par Isa, Majed 
;Cumps, Jean;Fossoul, Claudine ;Atassi, Ghanem
Référence BMC. Biomedical chromatography, 6, 5, page (248-250)
Publication Publié, 1992-10
;Cumps, Jean;Fossoul, Claudine ;Atassi, Ghanem Référence BMC. Biomedical chromatography, 6, 5, page (248-250)
Publication Publié, 1992-10
Article révisé par les pairs
| Résumé : | 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. |
