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  • Title: Kinetics of the reactions of nitrogen dioxide with glutathione, cysteine, and uric acid at physiological pH.
    Author: Ford E, Hughes MN, Wardman P.
    Journal: Free Radic Biol Med; 2002 Jun 15; 32(12):1314-23. PubMed ID: 12057769.
    Abstract:
    Nitrogen dioxide (NO(2)(*)) is a key biological oxidant. It can be derived from peroxynitrite via the interaction of nitric oxide with superoxide, from nitrite with peroxidases, or from autoxidation of nitric oxide. In this study, submicromolar concentrations of NO(2)(*) were generated in < 1 micros using pulse radiolysis, and the kinetics of scavenging NO(2)(*) by glutathione, cysteine, or uric acid were monitored by spectrophotometry. The formation of the urate radical was observed directly, while the production of the oxidizing radical obtained on reaction of NO(2)(*) with the thiols (the thiyl radical) was monitored via oxidation of 2,2'-azino-bis-(3-ethylthiazoline-6-sulfonic acid). At pH 7.4, rate constants for reaction of NO(2)(*) with glutathione, cysteine, and urate were estimated as approximately 2 x 10(7), 5 x 10(7), and 2 x 10(7) M(-1) s(-1), respectively. The variation of these rate constants with pH indicated that thiolate reacted much faster than undissociated thiol. The dissociation of urate also accelerated reaction with NO(2)(*) at pH > 8. The thiyl radical from GSH reacted with urate with a rate constant of approximately 3 x 10(7) M(-1) s(-1). The implications of these values are: (i) the lifetime of NO(2)(*) in cytosol is < 10 micros; (ii) thiols are the dominant 'sink' for NO(2)(*) in cells/tissue, whereas urate is also a major scavenger in plasma; (iii) the diffusion distance of NO(2)(*) is approximately 0.2 microm in the cytoplasm and < 0.8 microm in plasma; (iv) urate protects GSH against depletion on oxidative challenge from NO(2)(*); and (v) reactions between NO(2)(*) and thiols/urate severely limit the likelihood of reaction of NO(2)(*) with NO* to form N(2)O(3) in the cytoplasm.
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