Protein Phosphatase Protocols PDF Book 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.
Table of Contents
Preface ………………………………………………………………………………………………..v
Contributors ………………………………………………………………………………………..xi
1 Analysis of Protein Phosphatases:
Toolbox For Unraveling Cell Signaling Networks
2 A Brief Introduction to the Protein Phosphatase Families
3 Small-Molecule Inhibitors of Ser/Thr Protein Phosphatases:
Specificity, Use, and Common Forms of Abuse
4 Synthesis and Use of the Protein Phosphatase Affinity Matrices
Microcystin–Sepharose and Microcystin–Biotin–Sepharose
5 Utilizing Protein Phosphatase Inhibitors to Define PP2A
as a Regulator of Ataxia-Telangiectasia Mutated (ATM)
6 An Automated Fluorescence-Based Method
for Continuous Assay of PP2A Activity
7 An In Vivo Assay to Quantify Stable Protein
Phosphatase 2A (PP2A) Heterotrimeric Species
8 Mutagenesis and Expression of the Scaffolding Aα and Aβ
Subunits of PP2A: Assays for Measuring Defects
in Binding of Cancer-Related Aα and Aβ Mutants
to the Regulatory B and Catalytic C Subunits
9 Isolation and Characterization of PP2A Holoenzymes Containing
FLAG-Tagged B Subunits
10 Purification of PP2A Holoenzymes by Sequential
Immunoprecipitation with Anti-Peptide Antibodies
11 Purification of PP2Ac from Bovine Heart
12 Visualization of Intracellular PP1 Targeting Through Transiently
and Stably Expressed Fluorescent Protein Fusions
13 Yeast Two-Hybrid Screens To Identify
Drosophila PP1-Binding Proteins
14 Identification of Cellular Protein Phosphatase-1 Regulators
15 Assay for Three-Way Interaction of Protein Phosphatase-1 (Glc7)
with Regulatory Subunits Plus Phosphatase Inhibitor-2
16 Phosphorylation of the Protein Phosphatase Type 1
Inhibitor Protein CPI-17 by Protein Kinase C
17 Purification of Smooth Muscle Myosin Phosphatase Using a
Thiophosphorylated Myosin Light-Chain-Affinity Resin
18 Proteins Interacting with Saccharomyces cerevisiae Type 1
Protein Phosphatase Catalytic Subunit Identified
by Single-Step Affinity Purification and Mass Spectrometry
19 Expression of Protein Histidine Phosphatase in Escherichia coli,
Purification, and Determination of Enzyme Activity
20 The Use of RNA Interference to Analyze Protein
Phosphatase Function In Mammalian Cells
21 Recognition of a PP2C Interaction Motif
in Several Plant Protein Kinases
22 Use of Yeast Genetic Tools to Define Biological Roles
of Novel Protein Phosphatases
23 Targeting of PP2C in Budding Yeast
24 Phosphatase Targets in TOR Signaling
25 Functional Characterization of the Small CTD Phosphatases
26 Genome-Scale Discovery and Characterization
of Class-Specific Protein Sequences: An Example
Using the Protein Phosphatases of Arabidopsis thaliana
27 Yeast Substrate-Trapping System for Isolating Substrates
of Protein Tyrosine Phosphatases