Mouse liver testosterone 15 alpha-hydroxylase (cytochrome P-450(15) alpha). Purification, regioselectivity, stereospecificity, and sex-dependent expression.
作者： N Harada , M Negishi
DOI: 10.1016/S0021-9258(17)43598-8
关键词:
摘要: Testosterone 15 alpha-hydroxylase (cytochrome P-450(15) alpha) was purified from female 129/J mouse liver microsomes based on its specific activities in the eluates columns of octylamino-Sepharose 4B, hydroxylapatite, DEAE-Bio-Gel A, and CM52 chromatography. The alpha-hydroxylation activity five times higher than male microsomes. cytochrome P-450 content alpha fraction 14.5 nmol/mg protein. Soret peak reduced P-450-CO complex 451 nm. apparent subunit molecular weight 48,000, protein appeared as only one major band sodium dodecyl sulfate-polyacrylamide gels. testosterone reconstituted with 94 nmol/min/nmol 1349 nmol/min/mg protein, these were about 65- 1000-fold higher, respectively, solubilized exhibited high regioselectivity stereospecificity for hydroxylation. More 95% metabolites formed by alpha-hydroxytestosterone. Virtually 100% microsomal can be accounted alpha. able to catalyze benzphetamine N-demethylation, 7-ethoxycoumarin O-de-ethylation, aniline 4-hydroxylation, benzo(alpha)pyrene 3-hydroxylation, acetanilide lauric acid (11 + 12)-hydroxylation at various turnover rates, indicating broad substrate specificity oxidations xenobiotics. This is sharp contrast
sci-hub.st 参考文章(27)
Jacob V. Maizel, Polyacrylamide Gel Electrophoresis of Viral Proteins Methods in Virology. ,vol. 5, pp. 179- 246 ,(1971) , 10.1016/B978-0-12-470205-9.50011-3
D.E. Ryan, P.E. Thomas, W. Levin, Hepatic microsomal cytochrome P-450 from rats treated with isosafrole. Purification and characterization of four enzymic forms. Journal of Biological Chemistry. ,vol. 255, pp. 7941- 7955 ,(1980) , 10.1016/S0021-9258(19)43925-2
Kenneth W. McKerns, Steroid hormones and metabolism ,(1969)
Pedro Cuatrecasas, Christian B. Anfinsen, [31] Affinity chromatography Methods in Enzymology. ,vol. 22, pp. 345- 378 ,(1971) , 10.1016/0076-6879(71)22033-4
D W Nebert, H V Gelboin, Substrate-inducible Microsomal Aryl Hydroxylase in Mammalian Cell Culture: I. Assay and properties of induced enzyme Journal of Biological Chemistry. ,vol. 243, pp. 6242- 6249 ,(1968) , 10.1016/S0021-9258(18)94484-4
E.F. Johnson, Isolation and characterization of a constitutive form of rabbit liver microsomal cytochrome P-450. Journal of Biological Chemistry. ,vol. 255, pp. 304- 309 ,(1980) , 10.1016/S0021-9258(19)86298-1
B S Masters, Y Yasukochi, Some properties of a detergent-solubilized NADPH-cytochrome c(cytochrome P-450) reductase purified by biospecific affinity chromatography. Journal of Biological Chemistry. ,vol. 251, pp. 5337- 5344 ,(1976) , 10.1016/S0021-9258(17)33166-6
Jan-Åke Gustafsson, Magnus Ingelman-Sundberg, Regulation and properties of a sex-specific hydroxylase system in female rat liver microsomes active on steroid sulfates. I. General characteristics. Journal of Biological Chemistry. ,vol. 249, pp. 1940- 1945 ,(1974) , 10.1016/S0021-9258(19)42875-5
M. Agosin, A. Morello, R. White, Y. Repetto, J. Pedemonte, Multiple forms of noninduced rat liver cytochrome P-450. Metabolism of 1-(4'-ethylphenoxy)-3,7-dimethyl-6,7-epoxy-trans-2-octene by reconstituted preparations. Journal of Biological Chemistry. ,vol. 254, pp. 9915- 9920 ,(1979) , 10.1016/S0021-9258(19)83604-9
A Y H Lu, M J Coon, Role of Hemoprotein P-450 in Fatty Acid ω-Hydroxylation in a Soluble Enzyme System from Liver Microsomes Journal of Biological Chemistry. ,vol. 243, pp. 1331- 1332 ,(1968) , 10.1016/S0021-9258(19)56992-7