Nitric oxide trapping of the tyrosyl radical of prostaglandin H synthase-2 leads to tyrosine iminoxyl radical and nitrotyrosine formation

被引:151
作者
Gunther, MR
Hsi, LC
Curtis, JF
Gierse, JK
Marnett, LJ
Eling, TE
Mason, RP
机构
[1] NIEHS,NIH,MOL CARCINOGENESIS LAB,RES TRIANGLE PK,NC 27709
[2] GD SEARLE & CO,ST LOUIS,MO 63167
[3] VANDERBILT UNIV,SCH MED,DEPT BIOCHEM,NASHVILLE,TN 37232
关键词
D O I
10.1074/jbc.272.27.17086
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The determination of protein nitrotyrosine content has become a frequently used technique for the detection of oxidative tissue damage, Protein nitration has been suggested to be a final product of the production of highly reactive nitrogen oxide intermediates (e.g. peroxynitrite) formed in reactions between nitric oxide (NO.) and oxygen-derived species such as superoxide. The enzyme prostaglandin H synthase-2 (PHS-2) forms one or more tyrosyl radicals duping its enzymatic catalysis of prostaglandin formation, In the presence of She NO.-generator diethylamine nonoate, the electron spin resonance spectrum of the PHS-a-derived tyrosyl radical is replaced by the spectrum of another free radical containing a nitrogen atom. The magnitude of the nitrogen hyperfine coupling constant in the latter species unambiguously identifies it as an iminoxyl radical, which is likely formed by the oxidation of nitrosotyrosine, a stable product of the addition of NO. to tyrosyl radical. Addition of superoxide dismutase did not alter the spectra, indicating that peroxynitrite was not involved. Western blot analysis of PHS-2 after exposure to the NO.-generator revealed nitrotyrosine formation. The results provide a mechanism for nitric oxide-dependent tyrosine nitration that does not require formation of more highly reactive nitrogen oxide intermediates such as peroxynitrite or nitrogen dioxide.
引用
收藏
页码:17086 / 17090
页数:5
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