title: Enzymatic formation of nitrogen oxides from L-arginine in bovine brain cytosol.
abstract: In dialyzed bovine brain cytosol, the enzymatic formation of nitrogen oxides was directly determined. The basal formation of nitrite and nitrate was concentration-dependently enhanced by L-arginine (EC50 about 3.10(-5) M). Both the basal and L-arginine induced formations were inhibited by NG-monomethyl-L-arginine (EC50 about 2.10(-4) M). In the presence of L-arginine, a concomitant formation of citrulline was detected. L-Arginine methyl ester also served as a substrate, but neither D-arginine, D-arginine methyl ester nor N alpha-benzoyl-L-arginine ethyl ester did so. The formation of nitrite and nitrate was time-dependent, increased linearly with the protein concentration of the cytosol and was not observed when the cytosolic proteins were heat-denaturated. Exogenous NADPH (or NADP+) concentration-dependently enhanced the formation of nitrite and nitrate, whereas NADH, NAD+, FAD, Ca2+, Mg2+ and calmodulin were ineffective. These results indicate that bovine brain contains a cytosolic enzyme which uses NADPH or NADP+ as cofactors to form nitrogen oxides from both an endogenous non-dialyzable substrate and from L-arginine.
authors.affiliation: Institut für Pharmakologie, Freie Universität Berlin, F.R.G.
authors.forename: H H
authors.initials: HH
authors.lastname: Schmidt
authors.forename: P
authors.initials: P
authors.lastname: Wilke
authors.forename: B
authors.initials: B
authors.lastname: Evers
authors.forename: E
authors.initials: E
authors.lastname: Böhme
mesh_terms: D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000378:metabolism* / Q000494:pharmacology; D001921:Brain / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D003600:Cytosol / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000494:pharmacology; D007700:Kinetics; D009243:NAD / Q000494:pharmacology; D009249:NADP / Q000494:pharmacology; D009589:Nitrogen Oxides / Q000378:metabolism*; D010084:Oxidation-Reduction
chemical_list: D009589:Nitrogen Oxides; D009243:NAD; D005182:Flavin-Adenine Dinucleotide; D009249:NADP; D001120:Arginine; D002118:Calcium