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Evaluating enzymes that generate genotoxic benzo[a]pyrene metabolites using sensor arrays
Bingquan Wang1, Ingela Jansson, John B Schenkman
1Department of Chemistry, University of Connecticut, U-60, 55 North Eagleville Road, Storrs, Connecticut 06269-3060, USA.
Analytical Chemistry
|March 1, 2005
Summary
This study developed a novel array for simultaneously assessing the genotoxic bioactivation of benzo[a]pyrene (BP) by multiple enzymes. Cytochrome P450 enzymes demonstrated significantly higher BP bioactivation activity than myoglobin.
Area of Science:
- Biochemistry and Environmental Toxicology
- Analytical Chemistry and Biosensing
Background:
- Benzo[a]pyrene (BP) is a polycyclic aromatic hydrocarbon pollutant requiring bioactivation to exert genotoxicity.
- Identifying enzymes responsible for BP bioactivation is crucial for understanding its environmental and health impacts.
Purpose of the Study:
- To develop and evaluate an array-based system for simultaneously measuring the genotoxic bioactivation of BP by multiple enzymes.
- To compare the relative activities of cytochrome P450cam, cytochrome P450 1A2, and myoglobin in metabolizing BP to genotoxic intermediates.
Main Methods:
- Utilized an array of individually addressable electrodes coated with DNA/enzyme films.
- Activated enzymes (cytochrome P450cam, P450 1A2, myoglobin) with H2O2 to metabolize BP.
- Detected DNA damage using square wave voltammetry with Ru(bpy)3(2+) as a catalyst.
Main Results:
- Cytochrome P450cam and P450 1A2 exhibited a 3-fold higher rate of genotoxic bioactivation of BP compared to myoglobin.
- The array successfully generated and detected DNA damage induced by BP metabolites from different enzymes simultaneously.
Conclusions:
- The developed electrode array offers a rapid and effective method for simultaneous genotoxicity assessment.
- This approach is promising for identifying and characterizing enzymes involved in the bioactivation of drugs and environmental pollutants.