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Methods Enzymol. 2015;555:39-56. doi: 10.1016/bs.mie.2014.11.033. Epub 2015 Jan 8.

Use of the "tag-switch" method for the detection of protein S-sulfhydration.

Author information

1
Department of Chemistry, Washington State University, Pullman, Washington, USA.
2
Department of Chemistry and Pharmacy, University of Erlangen-Nuremberg, Erlangen, Germany.
3
Department of Chemistry and Pharmacy, University of Erlangen-Nuremberg, Erlangen, Germany. Electronic address: milos.filipovic@fau.de.
4
Department of Chemistry, Washington State University, Pullman, Washington, USA. Electronic address: mxian@wsu.edu.

Abstract

Protein S-sulfhydration (i.e., converting protein cysteines -SH to persulfides -SSH) is a redox-based posttranslational modification. This reaction plays an important role in signaling pathways mediated by hydrogen sulfide or other reactive sulfane sulfur species. Recently, our laboratories developed a "tag-switch" method which can be used to selectively label and detect protein S-sulfhydrated residues. In this chapter, we provide a comprehensive summary of this method, including the design of the method, preparation of the reagents, validation on small-molecule substrates, as well as applications in protein labeling. Experimental protocols for the use of the method are described in details.

KEYWORDS:

Hydrogen sulfide; Persulfide; Posttranslational modification; Protein; Redox signaling; S-Sulfhydration

PMID:
25747474
DOI:
10.1016/bs.mie.2014.11.033
[Indexed for MEDLINE]

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