Protein Phosphatase Protocols

1. Front Matter

   Pages i-xv

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2. Analysis of Protein Phosphatases

   • Shirish Shenolikar

   Pages 1-8

3. A Brief Introduction to the Protein Phosphatase Families

   • Tomas Mustelin

   Pages 9-22

4. Small-Molecule Inhibitors of Ser/Thr Protein Phosphatases

   • Mark Swingle, Li Ni, Richard E. Honkanen

   Pages 23-38

5. Synthesis and Use of the Protein Phosphatase Affinity Matrices Microcystin-Sepharose and Microcystin-Biotin-Sepharose

   • Greg B. G. Moorhead, Timothy A. J. Haystead, Carol MacKintosh

   Pages 39-45

6. Utilizing Protein Phosphatase Inhibitors to Define PP2A as a Regulator of Ataxia-Telangiectasia Mutated

   • Aaron A. Goodarzi, Pauline Douglas, Greg B. G. Moorhead, Susan P. Lees-Miller

   Pages 47-59

7. An Automated Fluorescence-Based Method for Continuous Assay of PP2A Activity

   • Adam M. Wegner, Jamie L. McConnell, Randy D. Blakely, Brian E. Wadzinski

   Pages 61-69

8. An In Vivo Assay to Quantify Stable Protein Phosphatase 2A (PP2A) Heterotrimeric Species

   • Matthew S. Gentry, Richard L. Hallberg, David C. Pallas

   Pages 71-83

9. Mutagenesis and Expression of the Scaffolding Aα and Aβ Subunits of PP2A

   • Ralf Ruediger, Jin Zhou, Gernot Walter

   Pages 85-99

10. Isolation and Characterization of PP2A Holoenzymes Containing FLAG-Tagged B Subunits

    • Deanna G. Adams, Brian E. Wadzinski

    Pages 101-111

11. Purification of PP2A Holoenzymes by Sequential Immunoprecipitation with Anti-Peptide Antibodies

    • Gernot Walter, Jin Zhou, Ralf Ruediger

    Pages 113-126

12. Purification of PP2Ac from Bovine Heart

    • Hue T. Tran, Tony S. Ferrar, Anne Ulke-Lemée, Greg B. G. Moorhead

    Pages 127-132

13. Visualization of Intracellular PP1 Targeting Through Transiently and Stably Expressed Fluorescent Protein Fusions

    • Laura Trinkle-Mulcahy, Janet Chusainow, Yun Wah Lam, Sam Swift, Angus Lamond

    Pages 133-154

14. Yeast Two-Hybrid Screens to Identify Drosophila PP1-Binding Proteins

    • Daimark Bennett, Luke Alphey

    Pages 155-179

15. Identification of Cellular Protein Phosphatase-1 Regulators

    • David W. Roadcap, Matthew H. Brush, Shirish Shenolikar

    Pages 181-196

16. Assay for Three-Way Interaction of Protein Phosphatase-1 (Glc7) With Regulatory Subunits Plus Phosphatase Inhibitor-2

    • Masumi Eto, David L. Brautigan

    Pages 197-208

17. Phosphorylation of the Protein Phosphatase Type 1 Inhibitor Protein CPI-17 by Protein Kinase C

    • Michael P. Walsh, Marija Susnjar, Jingti Deng, Cindy Sutherland, Enikő Kiss, David P. Wilson

    Pages 209-223

18. Purification of Smooth Muscle Myosin Phosphatase Using a Thiophosphorylated Myosin Light-Chain-Affinity Resin

    • Meredith Borman, Justin MacDonald

    Pages 225-233

19. Proteins Interacting With Saccharomyces cerevisiae Type 1 Protein Phosphatase Catalytic Subunit Identified by Single-Step Affinity Purification and Mass Spectrometry

    • Edmund P. Walsh, Douglas J. Lamont, Kenneth A. Beattie, Michael J. R. Stark

    Pages 235-246

20. Expression of Protein Histidine Phosphatase in Escherichia coli, Purification, and Determination of Enzyme Activity

    • Nicole Bäumer, Anette Mäurer, Josef Krieglstein, Susanne Klumpp

    Pages 247-260

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.

• Cell signaling networks
• Drosophilia
• Drug targets
• Phosphatase families
• Proteomics
• Termination
• Vivo
• Yeast
• biochemistry
• bioinformatics
• escherichia coli
• protein sequence
• saccharomyces cerevisiae

• Department of Biological Sciences, University of Calgary, Calgary, Canada

  Greg Moorhead

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