Inactivation of thiol-dependent enzymes by hypothiocyanous acid: role of sulfenyl thiocyanate and sulfenic acid intermediates

Research output: Contribution to journalJournal articleResearchpeer-review

Tessa J Barrett, David I Pattison, Stephen E Leonard, Kate S Carroll, Michael Jonathan Davies, Clare Louise Hawkins

Myeloperoxidase (MPO) forms reactive oxidants including hypochlorous and hypothiocyanous acids (HOCl and HOSCN) under inflammatory conditions. HOCl causes extensive tissue damage and plays a role in the progression of many inflammatory-based diseases. Although HOSCN is a major MPO oxidant, particularly in smokers, who have elevated plasma thiocyanate, the role of this oxidant in disease is poorly characterized. HOSCN induces cellular damage by targeting thiols. However, the specific targets and mechanisms involved in this process are not well defined. We show that exposure of macrophages to HOSCN results in the inactivation of intracellular enzymes, including creatine kinase (CK) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In each case, the active-site thiol residue is particularly sensitive to oxidation, with evidence for reversible inactivation and the formation of sulfenyl thiocyanate and sulfenic acid intermediates, on treatment with HOSCN (less than fivefold molar excess). Experiments with DAz-2, a cell-permeable chemical trap for sulfenic acids, demonstrate that these intermediates are formed on many cellular proteins, including GAPDH and CK, in macrophages exposed to HOSCN. This is the first direct evidence for the formation of protein sulfenic acids in HOSCN-treated cells and highlights the potential of this oxidant to perturb redox signaling processes.

Original languageEnglish
JournalFree Radical Biology & Medicine
Volume52
Issue number6
Pages (from-to)1075-85
Number of pages11
ISSN0891-5849
DOIs
Publication statusPublished - 15 Mar 2012
Externally publishedYes

    Research areas

  • Animals, Catalytic Domain, Cell Line, Creatine Kinase, Enzyme Activation, Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+), Macrophages, Mice, Oxidative Stress, Peroxidase, Sulfenic Acids, Sulfhydryl Compounds, Thiocyanates

ID: 128975162