Development of immunological methods for the study of polycyclic aromatic hydrocarbon-DNA interactions

Thesis/Dissertation · Fri Jan 01 04:00:00 EST 1993

OSTI ID:7284352

Butch, E R

The covalent interaction of polycyclic aromatic hydrocarbon (PAH) metabolites with DNA is an important event in initiation of carcinogenesis. The development of methods to detect and analyze these DNA interactions at the low levels that occur in vivo will facilitate investigations of the mechanisms of mutagenesis and transformation by these carcinogens. Benzo[c]phenanthrene (B[c]Ph) is a polycyclic aromatic hydrocarbon with a hindered bay region': a property common to many of the most highly carcinogenic PAH. Both diastereomers of B[c]Ph-3,4-diol-1,2-epoxide (B[c]PhDE) are stable in aqueous solution, react extensively with deoxyadenosine and deoxyguanosine residues in DNA, and are potent tumor initiators in mouse-skin. B[c]PhDE-modified DNA was used to develop immunological methods for the characterization of the binding of diol epoxides of PAH with hindered bay regions'. Polyclonal antibodies were prepared in New Zealand White rabbits against DNA modified by both diastereomers of B[c]PhDE. The B[c]PhDE-DNA antisera were characterized by competitive ELISA techniques and each had distinct properties. Antiserium prepared against DNA modified with the isomer of B[c]PhDE with the 4-hydroxyl and the epoxide syn (B[c]PhDE-1) recognized B[c]PhDE-1-dA adducts and, 3-fold less efficiently, DNA modified with 7,12-dimethylbenz[a]anthracene-3,4-diol-1,2-epoxide or the isomer of B[c]PhDe with the 4-hydroxyl and epoxide anti (B[c]PhDE-2-DNA). B[c]PhDE-1-poly[dA-dT][center dot]poly[dA-dT] competed 25-fold more effectively than B[c]PhDE-1-poly[dG-dC][center dot]poly[dG-dC]. Benzo[a]pyrene-7,8-diol-9,10-epoxide-2-DNA, which contains mainly dG adducts, was not an effective competitor for binding to this antiserum even at the highest doses tested. In contrast, antisera prepared against B[c]PhDE-2-DNA is stereoselective for DNA modified by one particular stereoisomer, ([minus])-B[c]PhDE-2-DNA, and does not bind efficiently to DNA modified by any of the other PAHDE.

• Research Organization: Purdue Univ., Lafayette, IN (United States)
• OSTI ID: 7284352
• Country of Publication: United States
• Language: English

Related Subjects

560300* -- Chemicals Metabolism & Toxicology
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ADDUCTS
AROMATICS
BIOASSAY
CARCINOGENESIS
CHEMICAL REACTIONS
DETECTION
DNA ADDUCTS
HYDROCARBONS
IMMUNOASSAY
ORGANIC COMPOUNDS
PATHOGENESIS
POLYCYCLIC AROMATIC HYDROCARBONS