S-Glutathionylation Enhances Human Cystathionine β-Synthase Activity Under Oxidative Stress Conditions

Authors

Wei-Ning Niu, The Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi’an, China
Pramod Kumar Yadav, Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan
Jiri Adamec, University of Nebraska-LincolnFollow
Ruma Banerjee, Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan

Document Type

Article

Date of this Version

2015

Citation

Antioxid. Redox Signal. 22, 350–361.

Comments

U.S. Government Work

Abstract

Aims: Cystathionine β-synthase (CBS) catalyzes the first and rate-limiting step in the two-step trans-sulfuration pathway that converts homocysteine to cysteine. It is also one of three major enzymes responsible for the biogenesis of H₂S, a signaling molecule. We have previously demonstrated that CBS is activated in cells challenged by oxidative stress, but the underlying molecular mechanism of this regulation has remained unclear. Results: Here, we demonstrate that S-glutathionylation of CBS enhances its activity ~2-fold in vitro. Loss of this post-translational modification in the presence of dithiothreitol results in reversal to basal activity.

Cys346 was identified as the site for S-glutathionylation by a combination of mass spectrometric, mutagenesis, and activity analyses. To test the physiological relevance of S-glutathionylation-dependent regulation of CBS, HEK293 cells were oxidatively challenged with peroxide, which is known to enhance the trans-sulfuration flux. Under these conditions, CBS glutathionylation levels increased and were correlated with a ~3-fold increase in CBS activity. Innovation: Collectively, our results reveal a novel post-translational modification of CBS, that is, glutathionylation, which functions as an allosteric activator under oxidative stress conditions permitting enhanced synthesis of both cysteine and H₂S. Conclusions: Our study elucidates a molecular mechanism for increased cysteine and therefore glutathione, synthesis via glutathionylation of CBS. They also demonstrate the potential for increased H₂S production under oxidative stress conditions, particularly in tissues where CBS is a major source of H₂S.