In situ detection of S-glutathionylated proteins following glutaredoxin-1 catalyzed cysteine derivatization

Biochimica Et Biophysica Acta
Niki L ReynaertYvonne Janssen-Heininger

Abstract

S-glutathionylation is rapidly emerging as an important post-translational modification, responsible for transducing oxidant signals. However, few approaches are available that allow visualization of glutathione mixed disulfides in intact cells. We describe here a glutaredoxin1-dependent cysteine derivatization and labeling approach, in order to visualize S-glutathionylation patterns in situ. Using this new method, marked S-glutathionylation was observed in epithelial cells, which was predominant at membrane ruffles. As expected, the labeling intensity was further enhanced in response to bolus oxidant treatments, or in cells overexpressing Nox1 plus its coactivators. In addition, manipulation of endogenous levels of glutaredoxin-1 via RNAi, or overexpression resulted in altered sensitivity to H2O2 induced formation of glutathione mixed disulfides. Overall, the derivatization approach described here preferentially detects S-glutathionylation and provides an important means to visualize this post-translational modification in sub-cellular compartments and to investigate its relation to normal physiology as well as pathology.

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Citations

Apr 17, 2009·Biochemistry·Caroline Kumsta, Ursula Jakob
Aug 19, 2006·Proceedings of the National Academy of Sciences of the United States of America·Niki L ReynaertYvonne M W Janssen-Heininger
Jul 19, 2013·The Journal of Biological Chemistry·Young-Mi Go, Dean P Jones
Jul 29, 2009·Antioxidants & Redox Signaling·Suzy A A Comhair, Serpil C Erzurum
Jan 18, 2011·Antioxidants & Redox Signaling·Ying XiongDanyelle M Townsend
Nov 17, 2011·Antioxidants & Redox Signaling·Esther JortzikKatja Becker
Oct 18, 2012·Circulation. Cardiovascular Genetics·Christopher I Murray, Jennifer E Van Eyk
Sep 19, 2006·American Journal of Respiratory Cell and Molecular Biology·Niki L ReynaertYvonne M W Janssen-Heininger
Jun 12, 2010·American Journal of Respiratory Cell and Molecular Biology·Scott W AesifYvonne M W Janssen-Heininger
Apr 2, 2011·American Journal of Respiratory Cell and Molecular Biology·Ine KuipersNiki L Reynaert
Jun 23, 2012·PloS One·Ine KuipersNiki L Reynaert
Jul 28, 2013·International Journal of Molecular Sciences·Sarah UllevigReto Asmis
Jul 19, 2011·Free Radical Biology & Medicine·Scott W AesifYvonne M W Janssen-Heininger
Jun 27, 2009·The American Journal of Pathology·Scott W AesifYvonne M W Janssen-Heininger
Sep 13, 2008·Molecular Aspects of Medicine·Nazzareno BallatoriChristine L Hammond
Apr 22, 2008·Free Radical Biology & Medicine·Yvonne M W Janssen-HeiningerAlbert van der Vliet
Aug 19, 2007·Free Radical Biology & Medicine·Isabella Dalle-DonneAldo Milzani
Apr 5, 2013·Journal of Cellular Biochemistry·Yvonne M W Janssen-HeiningerVikas Anathy
Jan 30, 2013·Critical Reviews in Biochemistry and Molecular Biology·Young-Mi Go, Dean P Jones
Feb 19, 2013·Biochimica Et Biophysica Acta·Pietro Ghezzi
Dec 21, 2007·Antioxidants & Redox Signaling·Isabella Dalle-DonneRanieri Rossi
May 21, 2011·Antioxidants & Redox Signaling·Sebastian KehrKatja Becker
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Jun 15, 2021·Antioxidants & Redox Signaling·Moua Yang, Robert Flaumenhaft
Jan 23, 2018·Journal of Proteome Research·Fan YangHaiteng Deng

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