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Inhibition of the chlorinating activity of myeloperoxidase by tempol: revisiting the kinetics and mechanisms

Texto completo
Queiroz, Raphael F. [1] ; Vaz, Sandra M. [1] ; Augusto, Ohara [1]
Número total de Autores: 3
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Inst Quim, Dept Bioquim, BR-05513970 Sao Paulo - Brazil
Número total de Afiliações: 1
Tipo de documento: Artigo Científico
Fonte: Biochemical Journal; v. 439, n. 3, p. 423-431, NOV 1 2011.
Citações Web of Science: 22

The nitroxide tempol (4-hydroxy-2,2,6,6-tetramethyl piperidine-1-oxyl) reduces tissue injury in animal models of inflammation by mechanisms that are not completely understood. MPO (myeloperoxidase), which plays a fundamental role in oxidant production by neutrophils, is an important target for anti-inflammatory action. By amplifying the oxidative potential of H(2)O(2), MPO produces hypochlorous acid and radicals through the oxidizing intermediates MPO-I {[}MPO-porphyrin(center dot+)-Fe(IV)=O] and MPO-II {[}MPO-porphyrin-Fe(IV)=O]. Previously, we reported that tempol reacts with MPO-I and MPO-II with second-order rate constants similar to those of tyrosine. However, we noticed that tempol inhibits the chlorinating activity of MPO, in contrast with tyrosine. Thus we studied the inhibition of MPO-mediated taurine chlorination by tempol at pH 7.4 and re-determined the kinetic constants of the reactions of tempol with MPO-I (k = 3.5 x 10(5) M(-1) . s(-1)) and MPO-II, the kinetics of which indicated a binding interaction (K = 2.0 x 10(-5) M; k = 3.6 x 10(-2) s(-1)). Also, we showed that tempol reacts extremely slowly with hypochlorous acid (k = 0.29 and 0.054 M(-1) . s(-1) at pH 5.4 and 7.4 respectively). The results demonstrated that tempol acts mostly as a reversible inhibitor of MPO by trapping it as MPO-II and the MPO-II-tempol complex, which are not within the chlorinating cycle. After turnover, a minor fraction of MPO is irreversibly inactivated, probably due to its reaction with the oxammonium cation resulting from tempol oxidation. Kinetic modelling indicated that taurine reacts with enzyme-bound hypochlorous acid. Our investigation complements a comprehensive study reported while the present study was underway {[}Rees, Bottle, Fairfull-Smith, Malle, Whitelock and Davies (2009) Biochem. J. 421, 79-86]. (AU)

Processo FAPESP: 08/57721-3 - Redoxoma
Beneficiário:Ohara Augusto
Linha de fomento: Auxílio à Pesquisa - Temático