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.