Premium
Constitutive nitric oxide synthase in Saccharomyces cerevisiae
Author(s) -
Kanadia R. N.,
Kuo W. N.,
Mcnabb M.,
Botchway A.
Publication year - 1998
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549800203302
Subject(s) - saccharomyces cerevisiae , citrulline , nitric oxide synthase , extracellular , biochemistry , nitric oxide , microbiology and biotechnology , arginine , yeast , western blot , chemistry , blot , intracellular , calmodulin , enzyme , nitrite , biology , nitrate , gene , organic chemistry , amino acid
After removing nonspecific immunoreactivities from crude extract by immunoaffinity chromatography, an immunoreactive‐band at 60kDa of constitutive nitric oxide synthase (cNOS) from Saccharomyces cerevisiae was detected by Western blot using mouse monoclonal anti‐neuronal NOS (cNOS). The activity of yeast cNOS, which was prepared by either histone‐agarose chromatography or anti‐neuronal NOS immunoprecipitation, was monitored by the formation of citrulline. Yeast cNOS was activated in the presence of calmodulin and arginine, whereas it was inhibited by L‐NAME, a mammalian NOS inhibitor. Moreover, actinomycin‐D decreased the extracellular and the intracellular levels of nitrate and nitrite which had been converted from NO. The results suggest that cNOS occurs in unicellular eukaryotes and the enzyme activity can be regulated.