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.
© 2015 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Acetates / chemistry
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Benzothiazoles / chemistry
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Biotin / chemistry
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Cysteine / chemistry
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Cysteine / metabolism
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Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / analysis*
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Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / chemistry
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Human Umbilical Vein Endothelial Cells / cytology
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Human Umbilical Vein Endothelial Cells / drug effects
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Human Umbilical Vein Endothelial Cells / metabolism*
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Humans
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Hydrogen Sulfide / chemistry
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Hydrogen Sulfide / pharmacology*
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Jurkat Cells
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Microscopy, Fluorescence
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Oxidation-Reduction
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Protein Processing, Post-Translational*
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Serum Albumin, Bovine / analysis*
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Serum Albumin, Bovine / chemistry
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Signal Transduction
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Spectrometry, Mass, Electrospray Ionization
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Staining and Labeling / methods
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Streptavidin / chemistry
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Sulfides / chemistry
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Sulfides / metabolism
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Sulfides / pharmacology*
Substances
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Acetates
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Benzothiazoles
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Sulfides
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methylsulfonyl benzothiazole
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persulfides
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Serum Albumin, Bovine
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Biotin
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Streptavidin
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Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)
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Cysteine
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cyanoacetic acid
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sodium sulfide
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Hydrogen Sulfide