Impact of incorporating the 2C5 crystal structure into comparative models of cytochrome P450 2D6

被引:55
作者
Kirton, SB
Kemp, CA
Tomkinson, NP
St-Gallay, S
Sutcliffe, MJ
机构
[1] Univ Leicester, Dept Biochem, Leicester LE1 7RH, Leics, England
[2] AstraZeneca R&D Charnwood, Dept Phys & Metab Sci, Loughborough, Leics, England
[3] Univ Leicester, Dept Chem, Leicester LE1 7RH, Leics, England
关键词
molecular modeling; model validation; substrate specificity; binding determinants; principal component analysis;
D O I
10.1002/prot.10192
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cytochrome P450 2D6 (CYP2D6) metabolizes approximately one third of the drugs in current clinical use. To gain insight into its structure and function, we have produced four different sets of comparative models of 2136: one based on the structures of P450s from four different microorganisms (P450 terp, P450 eryF, P450 cam, and P450 BM3), another on the only mammalian P450 (2C5) structure available, and the other two based on alternative amino acid sequence alignments of 2D6 with all five of these structures. Principal component analysis suggests that inclusion of the 2C5 crystal structure has a profound effect on the modeling process, altering the general topology of the active site, and that the models produced differ significantly from all of the templates. The four models of 2D6 were also used in conjunction with molecular docking to produce complexes with the substrates codeine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP); this identified Glu 216 [in the F-helix; substrate recognition site (SRS) 2] as a key determinant in the binding of the basic moiety of the substrate. Our studies suggest that both Asp 301 and Glu 216 are required for metabolism of basic substrates. Furthermore, they suggest that Asp 301 (I-helix, SRS-4), a residue thought from mutagenesis studies to bind directly to the basic moiety of substrates, may play a key role in positioning the B'-C loop (SRS-1) and that the loss of activity on mutating Asp 301 may therefore be the result of an indirect effect (movement of the B'-C loop) on replacing this residue. (C) 2002 Wiley-Liss, Inc.
引用
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页码:216 / 231
页数:16
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