Tyrosine nitration by peroxynitrite formed from nitric oxide and superoxide generated by xanthine oxidase

T. Sawa, T. Akaike, H. Maeda

研究成果: Article査読

171 被引用数 (Scopus)

抄録

Peroxynitrite (ONOO-) is a potent nitrating and oxidizing agent that is formed by a rapid reaction of nitric oxide (NO) with superoxide anion (O2-). It appears to be involved in the pathophysiology of many inflammatory and neurodegenerative diseases. It has recently been reported (Pfeiffer, S., and Mayer, B. (1998) J. Biol. Chem. 273, 27280-27285) that ONOO- generated at neutral pH from NO and O2- (NO/O2-) was substantially less efficient than preformed ONOO- at nitrating tyrosine. Here we re-evaluated tyrosine nitration by NO/O2- with a shorter incubation period and a more sensitive electrochemical detection system. Appreciable amounts of nitrotyrosine were produced by ONOO- formed in situ (2.9 μM for 5 min; 10 nM/s) by NO/O2- flux obtained from propylamine NONOate (CH3N[N(O)NO]- (CH2)3NH2+CH3) and xanthine oxidase using pterin as a substrate in phosphate buffer (pH 7.0) containing 0.1 mM L-tyrosine. The yield of nitrotyrosine by this NO/O2- flux was approximately 70% of that produced by the same flux of preformed ONOO- (2.9 μM/5 min). When hypoxanthine was used as a substrate, tyrosine nitration by NO/O2- was largely eliminated because of the inhibitory effect of uric acid produced during the oxidation of hypoxanthine. Tyrosine nitration caused by NO/O2- was inhibited by the ONOO- scavenger ebselen and was enhanced 2-fold by NaHCO3, as would be expected, because CO2 promotes tyrosine nitration. The profile of nitrotyrosine and dityrosine formation produced by NO/O2- flux (2.9 μM/5 min) was consistent with that produced by preformed ONOO-. Tyrosine nitration predominated compared with dityrosine formation caused by a low nanomolar flux of ONOO- at physiological concentrations of free tyrosine (<0.5 mM). In conclusion, our results show that NO generated with O2- nitrates tyrosine with a reactivity and efficacy similar to those of chemically synthesized ONOO-, indicating that ONOO- can be a significant source of tyrosine nitration in physiological and pathological events in vivo.

本文言語English
ページ(範囲)32467-32474
ページ数8
ジャーナルJournal of Biological Chemistry
275
42
DOI
出版ステータスPublished - 2000 10月 20

ASJC Scopus subject areas

  • 生化学
  • 分子生物学
  • 細胞生物学

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