Beschreibung
A major mechanism by which cells regulate protein function is to place phosphate groups on serine and threonine residues. Though the steady-state level of protein phosphorylation depends on the relative activities of both kinases and phosphatases, a much greater effort has previously gone into the study of the former that the latter. Today, however, there is an increasing appreciation for the role that protein phosphatases play in the dynamic p- cess of protein phosphorylation. To date, there are four major types of protein serine/threonine phosphatase catalytic subunits, designated protein phosphatase type 1, 2A, 2B, and 2C. Each has been identified by the techniques of protein chemistry and enzymology and can be distinguished from one another by their preference for specific substrates as well as their sensitivity to certain acti- tors and inhibitors. Protein Phosphatase Protocols has been assembled in response to the growing interest these enzymes are receiving. The goal of this compilation is to provide a "how-to" experimental guide to aid newcomers as well as s- soned veterans in their research endeavors, thus further contributing towards our ever increasing knowledge of serine/threonine phosphatases. What you have before you contains contributions by many of the current and emerging leaders in the field. To highlight just a few, these chapters c- tain step-by-step information on how to isolate novel phosphatases and re- latory subunits, assay for activity, and generate immunological reagents for both biochemical and biological characterization of these enzymes.
Autorenporträt
InhaltsangabeProkaryotic Protein-Serine/Threonine Phosphatases P. J. Kennelly Protein Phosphatase Type 1 and 2A Assays S. D. Killilea, Q.Cheng, and Z. X. Wang. Analyzing Gene Expression with the Use of Serine/Threonine Phosphatase Inhibitors A. H. Shönthal Inhibitor1, a Regulator of Protein Phosphatase1 Function J. H. Connor, H. Q., C. Oliver, and S. Shenolikar I1PP2A and I2PP2A: Two Potent Protein Phosphatase 2A-Specific Inhibitor Proteins M. L. and Z. Damuni Control of PP1 Activity Through Phosphorylation by Cyclin-Dependent Kinases N. Berndt. Regulation of Neuronal PP1 and PP2A During Development E. Collins and A.T. R. Sim PTPA Regulating PP2A as a Dual Specificity Phosphatase V.Janssens, C. Van Hoof, W. Merlevede, and J. Goris Microinjection and Immunological Methods in the Analysis of Type 1 and Type 2A Protein Phosphatases from Mammalian Cells P.Turowski and N. J. C. Lamb Use of Immunocomplexed Substrate for Detecting PP1 Activity J. W. Ludlow, D. A. Nelson, and N. A. Krucher The Biochemical Identification and Characterization of New Species of Protein Phosphatase 1 M. Beullens, W. Stalmans, and M. Bollen The Relationship between insulin signaling and protein phosphatase 1 activation L. Ragolia and N. Begum Analysis of the isoforms of protein phosphatase (PP1) isoforms with polyclonal peptide antibodies M. Tognarini and E. Villa-Moruzzi Expression of mouse protein phosphatase 2C in Eschericia coli and COS 7 cells Takayasu Kobayashi, Kazuyuki Kusuda, Motoko Ohnishi, Naoki Chida, and Shinri Tamura Expression of Functional protein phosphatase 1 catalytic subunit in Eschericia coli M. Dohadwala and N. Berndt Protein Phosphatase 2A and Protein Phosphatase X Genes in Arabidopsis thaliana G. Pujol, A. Ferrer, and J.Ariño Separation of Protein Phosphatase Type 2C Isozymes by Chromatography on Blue Sepharose S. Klumpp and D. Selke Chromatographic Isolation of PP2A from Limulus Lateral Eyes: Conventional and Small Scale Methods S. C. Edwards, T. B. Van Dyke, T. H. Van Dyke, and D. L. Brautigan Purification and Assay of the Ptc/Tpd1 Protein Phosphatase 2C from the yeast Saccharomyces cerevisiae M. K. Robinson and E. M. Phizicky Molecular cloning of protein phosphatase type 2C isoforms from retinal cDNA Da. Selke, S. Klumpp, B. Kaupp, and A. Baumann Analysis of Protein Interactions Between Protein Phosphatase 1 and Noncatalytic Subunits using the Yeast Two-Hybrid Assay N. T. Ramaswamy, B. K. Dalley, and J. F. Cannon Identifying Protein Phosphatase 2A Interacting Proteins Using the Yeast Two-hybrid method B.McCright and D.M. Virshup Protein phosphatase 2A regulatory subunits: cDNA cloning and analysis of mRNA expression J. A. Zaucha, R. S. Westphal, and B. E. Wadzinski Synthetic Lethal Screening in Protein Phosphatase Pathways J. Zheng and J. F. Cannon The Search for the Biological Function of Novel Yeast ser/thr Phosphatases J. Ariño, F. Posas, and J. Clotet Index
