Expression of rat neuronal nitric oxide synthase in Saccharomyces cerevisiae.
The rat neuronal nitric oxide synthase (nNOS) cDNA was expressed in Saccharomyces cerevisiae under the control of a hybrid PGK/GAL promoter, PAL. Galactose induction resulted in the production of a soluble 160 kDa protein that was recognised by anti-neuronal NOS antibody. NOS activity was detected by the conversion of [3H]arginine to [3H]citrulline and required the addition of the cofactors, calmodulin and tetrahydrobiopterin. The activity was inhibited by the arginine analogues NG-nitro-L-arginine and NG-nitro-L-arginine methyl ester.
Jones
S
S
Delta Biotechnology Limited, Nottingham, UK.
Tasab
M
M
Ogden
J E
JE
Ballance
D J
DJ
Powell
M J
MJ
D000818:Animals; D001483:Base Sequence; D001709:Biotechnology; D018076:DNA, Complementary / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D009474:Neurons / Q000201:enzymology*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D019001:Nitric Oxide Synthase