Charnecki J, Putt D, Kim E Y, Kim H
Oxford Biomedical Research, Inc., Rochester Hills, MI 48309, USA.
Biochem Biophys Res Commun. 1995 Nov 22;216(3):1024-33. doi: 10.1006/bbrc.1995.2723.
Rat cytochrome P450 2B1 antipeptide antibodies were produced by immunizing rabbits with a synthetic peptide antigen. The anti-CYP2B1 IgG obtained did not cross-react with CYP2B2, which has 97% identity in primary sequence of CYP2B1. This result demonstrates that a difference of 2 amino acid residues among 12 is sufficient to produce a form-specific antibody. The CYP2B1 antipeptide IgG inhibited pentoxyresorufin O-dealkylase activity of microsomes obtained from phenobarbital-treated rats in a dose-dependent manner, whereas it did not inhibit ethoxyresorufin O-deethylase activity of microsomes obtained from 3-methylcholanthrene-treated rats. These results suggest that the selected amino acid sequence, which coincides with one of the substrate binding sites of Pseudomonas putida CYP101A (P450cam) and one of the putative substrate binding sites of CYP2B2, is located on the surface of the CYP2B1 molecule, as opposed to inside the molecule or in the lipid bilayer of microsomes.
通过用合成肽抗原免疫兔子来制备大鼠细胞色素P450 2B1抗肽抗体。所获得的抗CYP2B1 IgG与CYP2B2没有交叉反应，CYP2B2在CYP2B1的一级序列中有97%的同一性。该结果表明，12个氨基酸残基中有2个氨基酸残基的差异足以产生一种形式特异性抗体。CYP2B1抗肽IgG以剂量依赖性方式抑制从苯巴比妥处理的大鼠获得的微粒体的戊氧基试卤灵O-脱烷基酶活性，而不抑制从3-甲基胆蒽处理的大鼠获得的微粒体的乙氧基试卤灵O-脱乙基酶活性。这些结果表明，所选的氨基酸序列与恶臭假单胞菌CYP101A（P450cam）的底物结合位点之一以及CYP2B2的推定底物结合位点之一一致，位于CYP2B1分子的表面，而不是分子内部或微粒体的脂质双层中。
