Cited by 494Nitric Oxide and Endothelin EffectsRedox biology and oxidative stressNeutrophil, Myeloperoxidase and Oxidative Mechanisms

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

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Zou, Ming-Hui, Shi, Chaomei, Cohen, Richard A.·2002·Journal of Clinical Investigation

DOI 10.1172/jci0214442

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Abstract

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 .-indiabetes, 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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Authors: Zou, M.-H., Shi, C., & Cohen, R. A.
Affiliations: Boston University (US)
Corresponding author: Zou, Ming-Hui
ORCID: Zou, Ming-Hui
Cohen, Richard A.
Venue: Journal of Clinical Investigation
Publisher: American Society for Clinical Investigation
Published: 2002-03-15
Volume: 109
Issue: 6
Pages: 817-826
ISSN: 0021-9738 (ISSN-L)
DOI: 10.1172/jci0214442
OpenAlex: W4254181710
Cited by: 494
References: 48
Citations by year: 2026: 1 · 2025: 10 · 2024: 7 · 2023: 12 · 2022: 19 · 2021: 12 · 2020: 17 · 2019: 15 · 2018: 19 · 2017: 23 · 2016: 15 · 2015: 22 · 2014: 22 · 2013: 22 · 2012: 29
FWCI: 17.56
Type: journal article
Language: en
Open access: bronze
doi.org
Locations: Journal of Clinical Investigation (publishedVersion)
Funders: American Heart Association
National Institutes of Health (PO1 HL)
Topics: Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
Keywords: peroxynitrite, endothelial nitric oxide synthase, superoxide production, tetrahydrobiopterin, disulfide bonds, oxidative stress, diabetes mellitus, endothelial cells, nitric oxide production

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

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