NO synthase isoforms specifically modify peroxynitrite reactivity.
Nitric oxide synthases (NOSs) are multi-domain hemothiolate proteins that are the sole source of nitric oxide (NO) in mammals. NOSs can also be a source or a sink for peroxynitrite (PN), an oxidant that is suspected to be involved in numerous physiopathological processes. In a previous study, we showed that the oxygenase domain of the inducible NOS (iNOSoxy) reacts with PN and changes its oxidative reactivity [Maréchal A, Mattioli TA, Stuehr DJ & Santolini J (2007) J Biol Chem 282, 14101-14112]. Here we report a similar analysis on two other NOS isoforms, neuronal NOS (nNOS) and a bacterial NOS-like protein (bsNOS). All NOSs accelerated PN decomposition, with accumulation of a similar heme intermediate. The kinetics of PN decomposition and heme transitions were comparable among NOSs. However, their effects on PN reactivity differ greatly. All isoforms suppressed PN two-electron oxidative activity, but iNOSoxy enhanced PN one-electron oxidation and nitration potencies, the oxygenase domain of nNOS (nNOSoxy) affected them minimally, and bsNOS abolished all PN reactivities. This led to the loss of both NOS and PN decomposition activities for nNOSoxy and iNOSoxy, which may be linked to the reported alterations in their electronic absorption spectra. Bacterial bsNOS was affected to a lesser extent by reaction with PN. We propose that these differences in PN reactivity among NOSs might arise from subtle differences in their heme pockets, and could reflect the physiological specificity of each NOS isoform, ranging from oxidative stress amplification (iNOS) to detoxification (bsNOS).
Maréchal
Amandine
A
Laboratoire Stress Oxydant et Détoxication, iBiTec-S, CEA Saclay, Gif-sur-Yvette Cedex, France.
Mattioli
Tony A
TA
Stuehr
Dennis J
DJ
Santolini
Jérôme
J
D000818:Animals; D002384:Catalysis; D019281:Dimerization; D004789:Enzyme Activation; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008322:Mammals; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D010084:Oxidation-Reduction; D030421:Peroxynitrous Acid / Q000378:metabolism*; D013379:Substrate Specificity
D007527:Isoenzymes; D030421:Peroxynitrous Acid; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II