PT  - JOURNAL ARTICLE
AU  - J A Mitchell
AU  - K L Kohlhaas
AU  - T Matsumoto
AU  - J S Pollock
AU  - U Förstermann
AU  - T D Warner
AU  - H H Schmidt
AU  - F Murad
TI  - Induction of NADPH-dependent diaphorase and nitric oxide synthase activity in aortic smooth muscle and cultured macrophages.
DP  - 1992 Jun 01
TA  - Molecular Pharmacology
PG  - 1163--1168
VI  - 41
IP  - 6
4099  - http://molpharm.aspetjournals.org/content/41/6/1163.short
4100  - http://molpharm.aspetjournals.org/content/41/6/1163.full
SO  - Mol Pharmacol1992 Jun 01; 41
AB  - Lipopolysaccharide (LPS), either alone or in combination with cytokines, induces nitric oxide (NO) synthase activity in cells that normally release little or no NO. In arterial smooth muscle cells and various macrophage cell lines, NO synthase activity is induced after several hours of incubation with LPS. In brain, NADPH-dependent diaphorase activity has been associated with constitutive NO synthase. Here we show that incubation of rat aorta or cultured macrophages with LPS causes a time-dependent induction of NO synthase. The NO synthase activity in both rat aorta and macrophages was calcium independent and inhibited by NG-monomethyl-L-arginine and NG-nitro-L-arginine. We also found that LPS caused a time-dependent induction in NADPH-dependent diaphorase activity in both rat aorta and cultured macrophages. The diaphorase activity was mainly NADPH dependent and NADH independent. NO synthase activity and NADPH-diaphorase activity in crude cytosol from LPS-treated macrophages were found to co-purify, using 2',5'-ADP-Sepharose followed by Superose-6 gel permeation chromatography.