STRUCTURAL-ANALYSIS OF THE INTERACTION BETWEEN THE SIDE-CHAIN OF SUBSTRATES AND THE ACTIVE-SITE OF LANOSTEROL 14-ALPHA-DEMETHYLASE (P-450(14DM)) OF YEAST

被引:20
作者
AOYAMA, Y
YOSHIDA, Y
SONODA, Y
SATO, Y
机构
[1] MUKOGAWA WOMENS UNIV,FAC PHARMACEUT SCI,11-68 KOSHIEN KYUBAN CHO,NISHINOMIYA,HYOGO 663,JAPAN
[2] KYORITSU COLL PHARMACEUT SCI,MINATO KU,TOKYO,JAPAN
关键词
CYTOCHROME-P-450 SUBSTRATE RECOGNITION; LANOSTEROL SIDE CHAIN; LANOSTEROL; 14-ALPHA-DEMETHYLASE; LANOSTEROL DERIVATIVE; (YEAST MICROSOME);
D O I
10.1016/0167-4838(92)90400-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The role of the side-chain of lanosterol in the enzyme-substrate interaction of yeast P-450(14DM) (lanosterol 14-alpha-demethylase) was analyzed with lanosterol derivatives having functional groups on the side-chain. Purified P-450(14DM) from Saccharomyces cerevisiae catalyzed 14-alpha-demethylation of 26-hydroxylanosterol and 25-hydroxy-24,25-dihydrolanosterol with a lower activity than lanosterol and 24,25-dihydrolanosterol. This enzyme demethylated the (Z)-24-ethylidene-24,25-dihydrolanosterol with a low rate, but did not metabolize the E-isomer. The apparent K(m) of 26-hydroxylanosterol was 10.8-mu-M, which was higher than that of lanosterol, but lower than that of 24,25-dihydrolanosterol. On the other hand, competition experiments suggested that the affinity of 25-hydroxy-24,25-dihydrolanosterol and (Z)-24-ethylidene-24,25-dihydrolanosterol for P-450(14DM) was significantly lower than that of 24,25-dihydrolanosterol. Integration of the present results with the preceding ones (Aoyama, Y., Yoshida, Y., Sonoda, Y. and Sato, Y. (1991) Biochim. Biophys. Acta, 1081, 262-266 and Aoyama, Y. and Yoshida, Y. (1991) Biochem. Biophys. Res. Commun., 178, 1064-1071) suggests that yeast P-450(14DM) recognizes two parts of the side-chain, the structure around C-24 and the terminal fork consisting of C-25, C-26 and C-27.
引用
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页码:251 / 255
页数:5
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