title: [Study of interaction of cytochrome P450 2B4 with riboflavin by fluorescence spectrometry].
abstract: Fluorescence quenching of riboflavin by cytochrome P450 2B4 was used to probe the ligand--enzyme binding interaction ((lambda ex = 385 nm, lambda em = 520 nm). Riboflavin is a component of a flavoprotein NADPH dependent cytochrome P450 reductase, an essential electron carrier during cytochrome P450 catalysis. Fluorescence titration measurements revealed that cytochrome P450 2B4 and riboflavin formed a complex with an apparent Kd = 8.8 +/- 1 microM. The fluorescence intensity of riboflavin decreased upon the addition of cytochrome P450 2B4, which may be caused by the resonance excitation energy transfer from the fluorescent donor riboflavin to the cytochrome P450 2B4 heme acceptor. These data suggest that there may exist specific sites of binding of riboflavin with the protein globule of cytochrome P450 2B4.
authors.affiliation: Research Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Pogodinskaya ul. 10, Moscow, 119992 Russia.victoria@ibmh.msk.su
authors.forename: V V
authors.initials: VV
authors.lastname: Shumiantseva
authors.forename: N A
authors.initials: NA
authors.lastname: Petushkova
authors.forename: T V
authors.initials: TV
authors.lastname: Bulko
authors.forename: N F
authors.initials: NF
authors.lastname: Samenkova
authors.forename: G P
authors.initials: GP
authors.lastname: Kuznetsova
authors.forename: A I
authors.initials: AI
authors.lastname: Archakov
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D002384:Catalysis; D000072467:Cytochrome P450 Family 2; D012256:Riboflavin / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D001189:Aryl Hydrocarbon Hydroxylases; D000072467:Cytochrome P450 Family 2; C465241:cytochrome P-450 CYP2B4 (rabbit); D012256:Riboflavin