A NEW METHOD FOR THE PURIFICATION OF CYTOCHROME-P450 FROM HUMAN LIVER-MICROSOMES

被引：3
|
作者：
ISA, M [1 ]
CUMPS, J [1 ]
FOSSOUL, C [1 ]
ATASSI, G [1 ]
机构：
[1] CATHOLIC UNIV LOUVAIN,ECOLE PHARM,CHIM PHARMACEUT LAB,B-1348 LOUVAIN,BELGIUM
来源：
BIOMEDICAL CHROMATOGRAPHY | 1992年 / 6卷 / 05期
关键词：
D O I：
10.1002/bmc.1130060510
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A procedure for the solubilization and purification of cytochrome-P450 (cyt-P450) from human liver microsomes is described. Successive treatment of microsomes with protease XXVII and 3-(3-cholamidopropyl)dimethylammoniopropanesulphonic acid gave a solubilized cyt-P450 in more than 80% yield and with a three-fold increae in specific activity. With this treatment it was possible to eliminate 80% of cytochrome-b5 and 75% of NADPH cyt-P450 reductase. The solubilized cyt-P450 was filtered on a Sephacryl-200 column and then subjected to high performance liquid chromatography with a Mono-P column (chromatofocussing). The recovery of separated cyt-P450 was about 50% with a specific activity of 11.5 nmol cyt-P450/mg protein. Also with this technique it was possible to determinate the isoelectric points of cyt-P450. These results allowed us to confirm the usefulness of our method, for the study the cyt-P450 from surgical biopsies.
引用
收藏
页码：248 / 250
页数：3
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