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.
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页码：248 / 250
页数：3
相关论文
共 20 条
  • [1] ANALYTICAL STUDY OF MICROSOMES AND ISOLATED SUBCELLULAR MEMBRANES FROM RAT-LIVER .2. PREPARATION AND COMPOSITION OF MICROSOMAL FRACTION
    AMARCOSTESEC, A
    BEAUFAY, H
    WIBO, M
    THINESSE.D
    FEYTMANS, E
    ROBBI, M
    BERTHET, J
    [J]. JOURNAL OF CELL BIOLOGY, 1974, 61 (01) : 201 - 212
  • [2] PURIFICATION OF NADPH - CYTOCHROME-C (CYTOCHROME-P-450) REDUCTASE FROM HAMSTER LIVER-MICROSOMES BY DETERGENT EXTRACTION AND AFFINITY-CHROMATOGRAPHY
    ARDIES, CM
    LASKER, JM
    BLOSWICK, BP
    LIEBER, CS
    [J]. ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) : 39 - 46
  • [3] PARTIAL-PURIFICATION OF HUMAN-LIVER CYTOCHROME-P-450
    BEAUNE, P
    DANSETTE, P
    FLINOIS, JP
    COLUMELLI, S
    MANSUY, D
    LEROUX, JP
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1979, 88 (03) : 826 - 832
  • [4] PURIFICATION OF A NEW CYTOCHROME-P-450 FROM HUMAN-LIVER MICROSOMES
    BEAUNE, P
    FLINOIS, JP
    KIFFEL, L
    KREMERS, P
    LEROUX, JP
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1985, 840 (03) : 364 - 370
  • [5] BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
  • [6] CONNEY AH, 1968, J BIOL CHEM, V243, P3912
  • [7] BIOGENESIS OF ENDOPLASMIC RETICULUM MEMBRANES .2. SYNTHESIS OF CONSTITUTIVE MICROSOMAL ENZYMES IN DEVELOPING RAT HEPATOCYTE
    DALLNER, G
    SIEKEVITZ, P
    PALADE, GE
    [J]. JOURNAL OF CELL BIOLOGY, 1966, 30 (01) : 97 - +
  • [8] Gelboin H V, 1967, Adv Cancer Res, V10, P1, DOI 10.1016/S0065-230X(08)60076-7
  • [9] DEBRISOQUINE-TYPE POLYMORPHISM OF DRUG OXIDATION - PURIFICATION FROM HUMAN-LIVER OF A CYTOCHROME-P450 ISOZYME WITH HIGH-ACTIVITY FOR BUFURALOL HYDROXYLATION
    GUT, J
    GASSER, R
    DAYER, P
    KRONBACH, T
    CATIN, T
    MEYER, UA
    [J]. FEBS LETTERS, 1984, 173 (02) : 287 - 290
  • [10] ISA M, 1989, J PHARM BELG, V44, P41
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