PT  - JOURNAL ARTICLE
AU  - Tonnessen, Britt H.
AU  - Severson, Sandra R.
AU  - Hurt, Richard D.
AU  - Miller, Virginia M.
TI  - Modulation of Nitric-Oxide Synthase by Nicotine
DP  - 2000 Nov 01
TA  - Journal of Pharmacology and Experimental Therapeutics
PG  - 601--606
VI  - 295
IP  - 2
4099  - http://jpet.aspetjournals.org/content/295/2/601.short
4100  - http://jpet.aspetjournals.org/content/295/2/601.full
SO  - J Pharmacol Exp Ther2000 Nov 01; 295
AB  - Effects of nicotine on arterial endothelium-dependent relaxations mediated by nitric oxide are controversial. Experiments were designed to test the hypothesis that nicotine can directly alter activity of endothelial nitric-oxide synthase (eNOS). NOS from aortic endothelial cells of untreated dogs and recombinant eNOS, neuronal NOS, and inducible NOS were used for these experiments. NOS activity was determined as conversion of l-[3H]arginine tol-[3H]citrulline in the absence or presence of nicotine (10−7–10−3 M) in vitro. In separate assays, concentrations of cofactors NADPH, FAD, and tetrahydrobioprotein were reduced by half to assess for possible interaction with nicotine. With enzyme from aortic endothelial cells, total and calcium-dependent accumulation of citrulline increased by 30% in the presence of 10−5 M nicotine. Nicotine dose dependently also increased citrulline accumulation by recombinant eNOS and neuronal NOS but not inducible NOS. Effects of nicotine on accumulation of citrulline by isolated eNOS and recombinant eNOS were further modulated by changes in the concentration of NADPH in the incubation solution. Our data demonstrate a significant effect of nicotine on eNOS-mediated citrulline accumulation. These results suggest that effects of nicotine on production of nitric oxide may depend on NADPH or oxygen radical interactions with NOS and thus may explain, in part, inconsistent findings of changes in production of endothelium-derived nitric oxide with nicotine administration. The American Society for Pharmacology and Experimental Therapeutics