Evidence for the presence of cytochrome P-450 functional in lanosterol 14 alpha-demethylation in microsomes of aerobically grown respiring yeast.
Recent studies have shown that a cytochrome P-450 present in microsomes of semi-anaerobically grown cells of Saccharomyces cerevisiae is functional in the 14 alpha-demethylation of lanosterol (4,4,14 alpha-trimethyl-5 alpha-cholesta-8,24-dien-3 beta-ol), but the occurrence of the same cytochrome P-450 in microsomes of aerobically grown yeast cells has not yet been reported. In this study, the microsomal fraction from aerobically grown cells was found to catalyze the lanosterol demethylation in the presence of NADPH and O2 and that this activity was sensitive to CO. In Ouchterlony double diffusion test, antibodies to the yeast cytochrome P-450 formed a single precipitin line with the microsomal fraction as well as with the purified yeast cytochrome P-450 and the two precipitin lines fused with each other. Furthermore, the antibodies inhibited the lanosterol demethylation activity of the microsomal fraction from aerobically grown cells. The quadratic-derivative absorption spectrum of the microsomal fraction measured in the presence of both Na2S2O4 and CO showed an absorption band at 450 nm which is attributable to the reduced CO compound of cytochrome P-450. These facts led to the conclusion that cytochrome P-450 actually exists in aerobically grown yeast and participates in the lanosterol 14 alpha-demethylation which is essential for the ergosterol (5 alpha-ergosta-5,7,22-trien-3 beta-ol) biogenesis by yeast.
D000332:Aerobiosis; D000936:Antigen-Antibody Complex; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005779:Immunodiffusion; D007700:Kinetics; D007810:Lanosterol / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism*; D010101:Oxygen Consumption; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013053:Spectrophotometry; D005719:gamma-Globulins
D000936:Antigen-Antibody Complex; D005719:gamma-Globulins; D007810:Lanosterol; D003577:Cytochrome P-450 Enzyme System