Dimensional Polyacrylamide Gel Electrophoresis With Immobilized pH Gradients, In-Gel Digestion, and Identification of Proteins by Mass Spectrometry Karine R. Bernard, Karen R. Jonscher, Katheryn A. Resing, and Natalie G. Ahn Practical Methods for Deuterium Exchange/Mass Spectrometry Andrew N. Hoofnagle, Katheryn A. Resing, and Natalie G. Ahn Use of Xenopus Oocytes and Early Embryos to Study MAPK Signaling Eusebio Perdiguero and Angel R. Nebreda MAPK Cascades in the Brain: Lessons From Learning Diego E. Berman and Yadin Dudai Index