Analysis of the redox regulation of protein tyrosine phosphatase superfamily members utilizing a cysteinyl-labeling assay

Boivin, B., Tonks, N. K. (2010) Analysis of the redox regulation of protein tyrosine phosphatase superfamily members utilizing a cysteinyl-labeling assay. In: Thiol Redox Transitions in Cell Signaling, Part B: Cellular Localization and Signaling. Methods in Enzymology, 474 . Elsevier Academic Press Inc, San Diego, pp. 35-50. ISBN 0076-6879978-0-12-381003-8

URL: http://www.ncbi.nlm.nih.gov/pubmed/20609903
DOI: 10.1016/s0076-6879(10)74003-9

Abstract

The catalytic activity of protein tyrosine phosphatase (PIP) superfamily members is regulated by the reversible oxidation of their invariant catalytic Cys residue in vivo. Transient and specific regulation of PIP activity by reactive oxygen species (ROS) attenuates dephosphorylation and, thereby, promotes phosphorylation, hence facilitating signal transduction. We have recently developed a modified cysteinyl-labeling assay [Boivin, B., Zhang, S., Arbiser, J. L., Zhang, Z. Y., and Tonks, N. K. (2008). Proc. Natl. Acad. Sci. USA 105, 9959-9964] that showed broad selectivity in detecting reversible oxidation of members from different PIP subclasses in platelet-derived growth factor (PDGF)-BB overexpressing cells. Herein, we applied this assay, which utilizes the unique chemistry of the invariant catalytic Cys residue to enrich and identify PTPs that are reversibly oxidized upon acute growth factor stimulation. Performing the cysteinyl-labeling assay with Rat-1 fibroblasts enabled us to capture both PTEN and SHP-2 as a consequence to acute PDGF-BB stimulation. Given the ability of this assay to detect reversible oxidation of a broad array of members of the PIP family, we anticipate that it should permit profiling of the entire ROS-regulated PTPome in a wide array of signaling paradigms.

Item Type: Book Section
Uncontrolled Keywords: REVERSIBLE OXIDATION HYDROGEN-PEROXIDE SULFENIC ACID GROWTH-FACTOR IN-VIVO SIGNAL-TRANSDUCTION INTACT-CELLS PTP-ALPHA INACTIVATION 1B
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
diseases & disorders > pulmonary disease > oxidative stress
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > protein tyrosine phosphatase
CSHL Authors:
Communities: CSHL labs > Tonks lab
Depositing User: Matt Covey
Date: 2010
Date Deposited: 18 Dec 2013 19:54
Last Modified: 18 Dec 2013 19:54
Related URLs:
URI: http://repository.cshl.edu/id/eprint/29009

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving