Nitric oxide binding to the heme of neuronal nitric-oxide synthase links its activity to changes in oxygen tension

Neuronal nitric-oxide synthase (NOS-1) is a hemeprotein that generates NO and citrulline from L-arginine, O2, and NADPH. During catalysis, a majority of NOS-1 binds self-generated NO and converts to a ferrous-NO complex, which causes it to operate at a fraction of its maximum possible activity during the steady state (Abu-Soud, H. M., Wang, J., Rousseau, D. L., Fukuto, J., Ignarro, L. J., and Stuehr, D. J. (1995) J. Biol. Chem. 270, 22997-23006). To examine how NO complex formation affects the O2 response of NOS-1, we measured rates of NO synthesis and NADPH oxidation versus O2 concentration in the presence and absence of L-arginine. In the absence of L-arginine, NOS-1 catalyzed simple O2 reduction, and its heme iron displayed a typical affinity for O2 (estimated KmO2