细胞色素P450 2D6突变体对N-取代的7-甲氧基-4-(氨甲基)香豆素的代谢及其他底物相互作用位点的指示

Metabolism of N-substituted 7-methoxy-4-(aminomethyl) -coumarins by cytochrome P450 2D6 mutants and the indication of additional substrate interaction points.

作者信息

Keizers P H J, Van Dijk B R, De Graaf C, Van Vugt-Lussenburg B M A, Vermeulen N P E, Commandeur J N M

机构信息

Department of Pharmacochemistry, Division of Molecular Toxicology, Leiden Amsterdam Center for Drug Research (LACDR), Vrije Universiteit, Amsterdam, The Netherlands.

出版信息

Xenobiotica. 2006 Sep;36(9):763-71. doi: 10.1080/00498250600765325.

DOI:10.1080/00498250600765325

PMID:16971342

Abstract

Previous studies have shown the critical roles residues F120 and F483 play in the oxidative metabolism of 7-methoxy-4-(aminomethyl)-coumarin (MAMC) by cytochrome P450 2D6 (CYP2D6). In the present study, a series of N-alkyl-7-methoxy-4-(aminomethyl)-coumarins (MAMC analogues) were used as substrates for the F120A and F483A mutants in order to probe the CYP2D6 active site. The F120A and F483A mutants of CYP2D6 displayed significant activity towards the MAMC analogues. Automated docking studies of the MAMC analogues in a CYP2D6 homology model suggested a distal hydrophobic active site binding cleft for the substrate N-alkyl chains, consisting of the residues L213 and V308.

摘要

先前的研究表明，残基F120和F483在细胞色素P450 2D6（CYP2D6）对7-甲氧基-4-（氨基甲基）香豆素（MAMC）的氧化代谢中发挥着关键作用。在本研究中，一系列N-烷基-7-甲氧基-4-（氨基甲基）香豆素（MAMC类似物）被用作F120A和F483A突变体的底物，以探究CYP2D6的活性位点。CYP2D6的F120A和F483A突变体对MAMC类似物表现出显著活性。在CYP2D6同源模型中对MAMC类似物进行的自动对接研究表明，底物N-烷基链存在一个由残基L213和V308组成的远端疏水活性位点结合裂隙。