Document Detail

Toxicity screening by electrochemical detection of DNA damage by metabolites generated in situ in ultrathin DNA-enzyme films.
MedLine Citation:
PMID:  12553846     Owner:  NLM     Status:  MEDLINE    
Rapid detection of DNA damage could serve as a basis for in vitro genotoxicity screening for new organic compounds. Ultrathin films (20-40 nm) containing myoglobin or cytochrome P450(cam) and DNA grown layer-by-layer on electrodes were activated by hydrogen peroxide, and the enzyme in the film generated metabolite styrene oxide from styrene. This styrene oxide reacted with double stranded (ds)-DNA in the same film, mimicking metabolism and DNA damage in human liver. DNA damage was detected by square wave voltammetry (SWV) by using catalytic oxidation with Ru(bpy)(3)(2+) (bpy = 2,2'-bipyridine) and by monitoring the binding of Co(bpy)(3)(3+). Damaged DNA reacts more rapidly than intact ds-DNA with Ru(bpy)(3)(3+), giving SWV peaks at approximately 1 V versus SCE that grow larger with reaction time. Co(bpy)(3)(3+) binds more strongly to intact ds-DNA, and its SWV peaks at 0.04 V decreased as DNA was damaged. Little change in SWV signals was found for incubations of DNA/enzyme films with unreactive organic controls or hydrogen peroxide. Capillary electrophoresis and HPLC-MS suggested the formation of styrene oxide adducts of DNA bases under similar reaction conditions in thin films and in solution. The catalytic SWV method was more sensitive than the Co(bpy)(3)(3+) binding assay, providing multiple measurements over a 5 min reaction time.
Liping Zhou; Jing Yang; Carmelita Estavillo; James D Stuart; John B Schenkman; James F Rusling
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  125     ISSN:  0002-7863     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2003 Feb 
Date Detail:
Created Date:  2003-01-29     Completed Date:  2003-04-01     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1431-6     Citation Subset:  IM    
Department of Chemistry, University of Connecticut, Storrs, Connecticut 06269-3060, USA.
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MeSH Terms
Biosensing Techniques / methods
Camphor 5-Monooxygenase / chemistry*
DNA Adducts / chemistry
DNA Damage*
Electrochemistry / methods
Epoxy Compounds / chemistry
Hydrogen Peroxide / chemistry
Mutagenicity Tests / methods*
Myoglobin / chemistry
Styrene / chemistry
Grant Support
Reg. No./Substance:
0/DNA Adducts; 0/Epoxy Compounds; 0/Myoglobin; 100-42-5/Styrene; 7722-84-1/Hydrogen Peroxide; 96-09-3/styrene oxide; EC 5-Monooxygenase

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