Mäenpää J, Rane A, Raunio H, Honkakoski P, Pelkonen O
Department of Pharmacology and Toxicology, University of Oulu, Finland.
Biochem Pharmacol. 1993 Feb 24;45(4):899-907. doi: 10.1016/0006-2952(93)90175-v.
Four polyclonal antibodies raised against purified mouse liver cytochrome P450s representing Cyp1a, Cyp2a, Cyp2b and Cyp2c subfamilies were used to detect their related forms in human adult and fetal tissues. In immunoblot analysis, anti-Cyp2c antibody detected two to three proteins in adult livers and one to three proteins in 70% of the 18 fetal livers studied. Anti-Cyp2a-5 antibody recognized a 50-kDa protein in 50% of the fetal adrenals. Anti-Cyp1a-2 antibody reacted with a single protein (55 kDa) in adult liver. The anti-Cyp2b-10 antibody did not detect proteins in any of the tissues. No proteins were detected in fetal kidneys. There was no coumarin 7-hydroxylase activity (COH) in fetal liver or adrenals. The 7-ethoxycoumarin O-deethylase (ECOD) activities were slightly higher in fetal adrenals (mean 6.1 pmol/mg protein/min) vs livers. The fetal adrenal ECOD activity was not inhibited by the anti-Cyp2a-5 antibody. Aryl hydrocarbon hydroxylase (AHH) activities in fetal livers were about 5% of those in adult livers. AHH activity in fetal liver was not inhibited by the anti-Cyp2c antibody. Testosterone 6 beta-hydroxylase activity was much lower in fetal liver than in adult liver (about 20 and 1700 pmol/mg protein/min, respectively). No immunoinhibition occurred in fetal adrenal progesterone hydroxylation, hepatic benzphetamine N-demethylation and hepatic ethylmorphine N-demethylation. These data suggest that members of the P450 subfamilies 1A, 2A and 2B are expressed at a very low level in fetal liver, and that fetal liver may contain members of the 2C subfamily.
针对代表Cyp1a、Cyp2a、Cyp2b和Cyp2c亚家族的纯化小鼠肝脏细胞色素P450产生的四种多克隆抗体，用于检测其在成人和胎儿组织中的相关形式。在免疫印迹分析中，抗Cyp2c抗体在成人肝脏中检测到两到三种蛋白质，在所研究的18个胎儿肝脏中的70%检测到一到三种蛋白质。抗Cyp2a-5抗体在50%的胎儿肾上腺中识别出一种50 kDa的蛋白质。抗Cyp1a-2抗体与成人肝脏中的一种单一蛋白质（55 kDa）发生反应。抗Cyp2b-10抗体在任何组织中均未检测到蛋白质。在胎儿肾脏中未检测到蛋白质。胎儿肝脏或肾上腺中没有香豆素7-羟化酶活性（COH）。胎儿肾上腺中的7-乙氧基香豆素O-脱乙基酶（ECOD）活性略高于肝脏（平均6.1 pmol/mg蛋白质/分钟）。胎儿肾上腺ECOD活性不受抗Cyp2a-5抗体的抑制。胎儿肝脏中的芳烃羟化酶（AHH）活性约为成人肝脏的5%。胎儿肝脏中的AHH活性不受抗Cyp2c抗体的抑制。胎儿肝脏中的睾酮6β-羟化酶活性远低于成人肝脏（分别约为20和1700 pmol/mg蛋白质/分钟）。胎儿肾上腺孕酮羟化、肝脏苄非他明N-脱甲基和肝脏乙基吗啡N-脱甲基均未发生免疫抑制。这些数据表明，P450亚家族1A、2A和2B的成员在胎儿肝脏中的表达水平非常低，并且胎儿肝脏可能含有2C亚家族的成员。
