Metabolism of N-substituted 7-methoxy-4-(aminomethyl) -coumarins by cytochrome P450 2D6 mutants and the indication of additional substrate interaction points.
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.
Keizers
P H J
PH
Department of Pharmacochemistry, Division of Molecular Toxicology, Leiden Amsterdam Center for Drug Research (LACDR), Vrije Universiteit, Amsterdam, The Netherlands.
Van Dijk
B R
BR
De Graaf
C
C
Van Vugt-Lussenburg
B M A
BM
Vermeulen
N P E
NP
Commandeur
J N M
JN
D000478:Alkylation; D001665:Binding Sites; D002384:Catalysis; D003374:Coumarins / Q000737:chemistry / Q000378:metabolism*; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008745:Methylation; D008958:Models, Molecular; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D013379:Substrate Specificity
C120605:7-methoxy-4-(aminomethyl)coumarin; D003374:Coumarins; D050505:Mutant Proteins; D019389:Cytochrome P-450 CYP2D6