title: Electrochemiluminescent arrays for cytochrome P450-activated genotoxicity screening. DNA damage from benzo[a]pyrene metabolites.
abstract: Arrays suitable for genotoxicity screening are reported that generate metabolites from cytochrome P450 enzymes (CYPs) in thin-film spots. Array spots containing DNA, various human cyt P450s, and electrochemiluminescence (ECL) generating metallopolymer [Ru(bpy)2PVP10]2+ were exposed to H2O2 to activate the enzymes. ECL from all spots was visualized simultaneously using a CCD camera. Using benzo[a]pyrene as a test substrate, enzyme activity for producing DNA damage in the arrays was found in the order CYP1B1 > CYP1A2 > CYP1A1 > CYP2E1 > myoglobin, the same as the order of their metabolic activity. Thus, these arrays estimate the relative propensity of different enzymes to produce genotoxic metabolites. This is the first demonstration of ECL arrays for high-throughput in vitro genotoxicity screening.
authors.affiliation: Department of Chemistry, 55 North Eagleville Road, University of Connecticut, Storrs, Connecticut 06269, USA.
authors.forename: Eli G
authors.initials: EG
authors.lastname: Hvastkovs
authors.forename: Minjeong
authors.initials: M
authors.lastname: So
authors.forename: Sadagopan
authors.initials: S
authors.lastname: Krishnan
authors.forename: Besnik
authors.initials: B
authors.lastname: Bajrami
authors.forename: Maricar
authors.initials: M
authors.lastname: Tarun
authors.forename: Ingela
authors.initials: I
authors.lastname: Jansson
authors.forename: John B
authors.initials: JB
authors.lastname: Schenkman
authors.forename: James F
authors.initials: JF
authors.lastname: Rusling
mesh_terms: D001564:Benzo(a)pyrene / Q000633:toxicity; D002273:Carcinogens / Q000633:toxicity*; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000187:drug effects; D004247:DNA / Q000032:analysis / Q000187:drug effects* / Q000235:genetics; D004249:DNA Damage; D004563:Electrochemistry; D004852:Epoxy Compounds / Q000032:analysis*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008163:Luminescent Measurements; D013058:Mass Spectrometry; D046228:Microarray Analysis / Q000379:methods*; D009942:Organometallic Compounds / Q000737:chemistry; D015203:Reproducibility of Results
chemical_list: D002273:Carcinogens; D004852:Epoxy Compounds; D009942:Organometallic Compounds; C492100:bis(2,2'-bipyridine)decakis(poly(vinyl pyridine))ruthenium(II); D001564:Benzo(a)pyrene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide