PMID: 2127060Jul 1, 1990

Purification of cytochrome P-450 and NADPH cytochrome p-450 reductase from human liver

Journal de pharmacie de Belgique
M IsaG Atassi


Two methods for the purification of cytochromes-P450 from microsomes of human liver are described. Method A: Cyt-P450 were solubilized from microsomes using a non ionic detergent, the Lubrol. The Cyt-P450 were purified by affinity, hydrophobicity followed by ion-exchange chromatography on DEAE-5PW column (HPLC) with an overall yield of 18% and a specific activity of 10 nmole/mg of protein. The recovery of NADPH Cyt-P450 reductase by method A (affinity) is about 60% with a specific activity of 16.2 U.I./mg of protein. Method B: Cyt-P450 were solubilized from microsomes using a zwitterionic detergent, the CHAPS. Cyt-P450 were filtered and separated by chromatofocusing on Mono-P column (HPLC). By this method it was possible to increase strongly the specific activity keeping a yield of 50% of Cyt-P450. Also it was possible to apply this method to small samples of human liver like biopsies (0.5 to 2.5 g).

Related Concepts

NADPH-Ferrihemoprotein Reductase
Cytochrome P450
Cytochrome P-450 Oxygenase
Microsomes, Liver
High Pressure Liquid Chromatography Procedure
Purification Aspects
CYP2B6 protein, human

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