Separation of three P450 isozymes from liver microsomes of gilthead seabream treated with D-NF and partial purification of cytochrome P4501A1


Sen A., Arinc E.

BIOCHEMISTRY AND MOLECULAR BIOLOGY INTERNATIONAL, cilt.41, ss.131-141, 1997 (SCI İndekslerine Giren Dergi) identifier

  • Cilt numarası: 41 Konu: 1
  • Basım Tarihi: 1997
  • Dergi Adı: BIOCHEMISTRY AND MOLECULAR BIOLOGY INTERNATIONAL
  • Sayfa Sayıları: ss.131-141

Özet

Three distinct cytochrome P450 isozymes, P4501A1, P4502B and an unidentified P450 isozyme were separated and isolated from beta-NF-treated gilthead seabream liver microsomes. Cytochrome P4501A1 was partially purified from beta-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 P4501A1s purified from other teleost species such as trout, scup, cod and perch suggesting that the P450 we have purified belongs to CYP1A1.