Surface-enhanced resonance Raman scattering of cytochrome P450-2D6 on coated silver hydrosols.
Surface-enhanced resonance Raman scattering (SERRS) from dilute solutions (down to nanomolar concentrations) of human mono-oxygenase CYP2D6 is observed using aqueous dispersions of Ag nanoparticles (hydrosol) coated with self-assembled monolayers (SAMs) of mercaptoalkanoic acids of two different lengths. From a direct comparison with its resonance Raman spectrum in solution, CYP2D6 appears to fully retain its native structure upon adsorption on coated hydrosol through electrostatic interaction, while a structural change in the active site is observed when uncoated citrate-reduced hydrosol is used. Using SERRS on these biocompatible coated hydrosols, the effects of dextromethorphan on the enzyme's active site can be observed, demonstrating that CYP2D6 ability of binding substrates is preserved. Moreover, by tuning the wavelength of the exciting laser away from the main absorption band of the heme, the vibrational bands of the SAM coating are observed and analyzed to see how the presence of the protein affects the SAM structure.
Bonifacio
Alois
A
Laser Centre/Analytical Chemistry and Applied Spectroscopy, and LACDR/Molecular Toxicology, Department of Chemistry and Pharmaceutical Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
Keizers
Peter H J
PH
Vermeulen
Nico P E
NP
Commandeur
Jan N M
JN
Gooijer
Cees
C
van der Zwan
Gert
G
D019389:Cytochrome P-450 CYP2D6 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D012834:Silver / Q000737:chemistry*; D013059:Spectrum Analysis, Raman; D013379:Substrate Specificity
D012834:Silver; D019389:Cytochrome P-450 CYP2D6