A New Method for the Purification of Cytochrome-p450 From Human Liver-microsomes
(1992) Biomedical Chromatography — Vol. 6, n° 5, p. 248-250 (1992)
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- Abstract
- 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.
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Isa, M., Cumps, J., Fossoul, C., & Atassi, G. (1992). A New Method for the Purification of Cytochrome-p450 From Human Liver-microsomes. Biomedical Chromatography, 6(5), 248-250. https://doi.org/10.1002/bmc.1130060510 (Original work published 1992)