title: Interaction of polycyclic aromatic hydrocarbons with human cytochrome P450 1B1 in inhibiting catalytic activity.
abstract: Eleven polycyclic aromatic hydrocarbons (PAHs) and 14 acetylenic PAHs and biphenyls were used to analyze interactions with cytochrome P450 (P450) 1B1 in inhibiting catalytic activity, using 7-ethoxyresorufin O-deethylation (EROD) as a model reaction. Most of the chemicals examined were direct inhibitors of P450 1B1 except for 4-(1-propynyl)biphenyl, a mechanism-based inhibitor. In the case of direct inhibition of EROD activity {15 of 24 chemicals, e.g., benzo[a]pyrene, 1-(1-propynyl)pyrene, and 3-(1-propynyl)phenanthrene}, restoration of the EROD activity occurred with increasing incubation time, and kinetic analysis showed that EROD K(m) values were higher with these inhibitors at initial stages of incubation but became lower with increasing incubation time. With the other nine chemicals, the K(m) values for P450 1B1-mediated EROD increased during the incubations. Acetylenic inhibitors, but not the 11 PAHs, induced reverse type I spectral changes with P450 1B1, and the low dissociation constants (K(s)) suggested a role for such interaction in the inhibition of catalytic activity. Studies of quenching of P450 1B1-derived fluorescence with inhibitors demonstrated that acetylenic inhibitors and PAHs interacted rapidly with P450 1B1, with K(d) values < 10 microM. However, studies of quenching of inhibitor-derived fluorescence with P450 1B1 showed these interactions to be different, that is, B[a]P interacted with P450 1B1 more slowly. Molecular docking of P450 1B1, based on P450 1A2 crystal structure, suggested that there are clear differences in the interaction of PAH inhibitors with P450 1B1 and 1A2 and that these differences may explain why PAH inhibitors inhibit P450 1 enzymes by different mechanisms. The results suggest that P450 1B1 interacts with synthetic polycyclic aromatic acetylenes and PAHs in different ways, depending on the chemicals, and that these differences in interactions may explain how these chemicals inhibit P450 activities by different mechanisms.
authors: Laboratory of Cellular and Molecular Biology, Department of Veterinary Science, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan. t.shimada@vet.osakafu-u.ac.jp
authors: Tsutomu
authors: T
authors: Shimada
authors: Norie
authors: N
authors: Murajama
authors: Katsuhiro
authors: K
authors: Tanaka
authors: Shigeo
authors: S
authors: Takenaka
authors: Yoshio
authors: Y
authors: Imai
authors: Nancy E
authors: NE
authors: Hopkins
authors: Maryam K
authors: MK
authors: Foroozesh
authors: Willam L
authors: WL
authors: Alworth
authors: Hiroshi
authors: H
authors: Yamazaki
authors: F Peter
authors: FP
authors: Guengerich
authors: Masayuki
authors: M
authors: Komori
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D003198:Computer Simulation; D019363:Cytochrome P-450 CYP1A1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D065633:Cytochrome P-450 CYP1B1; D004791:Enzyme Inhibitors / Q000633:toxicity*; D006801:Humans; D008958:Models, Molecular; D011084:Polycyclic Aromatic Hydrocarbons / Q000633:toxicity*; D011487:Protein Conformation
chemical_list: D004791:Enzyme Inhibitors; D011084:Polycyclic Aromatic Hydrocarbons; D001189:Aryl Hydrocarbon Hydroxylases; C585496:CYP1B1 protein, human; D019363:Cytochrome P-450 CYP1A1; D065633:Cytochrome P-450 CYP1B1