Binding of bufuralol, dextromethorphan, and 3,4-methylenedioxymethylamphetamine to wild-type and F120A mutant cytochrome P450 2D6 studied by resonance Raman spectroscopy.
Cytochrome P450 2D6 (CYP2D6) is one of the most important drug-metabolizing enzymes in humans. Resonance Raman data, reported for the first time for CYP2D6, show that the CYP2D6 heme is found to be in a six-coordinated low-spin state in the absence of substrates, and it is perturbed to different extents by bufuralol, dextromethorphan, and 3,4-methylenedioxymethylamphetamine (MDMA). Dextromethorphan and MDMA induce in CYP2D6 a significant amount of five-coordinated high-spin heme species and reduce the polarity of its heme-pocket, whereas bufuralol does not. Spectra of the F120A mutant CYP2D6 suggest that Phe120 is involved in substrate-binding of dextromethorphan and MDMA, being responsible for the spectral differences observed between these two compounds and bufuralol. These differences could be explained postulating a different substrate mobility for each compound in the CYP2D6 active site, consistently with the role previously suggested for Phe120 in binding dextromethorphan and MDMA.
Laser Centre/Analytical Chemistry and Applied Spectroscopy, Vrije Universiteit Amsterdam, The Netherlands.
Alois
A
Bonifacio
Peter H J
PH
Keizers
Jan N M
JN
Commandeur
Nico P E
NP
Vermeulen
Bruno
B
Robert
Cees
C
Gooijer
Gert
G
van der Zwan
D019943:Amino Acid Substitution; D001665:Binding Sites; D019389:Cytochrome P-450 CYP2D6 / Q000737:chemistry* / Q000235:genetics; D003915:Dextromethorphan / Q000737:chemistry* / Q000378:metabolism; D004983:Ethanolamines / Q000737:chemistry* / Q000378:metabolism; D006418:Heme / Q000737:chemistry; D006801:Humans; D009154:Mutation; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000737:chemistry* / Q000378:metabolism; D013059:Spectrum Analysis, Raman
D004983:Ethanolamines; D006418:Heme; D003915:Dextromethorphan; C010831:bufuralol; D019389:Cytochrome P-450 CYP2D6; D018817:N-Methyl-3,4-methylenedioxyamphetamine