| Texto completo | |
| Autor(es): |
Queiroz, Raphael F.
;
Vaz, Sandra M.
;
Augusto, Ohara
Número total de Autores: 3
|
| Tipo de documento: | Artigo Científico |
| Fonte: | Biochemical Journal; v. 439, p. 9-pg., 2011-11-01. |
| Resumo | |
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 H2O2, 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 |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |