title: Brain nitric oxide synthase is a biopterin- and flavin-containing multi-functional oxido-reductase.
abstract: Brain nitric oxide synthase is a Ca2+/calmodulin-regulated enzyme which converts L-arginine into NO. Enzymatic activity of this enzyme essentially depends on NADPH and is stimulated by tetrahydrobiopterin (H4biopterin). We found that purified NO synthase contains enzyme-bound H4biopterin, explaining the enzymatic activity observed in the absence of added cofactor. Together with the finding that H4biopterin was effective at substoichiometrical concentrations, these results indicate that NO synthase essentially depends on H4biopterin as a cofactor which is recycled during enzymatic NO formation. We found that the purified enzyme also contains FAD, FMN and non-heme iron in equimolar amounts and exhibits striking activities, including a Ca2+/calmodulin-dependent NADPH oxidase activity, leading to the formation of hydrogen peroxide at suboptimal concentrations of L-arginine or H4biopterin.
authors.affiliation: Institut für Pharmakologie, Freie Universität Berlin, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Mayer
authors.forename: M
authors.initials: M
authors.lastname: John
authors.forename: B
authors.initials: B
authors.lastname: Heinzel
authors.forename: E R
authors.initials: ER
authors.lastname: Werner
authors.forename: H
authors.initials: H
authors.lastname: Wachter
authors.forename: G
authors.initials: G
authors.lastname: Schultz
authors.forename: E
authors.initials: E
authors.lastname: Böhme
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000378:metabolism; D001708:Biopterins / Q000031:analogs & derivatives / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002531:Cerebellum / Q000201:enzymology*; D002956:Citrulline / Q000378:metabolism; D005486:Flavin Mononucleotide / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D009249:NADP / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase; D013053:Spectrophotometry; D013552:Swine
chemical_list: D002147:Calmodulin; D005182:Flavin-Adenine Dinucleotide; D001708:Biopterins; D002956:Citrulline; D009569:Nitric Oxide; D009249:NADP; D005486:Flavin Mononucleotide; D001120:Arginine; D006861:Hydrogen Peroxide; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; C003402:sapropterin; D002118:Calcium