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

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.