Myeloperoxidase (MPO) is the most abundant protein in neutrophils and plays a central role in microbial killing and inflammatory tissue damage. Because most of the non-steroidal anti-inflammatory drugs and other drugs contain a thiol group, it is necessary to understand how these substrates are oxidized by MPO. We have performed transient kinetic measurements to study the oxidation of 14 aliphatic and aromatic mono- and dithiols by the MPO intermediates, Compound I (k3) and Compound II (k4), using sequential mixing stopped-flow techniques. The one-electron reduction of Compound I by aromatic thiols (e.g. methimidazole, 2-mercaptopurine and 6-mercaptopurine) varied by less than a factor of seven (between 1.39±0.12x105 M-1 s-1 and 9.16±1.63x105 M-1 s-1), whereas reduction by aliphatic thiols was demonstrated to depend on their overall net charge and hydrophobic character and not on the percentage of thiol deprotonation or redox potential. Cysteamine, cysteine methyl ester, cysteine ethyl ester and α-lipoic acid showed k3 values comparable to aromatic thiols, whereas a free carboxy group (e.g. cysteine, N-acetylcysteine, glutathione) diminished k3 dramatically. The one-electron reduction of Compound II was far more constrained by the nature of the substrate. Reduction by methimidazole, 2-mercaptopurine and 6-mercaptopurine showed second-order rate constants (k4) of 1.33±0.08x105 M-1 s-1, 5.25±0.07x105 M-1 s-1 and 3.03±0.07x103 M-1 s-1. Even at high concentrations cysteine, penicillamine and glutathione could not reduce Compound II, whereas cysteamine (4.27±0.05x103 M-1 s-1), cysteine methyl ester (8.14±0.08x103 M-1 s-1), cysteine ethyl ester (3.76±0.17x103 M-1 s-1) and α-lipoic acid (4.78±0.07x104 M-1 s-1) were demonstrated to reduce Compound II and thus could be expected to be oxidized by MPO without co-substrates. Copyright (C) 1999 Federation of European Biochemical Societies.
Burner, U., Jantschko, W., & Obinger, C. (1999). Kinetics of oxidation of aliphatic and aromatic thiols by myeloperoxidase compounds I and II. FEBS Letters, 443(3), 290–296. https://doi.org/10.1016/S0014-5793(98)01727-X