title: Mechanistic studies of the isomerization of peroxynitrite to nitrate catalyzed by distal histidine metmyoglobin mutants.
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 (k(cat) = (5.8 +/- 0.1) x 10(6) M(-1) s(-1), at pH 7.5 and 20 degrees C) and H64D metMb (k(cat) = (4.8 +/- 0.1) x 10(6) M(-1) s(-1), at pH 7.5 and 20 degrees C). The pH dependence of the values of k(cat) shows that HOONO is the species which reacts with the heme. In the presence of physiologically relevant concentrations of CO(2) (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 k(cat). 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 k(cat).
authors.affiliation: Laboratorium für Anorganische Chemie, Eidgenössische Technische Hochschule, ETH Hönggerberg, CH-8093 Zürich, Switzerland. herold@inorg.chem.ethz.ch
authors.forename: Susanna
authors.initials: S
authors.lastname: Herold
authors.forename: Shivashankar
authors.initials: S
authors.lastname: Kalinga
authors.forename: Toshitaka
authors.initials: T
authors.lastname: Matsui
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Watanabe
mesh_terms: D000818:Animals; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D005914:Globins / Q000378:metabolism; D006418:Heme / Q000737:chemistry / Q000378:metabolism; D006639:Histidine / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D007536:Isomerism; D007700:Kinetics; D008786:Metmyoglobin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics*; D009566:Nitrates / Q000737:chemistry* / Q000378:metabolism*; D009584:Nitrogen / Q000032:analysis; D030421:Peroxynitrous Acid / Q000737:chemistry* / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism; D014907:Whales
chemical_list: D009566:Nitrates; D008786:Metmyoglobin; D030421:Peroxynitrous Acid; C002744:3-nitrotyrosine; D014443:Tyrosine; D006418:Heme; D006639:Histidine; D005914:Globins; D009584:Nitrogen