Unit

UNIT 11.7 Using Protein Kinase and Protein Phosphatase Inhibitors to Dissect Protein Phosphorylation Pathways

  1. James W. Karaszkiewicz1,
  2. Curtis J. Henrich2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im1107s22

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Karaszkiewicz, J. W. and Henrich, C. J. 2001. Using Protein Kinase and Protein Phosphatase Inhibitors to Dissect Protein Phosphorylation Pathways. Current Protocols in Immunology. 22:11.7:11.7.1–11.7.20.

Author Information

  1. 1

    Kirkegaard and Perry Laboratories, Gaithersburg, Maryland

  2. 2

    Life Technologies, Inc., Gaithersburg, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1997

This is not the most recent version of the article. View current version (1 APR 2011)

Abstract

It is often desirable to attempt to measure and to modulate the activity of the protein kinases and phosphatases responsible for a given phenotypic change. The protocols in this unit describe how to assay a variety of protein kinase activities in complex mixtures, such as whole cell lysates, in order to screen compounds as potential inhibitors of these activities. The effects of known or potential inhibitors can be evaluated by monitoring phenotypic changes in intact cells following treatment with the inhibitor. The activities of the major serine/threonine-specific protein phosphatases in most mammalian cells can be distinguished using a selective inhibitor, okadaic acid. Support protocols describe preparation of cell extracts needed for the kinase assays, and of 32P-labeled phosphorylase used in the inhibition assays.