Skip to main content
Article
Irreversible Thiol Oxidation in Carbonic Anhydrase III: Protection by S-Glutathiolation and Detection in Aging Rats
Biological Chemistry
  • Robert J. Mallis, Iowa State University
  • Michael J. Hamann, Iowa State University
  • Wei Zhao, Iowa State University
  • Tiequan Zhang, Iowa State University
  • Suzanne Hendrich, Iowa State University
  • James A. Thomas, Iowa State University
Document Type
Article
Publication Date
3-1-2002
DOI
10.1515/BC.2002.067
Abstract

Proteins with reactive sulfhydryls are central to many important metabolic reactions and also contribute to a variety of signal transduction systems. In this re- port, we examine the mechanisms of oxidative dam- age to the two reactive sulfhydryls of carbonic anhy- drase III. Hydrogen peroxide (H2O2), peroxy radicals, or hypochlorous acid (HOCl) produced irreversibly oxidized forms, primarily cysteine sulfinic acid or cys- teic acid, of carbonic anhydrase III if glutathione (GSH) was not present. When GSH was approximate- ly equimolar to protein thiols, irreversible oxidation was prevented. H2O2 and peroxyl radicals both gener- ated S-glutathiolated carbonic anhydrase III via par- tially oxidized protein sulfhydryl intermediates, while HOCl did not cause S-glutathiolation. Thus, oxidative damage from H2O2 or AAPH was prevented by protein S-glutathiolation, while a direct reaction between GSH and oxidant likely prevents HOCl-mediated pro- tein damage. In cultured rat hepatocytes, carbonic anhydrase III was rapidly S-glutathiolated by mena- dione. When hepatocyte glutathione was depleted, menadione instead caused irreversible oxidation. We hypothesized that normal depletion of glutathione in aged animals might also lead to an increase in irre- versible oxidation. Indeed, both total protein extracts and carbonic anhydrase III contained significantly more cysteine sulfinic acid in older rats compared to young animals. These experiments show that, in the absence of sufficient GSH, oxidation reactions lead to irreversible protein sulfhydryl damage in purified pro- teins, cellular systems, and whole animals.

Comments

This article is from Biological Chemistry, 383(3-4) 2002: 649-662. doi: 10.1515/BC.2002.067. Posted with permission. The final publication is available at www.degruyter.com.

Copyright Owner
Walter de Gruyter
Language
en
Date Available
2014-11-14
File Format
application/pdf
Citation Information
Robert J. Mallis, Michael J. Hamann, Wei Zhao, Tiequan Zhang, et al.. "Irreversible Thiol Oxidation in Carbonic Anhydrase III: Protection by S-Glutathiolation and Detection in Aging Rats" Biological Chemistry Vol. 383 Iss. 3-4 (2002) p. 649 - 662
Available at: http://works.bepress.com/suzanne_hendrich/17/