Purification of nitric oxide synthase from bovine brain: immunological characterization and tissue distribution.
Nitric oxide (NO) synthase (EC 1.14.23) was purified to homogeneity from bovine cerebrum. The molecular weight of NO synthase was estimated to be 150 kDa by both SDS/PAGE and gel filtration at high salt concentration. For activity, the enzyme required NADPH, Ca2+, calmodulin and tetrahydrobiopterin as cofactors. Rabbit polyclonal antibody to bovine brain NO synthase reacted with 150 kDa NO synthase in various bovine and rat organs, including the brain, pituitary and adrenal glands, but not with that in stimulated macrophages, indicating that there are at least two immunologically distinct NO synthases.
Biochemistry Division, National Cancer Center Research Institute, Tokyo, Japan.
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H
Ohshima
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Oguchi
H
H
Adachi
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Iida
H
H
Suzuki
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T
Sugimura
H
H
Esumi
D000594:Amino Acid Oxidoreductases / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification*; D000818:Animals; D001708:Biopterins / Q000031:analogs & derivatives / Q000494:pharmacology; D001921:Brain / Q000201:enzymology* / Q000276:immunology; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D002417:Cattle; D007527:Isoenzymes; D007700:Kinetics; D009249:NADP / Q000494:pharmacology; D019001:Nitric Oxide Synthase; D051381:Rats; D014018:Tissue Distribution
D002147:Calmodulin; D007527:Isoenzymes; D001708:Biopterins; D009249:NADP; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; C003402:sapropterin; D002118:Calcium