Protein Phosphatase Protocols

ISBN-13: 9781617377334 / Angielski / Miękka / 2010 / 387 str.

Greg Moorhead

Opis

Protein Phosphatase Protocols presents a broad range of protocols for the study of protein phosphatases, all written by experts and innovators from phosphatase laboratories around the world. This volume is a compendium of resources for the study of protein phosphatases and their potential as drug targets. Experimental methodologies are taken from proteomics, bioinformatics, genomics, biochemistry, RNAi, and genetics. Included protocols utilize several model organisms, including yeast, Drosophila, and various plant and human cells. Protein Phosphatase Protocols provides a concise introduction to protein phosphatase research, with particular attention to phosphatase families. Assays for identification, quantification, purification, and functional characterization are presented throughout in specific detail.

Spis treści

Contents Preface………………………………………………………………………………v Contributors…………………………………………………………………………xi Analysis Of Protein Phosphatases: Toolbox For Unraveling Cell Signaling Networks. Shirish Shenolikar………………………………………………………..……X A Brief Introduction To The Protein Phosphatase Families. Tomas Mustelin…………………………………………………………..……X Small Molecule Inhibitors of Ser/thr Protein Phosphatases: Specificity, Use and Common Forms of Abuse. Mark Swingle, Li Ni and Richard E. Honkanen…………………………….X Synthesis and Use of the Protein Phosphatase Affinity Matrices Microcystin- and Microcystin-biotin-Sepharose Greg Moorhead, Timothy AJ Haystead and Carol MacKintosh………..…X Utilizing Protein Phosphatase Inhibitors To Define PP2A As A Regulator Of Ataxia-Telangiectasia Mutated (ATM). Aaron A. Goodarzi, Pauline Douglas, Greg B.G. Moorhead and Susan P. Lees-Miller……………………………………………………………………….X An Automated, Fluorescence-Based Method For Continuous Assay Of PP2A Activity. Adam M. Wegner, Jamie L. McConnell, Randy D. Blakely, and Brian E. Wadzinski…………….…………………….…………………………...……….X An In Vivo Assay To Quantify Stable Protein Phosphatase 2A (PP2A) Heterotrimeric Species. Matthew S. Gentry, Richard L. Hallberg, and David C. Pallas.…………….X Mutagenesis And Expression Of The Scaffolding AaAnd Ab Subunits Of PP2A; Assays For Measuring Defects In Binding Of Cancer-Related Aa And Ab Mutants To The Regulatory B And Catalytic C Subunits. Ralf Ruediger, Jin Zhou, and Gernot Walter……………….…………..…….X Isolation and Characterization of PP2A Holoenzymes Containing FLAG-tagged B Subunits. Deanna G. Adams and Brian E. Wadzinski………………….………..…..….X Purification Of PP2A Holoenzymes By Sequential Immunoprecipitation With Anti-Peptide Antibodies. Gernot Walter, Jin Zhou, and Ralf Ruediger………………………………....X Purification Of PP2Ac From Bovine Heart. Hue T Tran, Tony S Ferrar, Anne Ulke-Lemée and Greg BG Moorhead……………………chr(133David W. Roadcap, Matthew H. Brush, and Shirish Shenolikar……….…X Assay For 3-Way Interaction Of Protein Phosphatase-1 (Glc7) With Regulatory Subunits Plus Phosphatase Inhibitor-2. Masumi Eto and David L. Brautigan………………………………….………X Phosphorylation Of The Protein Phosphatase Type 1 Inhibitor Protein, CPI-17, By Protein Kinase C. Michael P. Walsh, Marija Susnjar, Jingti Deng, Cindy Sutherland, Enikó Kiss, and David P. Wilson………………………………………………………X Purification Of Smooth Muscle Myosin Phosphatase Using A Thiophosphorylated Myosin Light Chain-Affinity Resin. Merideth Borman and Justin MacDonald……………………………….…….X Proteins Interacting With Saccharomyces Cerevisiae Type 1 Protein Phosphatase Catalytic Subunit Identified By Single-Step Affinity Purification And Mass Spectrometry.