title: Nitric oxide synthases in neuronal cells, macrophages and endothelium are NADPH diaphorases, but represent only a fraction of total cellular NADPH diaphorase activity.
abstract: NADPH diaphorase activity is used as a histochemical marker for neuronal nitric oxide (NO) synthase; however, it remains unclear whether these activities are directly correlated in all tissues. In N1E-115 neuroblastoma cells, NADPH diaphorase activity was found primarily in the particulate fraction, whereas NO synthase activity was mostly soluble. Non-induced macrophages expressed significant NADPH diaphorase activity (which was mostly particulate) but virtually no NO synthase activity. Induction of macrophages produced marked increases in both NO synthase and NADPH diaphorase activities in the soluble and particulate fractions. In endothelial cells, both NO synthase and NADPH diaphorase activities were found mostly in the particulate fraction. Purified NO synthases from brain (type I), macrophages (type II), and endothelium (type III) all showed NADPH diaphorase activity; relative activities were: macrophage > endothelium > brain. These data indicate that all known NO synthases are NADPH diaphorases; however, NO synthases represent only a fraction of total cellular NADPH diaphorase activity and these activities are not always co-localized.
authors.affiliation: Vascular Biology Project, Abbott Laboratories, IL 60064.
authors.forename: W R
authors.initials: WR
authors.lastname: Tracey
authors.forename: M
authors.initials: M
authors.lastname: Nakane
authors.forename: J S
authors.initials: JS
authors.lastname: Pollock
authors.forename: U
authors.initials: U
authors.lastname: Förstermann
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D000818:Animals; D001011:Aorta; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D003600:Cytosol / Q000201:enzymology; D004730:Endothelium, Vascular / Q000201:enzymology*; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008264:Macrophages / Q000201:enzymology*; D051379:Mice; D009252:NADPH Dehydrogenase / Q000378:metabolism*; D009447:Neuroblastoma; D009474:Neurons / Q000201:enzymology*; D019001:Nitric Oxide Synthase; D013347:Subcellular Fractions / Q000201:enzymology; D014407:Tumor Cells, Cultured
chemical_list: D007527:Isoenzymes; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D009252:NADPH Dehydrogenase