van Bladeren P J, Balani S K, Sayer J M, Thakker D R, Boyd D R, Ryan D E, Thomas P E, Levin W, Jerina D M
Biochem Biophys Res Commun. 1987 May 29;145(1):160-7. doi: 10.1016/0006-291x(87)91301-5.
The principal oxidative metabolites formed from benzo(c)phenanthrene (B(c)Ph) by the cytochromes P450 in liver microsomes from control and treated rats are the 3,4- and 5,6-arene oxides. A procedure is described which allows determination of the enantiomer composition and absolute configuration of these arene oxides based on HPLC separation of isomeric thiolate adducts formed with N-acetyl-L-cysteine in base. Incubation of [3H]-B(c)Ph with highly purified cytochrome P450c in a reconstituted monooxygenase system followed by trapping of the metabolically formed arene oxides as above indicated that the 3,4-oxide was predominantly the (+)-(3S,4R)-enantiomer (90%) and that the 5,6-oxide consisted mainly of the (+)-(5S,6R)-enantiomer (76%). The results are discussed in terms of their implications about the catalytic binding site of cytochrome P450c.
对照大鼠和处理大鼠肝脏微粒体中的细胞色素P450将苯并(c)菲（B(c)Ph）氧化生成的主要代谢物是3,4-和5,6-芳烃氧化物。本文描述了一种方法，该方法基于在碱性条件下与N-乙酰-L-半胱氨酸形成的异构体硫醇盐加合物的HPLC分离，来测定这些芳烃氧化物的对映体组成和绝对构型。在重组单加氧酶系统中，[3H]-B(c)Ph与高度纯化的细胞色素P450c一起温育，然后按上述方法捕获代谢生成的芳烃氧化物，结果表明3,4-氧化物主要是(+)-(3S,4R)-对映体（90%），5,6-氧化物主要由(+)-(5S,6R)-对映体（76%）组成。根据这些结果对细胞色素P450c催化结合位点的影响进行了讨论。
