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Separation of three P450 isozymes from liver microsomes of gilthead seabream treated with ß‐NF and partial purification of cytochrome P4501A1
Author(s) -
Sen Alaattin,
Arıç Emel
Publication year - 1997
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549700201131
Subject(s) - microsome , isozyme , cytochrome p450 , cytochrome , biochemistry , biology , chromatography , chemistry , enzyme
Three distinct cytochrome P450 isozymes, P4501A1, P4502B and an unidentified P450 isozyme were separated and isolated from ß‐NF‐treated gilthead seabream liver microsomes. Cytochrome P4501A1 was partially purified from ß‐NF‐treated gilthead seabream liver microsomes in the presence of detergents Emulgen 913 and cholate and protease inhibitors using two DEAE‐cellulose, Porapak Q and two hydroxylapatite column chromatographies. The overall yield of purified P4501A1 was 1.2% with respect to microsomal total P450 with a specific content of 3 nmol/mg protein. The purified P4501A1 was characterized with respect to spectral, electrophoretic, biocatalytic and immunochemical properties, which are found to be similar to P4501Als purified from other teleost species such as trout, scup, cod and perch suggesting that the P450 we have purified belongs to CYP1A1.