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

TY - JOUR

T1 - Inactivation of thiol-dependent enzymes by hypothiocyanous acid

T2 - role of sulfenyl thiocyanate and sulfenic acid intermediates

AU - Barrett, Tessa J

AU - Pattison, David I

AU - Leonard, Stephen E

AU - Carroll, Kate S

AU - Davies, Michael Jonathan

AU - Hawkins, Clare Louise

N1 - Copyright © 2012 Elsevier Inc. All rights reserved.

PY - 2012/3/15

Y1 - 2012/3/15

N2 - 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.

AB - 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.

KW - Animals

KW - Catalytic Domain

KW - Cell Line

KW - Creatine Kinase

KW - Enzyme Activation

KW - Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)

KW - Macrophages

KW - Mice

KW - Oxidative Stress

KW - Peroxidase

KW - Sulfenic Acids

KW - Sulfhydryl Compounds

KW - Thiocyanates

U2 - 10.1016/j.freeradbiomed.2011.12.024

DO - 10.1016/j.freeradbiomed.2011.12.024

M3 - Journal article

C2 - 22248862

SN - 0891-5849

VL - 52

SP - 1075

EP - 1085

JO - Free Radical Biology & Medicine

JF - Free Radical Biology & Medicine

IS - 6

ER -