Human cytochromes P450 expressed in Escherichia coli: Production of specific antibodies

被引：55

作者：

Belloc, C ^{[1
]}

Baird, S ^{[1
]}

Cosme, J ^{[1
]}

Lecoeur, S ^{[1
]}

Gautier, JC ^{[1
]}

Challine, D ^{[1
]}

deWaziers, I ^{[1
]}

Flinois, JP ^{[1
]}

Beaune, PH ^{[1
]}

机构：

[1] UNIV PARIS 05,INSERM,U75,F-75730 PARIS 15,FRANCE

来源：

TOXICOLOGY | 1996年 / 106卷 / 1-3期

关键词：

human CYP; E-coli; antibody; heterologous expression; xenobiotic; metabolism;

D O I：

10.1016/0300-483X(95)03178-I

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

Cytochromes P450 (CYP) constitute a superfamily of enzymes involved in the metabolism of xenobiotics. Within the same subfamily, the isoforms present strong similarities, making them difficult to characterize and differentiate. Using heterologous expression in bacteria, five pure human CYP (1A1, 1A2, 2C9, 2E1, 3A4) were easily obtained and used as antigens to raise specific antibodies. These antibodies were characterized for their specificity and sensitivity by immunoblots; anti-CYP3A4 was immunoinhibitor. These antibodies could be used in association with other means to identify the CYPs responsible for production of a given metabolite. The use of our human recombinant CYP1A2 as antigen and the corresponding specific antibody enabled us to quantify the CYP1A2 content in 43 human livers. The average level was 69 pmol of CYP1A2/mg of microsomal proteins. Finally, these antibodies were also used to evaluate the level of heme incorporation in human microsomal CYP expressed in yeasts.

引用

页码：207 / 219

页数：13

共 46 条

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