Persulfide reactivity in the detection of protein s-sulfhydration

ACS Chem Biol. 2013;8(6):1110-6. doi: 10.1021/cb4001052. Epub 2013 Apr 12.


Hydrogen sulfide (H2S) has emerged as a new member of the gaseous transmitter family of signaling molecules and appears to play a regulatory role in the cardiovascular and nervous systems. Recent studies suggest that protein cysteine S-sulfhydration may function as a mechanism for transforming the H2S signal into a biological response. However, selective detection of S-sulfhydryl modifications is challenging since the persulfide group (RSSH) exhibits reactivity akin to other sulfur species, especially thiols. A modification of the biotin switch technique, using S-methyl methanethiosulfonate (MMTS) as an alkylating reagent, was recently used to identify a large number of proteins that may undergo S-sulfhydration, but the underlying mechanism of chemical detection was not fully explored. To address this key issue, we have developed a protein persulfide model and analogue of MMTS, S-4-bromobenzyl methanethiosulfonate (BBMTS). Using these new reagents, we investigated the chemistry in the modified biotin switch method and examined the reactivity of protein persulfides toward different electrophile/nucleophile species. Together, our data affirm the nucleophilic properties of the persulfide sulfane sulfur and afford new insights into protein S-sulfhydryl chemistry, which may be exploited in future detection strategies.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alkylating Agents / chemistry
  • Biochemistry / methods
  • Biotin / chemistry
  • Hydrogen Sulfide / chemistry
  • Mesylates / chemistry
  • Methyl Methanesulfonate / analogs & derivatives
  • Methyl Methanesulfonate / chemistry
  • Protein S / chemistry*
  • Sulfhydryl Compounds / analysis
  • Sulfides / analysis*


  • Alkylating Agents
  • Mesylates
  • Protein S
  • Sulfhydryl Compounds
  • Sulfides
  • persulfides
  • methyl methanethiosulfonate
  • methanethiosulfonate
  • Biotin
  • Methyl Methanesulfonate
  • Hydrogen Sulfide