Mechanistic studies of the isomerization of peroxynitrite to nitrate catalyzed by distal histidine metmyoglobin mutants

Susanna Herold, Shivashankar Kalinga, Toshitaka Matsui, Yoshihito Watanabe

Research output: Contribution to journalArticlepeer-review

44 Citations (Scopus)

Abstract

Hemoproteins are known to react with the strong nitrating and oxidizing agent peroxynitrite according to different mechanisms. In this article, we show that the iron(III) forms of the sperm whale myoglobin (sw Mb) mutants H64A, H64D, H64L, F43W/H64L, and H64Y/H93G catalyze the isomerization of peroxynitrite to nitrate. The two most efficient catalysts are H64A (kcat = (5.8 ± 0.1) × 106 M-1 s-1, at pH 7.5 and 20 °C) and H64D metMb (kcat = (4.8 ± 0.1) × 106 M-1 s-1, at pH 7.5 and 20 °C). The pH dependence of the values of kcat shows that HOONO is the species which reacts with the heme. In the presence of physiologically relevant concentrations of CO2 (1.2 mM), the decay of peroxynitrite is accelerated by these metMb mutants via the concurring reaction of HOONO with their iron(III) centers. Studies in the presence of free added tyrosine show that the metMb mutants prevent peroxynitrite-mediated nitration. The efficiency of the different sw metMb mutants correlates with the value of kcat. Finally, we show that sw WT-metMb is nitrated to a larger extent than horse heart metMb, a result that suggests that the additional Tyr151 is a site of preferential nitration. Again, the extent of nitration of the tyrosine residues of the metMb mutants correlates with the values of kcat.

Original languageEnglish
Pages (from-to)6945-6955
Number of pages11
JournalJournal of the American Chemical Society
Volume126
Issue number22
DOIs
Publication statusPublished - 2004 Jun 9

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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