title: NADPH and heme redox modulate pulmonary artery relaxation and guanylate cyclase activation by NO.
abstract: The hemoprotein oxidant ferricyanide (FeCN) converts the iron of the heme on soluble guanylate cyclase (sGC) from Fe(2+) to Fe(3+), which prevents nitric oxide (NO) from binding the heme and stimulating sGC activity. This study uses FeCN to examine whether modulation of the redox status of the heme on sGC influences the relaxation of endothelium-removed bovine pulmonary arteries (BPA) to NO. Pretreatment of the homogenate of BPA with 50 microM FeCN resulted in a loss of stimulation of sGC activity by the NO donor 10 microM S-nitroso-N-acetylpenicillamine (SNAP). In the FeCN-treated homogenate reconcentrated to the enzyme levels in BPA, 100 microM NADPH restored NO stimulation of sGC, and this effect of NADPH was prevented by an inhibitor of flavoprotein electron transport, 1 microM diphenyliodonium (DPI). In BPA the relaxation to SNAP was not altered by FeCN, inhibitors of NADPH generation by the pentose phosphate pathway [250 microM 6-aminonicotinamide (6-AN) and 100 microM epiandrosterone (Epi)], or 1 microM DPI. However, the combination of FeCN with 6-AN, Epi, or DPI inhibited (P < 0.05) relaxation to SNAP without significantly altering the relaxation of BPA to forskolin. The inhibitory effects of 1 microM 1H-[1,2, 4]oxadiazolo[4,3-a]quinoxalin-1-one (a probe that appears to convert NO-heme of sGC to its Fe(3+)-heme form) on relaxation to SNAP were also enhanced by DPI. These observations suggest that a flavoprotein containing NADPH oxidoreductase may influence cGMP-mediated relaxation of BPA to NO by maintaining the heme of sGC in its Fe(2+) oxidation state.
authors.affiliation: Department of Physiology, New York Medical College, Valhalla, New York 10595, USA.
authors.forename: S A
authors.initials: SA
authors.lastname: Gupte
authors.forename: T
authors.initials: T
authors.lastname: Rupawalla
authors.forename: D
authors.initials: D
authors.lastname: Phillibert
authors.forename: M S
authors.initials: MS
authors.lastname: Wolin
mesh_terms: D000818:Animals; D002417:Cattle; D005576:Colforsin / Q000494:pharmacology; D006152:Cyclic GMP / Q000378:metabolism; D004579:Electron Transport / Q000502:physiology; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005292:Ferricyanides / Q000494:pharmacology; D005420:Flavoproteins / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D066298:In Vitro Techniques; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D010069:Oxadiazoles / Q000494:pharmacology; D010084:Oxidation-Reduction; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D010427:Pentose Phosphate Pathway / Q000502:physiology; D011651:Pulmonary Artery / Q000187:drug effects / Q000201:enzymology*; D011810:Quinoxalines / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: C095284:1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one; D004791:Enzyme Inhibitors; D005292:Ferricyanides; D005420:Flavoproteins; D010069:Oxadiazoles; D011810:Quinoxalines; D017382:Reactive Oxygen Species; C110881:S-nitro-N-acetylpenicillamine; D013438:Sulfhydryl Compounds; C007931:hexacyanoferrate III; D005576:Colforsin; D009569:Nitric Oxide; D006418:Heme; D009249:NADP; D009247:NADH, NADPH Oxidoreductases; D006162:Guanylate Cyclase; D005978:Glutathione; D010396:Penicillamine; D006152:Cyclic GMP