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

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / chemistry
  • Benzothiazoles / chemistry
  • Biotin / chemistry
  • Cysteine / chemistry
  • Cysteine / metabolism
  • Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / analysis*
  • Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / chemistry
  • Human Umbilical Vein Endothelial Cells / cytology
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Human Umbilical Vein Endothelial Cells / metabolism*
  • Humans
  • Hydrogen Sulfide / chemistry
  • Hydrogen Sulfide / pharmacology*
  • Jurkat Cells
  • Microscopy, Fluorescence
  • Oxidation-Reduction
  • Protein Processing, Post-Translational*
  • Serum Albumin, Bovine / analysis*
  • Serum Albumin, Bovine / chemistry
  • Signal Transduction
  • Spectrometry, Mass, Electrospray Ionization
  • Staining and Labeling / methods
  • Streptavidin / chemistry
  • Sulfides / chemistry
  • Sulfides / metabolism
  • Sulfides / pharmacology*

Substances

  • Acetates
  • Benzothiazoles
  • Sulfides
  • methylsulfonyl benzothiazole
  • persulfides
  • Serum Albumin, Bovine
  • Biotin
  • Streptavidin
  • Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)
  • Cysteine
  • cyanoacetic acid
  • sodium sulfide
  • Hydrogen Sulfide