Further evidence for a possible role of conformation in the immunogenicity and antigenicity of the oxidative DNA lesion, 8-oxo-2'deoxyguanosine.
Damage to DNA by reactive oxygen species is acknowledged to be an important factor in a number of pathological conditions, including ageing and carcinogenesis. As a consequence, the development of methods for the sensitive detection and quantitation of oxidative DNA lesions has been of paramount importance. The oxidatively modified base product which has achieved most attention is 8-oxodeoxyguanosine (8-oxodG) and is a recognised marker of oxidative DNA damage. Although both polyclonal and monoclonal antibodies have previously been raised to 8-oxodG these have, for the most part failed to recognise this lesion within the DNA polymer. We have, through dilution cloning, produced a monoclonal antibody which appears to preferentially recognise 8-oxodG over deoxyguanosine (dG) in single-stranded oxidatively modified DNA. Such discrimination was not apparent when the DNA was double-stranded. Previous work has shown that 8-oxodG favours the syn glycosidic conformation due to steric repulsion, whereas dG assumes the anti. We present initial data that appear to support the postulate that it is these differences in conformation, in addition to structural recognition of the lesion itself, which are responsible for the discrimination, by our antibody of 8-oxodG over dG in single-stranded DNA.
Division of Chemical Pathology, Centre for Mechanisms of Human Toxicity, University of Leicester, UK. MSC5@le.ac.uk
M S
MS
Cooke
K E
KE
Herbert
P C
PC
Butler
J
J
Lunec
D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D004247:DNA / Q000276:immunology; D004249:DNA Damage; D006151:Guanosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000276:immunology; D051379:Mice; D009690:Nucleic Acid Conformation; D011817:Rabbits
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D006151:Guanosine; C046215:8-hydroxyguanosine; D004247:DNA