ANALYTICAL FRACTIONATION OF MICROSOMAL CYTOCHROME-P-450 ISOENZYMES FROM RAT-LIVER BY HIGH-PERFORMANCE ION-EXCHANGE CHROMATOGRAPHY

被引：12
|
作者：
ROOS, PH
机构：
来源：
JOURNAL OF CHROMATOGRAPHY | 1990年 / 521卷 / 02期
关键词：
D O I：
10.1016/0021-9673(90)85050-6
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
Ion-exchange Fast Protein Liquid Chromatography (FPLC) on Mono Q and Mono S was optimized for the analytical separation of microsomal cytochrome P-450 species from rat liver. The effects of detergent, pH, gradient profile and column load on resolution are demonstrated. Successive application of anion- and cation-exchange chromatography leads to eleven separated P-450 fractions. The altered microsomal P450 pattern after treatment of rats with various inducers is reflected by distinct elution profiles. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and enzymatic analysis imply that several FPLC fractions contain more than one P-450 species. Preliminary results are presented showing the suitability of immobilized metal affinity chromatography (IMAC) for general P-450 fractionation and thus for the further resolution of Mono Q and Mono S fractions. Scale-up for preparative P-450 fractionation is easily done by adapting the optimized analytical FPLC procedures to Q- and S-Sepharose Fast Flow. © 1990.
引用
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页码：251 / 265
页数：15
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