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

N.L. Reynaert, K. Ckless, A.S. Guala, E.F. Wouters, A. van der Vliet, Y.M. Janssen-Heininger*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review


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.
Original languageEnglish
Pages (from-to)380-387
JournalBiochimica et Biophysica Acta-general Subjects
Issue number3
Publication statusPublished - 1 Jan 2006

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