Oxidation of the zinc-thiolate complex and uncoupling of endothelial nitric oxide synthase by peroxynitrite

被引:547
作者
Zou, MH [1 ]
Shi, CM [1 ]
Cohen, RA [1 ]
机构
[1] Boston Univ, Sch Med, Vasc Biol Unit X708, Dept Med,Whitaker Cardiovasc Inst, Boston, MA 02118 USA
关键词
D O I
10.1172/JCI200214442
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Nitric oxide (NO) is produced by NO synthase (NOS) in many cells and plays important roles in the neuronal, muscular, cardiovascular, and immune systems. In various disease conditions, all three types of NOS (neuronal, inducible, and endothelial) are reported to generate oxidants through unknown mechanisms. We present here the First evidence that peroxynitrite (ONOO-) releases zinc from the zinc-thiolate cluster of endothelial NOS (eNOS) and presumably forms disulfide bonds between the monomers. As a result, disruption of the otherwise SDS-resistant eNOS dimers occurs under reducing conditions. eNOS catalytic activity is exquisitely sensitive to ONOO-, which decreases NO synthesis and increases superoxide anion (O-2(-)) production by the enzyme. The reducing cofactor tetrahydrobiopterin is not oxidized, nor does it prevent oxidation of eNOS by the same low concentrations of OONO-. Furthermore, eNOS derived from endothelial cells exposed to elevated glucose produces more O-2(-), and, like eNOS purified from diabetic LDL receptor-deficient mice, contains less zinc and fewer SDS-resistant dimers. Hence, eNOS exposure to oxidants including ONOO- causes increased enzymatic uncoupling and generation of O-2(-) in diabetes, contributing further to endothelial cell oxidant stress. Regulation of the zinc-thiolate center of NOS by ONOO- provides a novel mechanism for modulation of the enzyme function in disease.
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页码:817 / 826
页数:10
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