Evaluating enzymes that generate genotoxic benzo[a]pyrene metabolites using sensor arrays.
Arrays with individually addressable, demountable electrodes coated with ultrathin DNA/enzyme films were evaluated to estimate relative rates of genotoxic bioactivation of benzo[a]pyrene (BP) for several different enzymes simultaneously. Specifically, cytochrome (cyt) P450cam, cyt P40 1A2, and myoglobin in the array were activated with H2O2 to metabolize BP to genotoxic metabolites. DNA damage by the metabolites was detected by increases in square wave voltammetric oxidation peaks using Ru(bpy)3(2+) as catalyst. Cyt P450cam and cyt P450 1A2 showed 3-fold higher activity for genotoxic bioactivation of BP than myoglobin. The ability of the arrays to generate and detect metabolite-based DNA damage simultaneously for several enzymes is a rapid and promising approach to identify and characterize enzymes involved in genotoxicity of drugs and pollutants.
Wang
Bingquan
B
Department of Chemistry, University of Connecticut, U-60, 55 North Eagleville Road, Storrs, Connecticut 06269-3060, USA.
Jansson
Ingela
I
Schenkman
John B
JB
Rusling
James F
JF
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000737:chemistry; D000818:Animals; D001564:Benzo(a)pyrene / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D019475:Camphor 5-Monooxygenase / Q000737:chemistry; D002853:Chromatography, Liquid; D019388:Cytochrome P-450 CYP1A2 / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004249:DNA Damage; D004563:Electrochemistry; D004566:Electrodes; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism; D006147:Guanine / Q000737:chemistry; D006736:Horses; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D007700:Kinetics; D013058:Mass Spectrometry; D009153:Mutagens / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D009211:Myoglobin / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry; D010084:Oxidation-Reduction; D011095:Polyethylenes / Q000737:chemistry; D016958:Pseudomonas putida / Q000201:enzymology; D000644:Quaternary Ammonium Compounds / Q000737:chemistry
D004798:Enzymes; D009153:Mutagens; D009211:Myoglobin; D009942:Organometallic Compounds; D011095:Polyethylenes; D000644:Quaternary Ammonium Compounds; C415768:tris(2,2'-bipyridyl)ruthenium(II); C041004:poly-N,N-dimethyl-N,N-diallylammonium chloride; D001564:Benzo(a)pyrene; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D006147:Guanine; D004247:DNA; D006861:Hydrogen Peroxide; D019388:Cytochrome P-450 CYP1A2; D019475:Camphor 5-Monooxygenase