Use of the "tag-switch" method for the detection of protein S-sulfhydration.
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.
Department of Chemistry, Washington State University, Pullman, Washington, USA.
Chung-Min
CM
Park
Department of Chemistry and Pharmacy, University of Erlangen-Nuremberg, Erlangen, Germany.
Igor
I
Macinkovic
Department of Chemistry and Pharmacy, University of Erlangen-Nuremberg, Erlangen, Germany. Electronic address: milos.filipovic@fau.de.
Milos R
MR
Filipovic
Department of Chemistry, Washington State University, Pullman, Washington, USA. Electronic address: mxian@wsu.edu.
Ming
M
Xian
D000085:Acetates / Q000737:chemistry; D052160:Benzothiazoles / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000032:analysis* / Q000737:chemistry; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006862:Hydrogen Sulfide / Q000737:chemistry / Q000494:pharmacology*; D019169:Jurkat Cells; D008856:Microscopy, Fluorescence; D010084:Oxidation-Reduction; D011499:Protein Processing, Post-Translational; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000737:chemistry; D015398:Signal Transduction; D021241:Spectrometry, Mass, Electrospray Ionization; D013194:Staining and Labeling / Q000379:methods; D019809:Streptavidin / Q000737:chemistry; D013440:Sulfides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*
D000085:Acetates; D052160:Benzothiazoles; D013440:Sulfides; C576094:methylsulfonyl benzothiazole; C051552:persulfides; D012710:Serum Albumin, Bovine; D001710:Biotin; D019809:Streptavidin; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); D003545:Cysteine; C007170:cyanoacetic acid; C033479:sodium sulfide; D006862:Hydrogen Sulfide
Hydrogen sulfide; Persulfide; Posttranslational modification; Protein; Redox signaling; S-Sulfhydration