Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Subscribe

Sci. STKE, 18 December 2007
Vol. 2007, Issue 417, p. pl8
[DOI: 10.1126/stke.4172007pl8]

PROTOCOLS

Identification of Redox-Active Cell-Surface Proteins by Mechanism-Based Kinetic Trapping

Ulla Schwertassek{dagger}, Lars Weingarten, and Tobias P. Dick*

Redox Regulation Research Group (A160), German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
{dagger}Present address: Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.

Abstract: A number of thiol-dependent oxidoreductases are released from cells and act on the cell surface. Correspondingly, several cell-surface processes appear to depend on catalyzed thiol-disulfide exchange, including integrin activation and the fusion of viral particles with the host membrane. Tumor cells frequently increase the abundance of secreted and cell-surface forms of particular oxidoreductases, and evidence suggests that oxidoreductases released from tumor cells promote growth and contribute to the remodeling of the cellular microenvironment. Few cell-surface or membrane proteins that are targeted by extracellular redox enzymes have been identified. One major reason for this slow progress is the highly transient nature of thiol-disulfide exchange, making its detection by conventional techniques difficult or impossible. Here we describe the application of an activity-based proteomics approach, also known as "mechanism-based kinetic trapping," to identify individual cell-surface target proteins that engage in disulfide exchange with thiol-dependent oxidoreductases. Although we have applied this approach to thioredoxin-1, it should also be applicable to other members of the thioredoxin superfamily whose activity is based on the CXXC active-site motif.

*Corresponding author. E-mail, t.dick{at}dkfz.de

Citation: U. Schwertassek, L. Weingarten, T. P. Dick, Identification of Redox-Active Cell-Surface Proteins by Mechanism-Based Kinetic Trapping. Sci. STKE 2007, pl8 (2007).

Read the Full Text

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Reduced Monomeric CD4 Is the Preferred Receptor for HIV.
L. J. Matthias, I. Azimi, C. A. Tabrett, and P. J. Hogg (2010)
J. Biol. Chem. 285, 40793-40799
   Abstract »    Full Text »    PDF »
Disulfide Bond That Constrains the HIV-1 gp120 V3 Domain Is Cleaved by Thioredoxin.
I. Azimi, L. J. Matthias, R. J. Center, J. W. H. Wong, and P. J. Hogg (2010)
J. Biol. Chem. 285, 40072-40080
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882