An immunochemical probe for 8,5'-cycloadenosine-5'-monophosphate and its deoxy analog in irradiated nucleic acids.
Polyclonal antisera specific for 8,5'-cycloadenosine-5'-monophosphate (8,5'-cyclo-AMP) or its deoxy analog (8,5'-cyclo-dAMP) were elicited by immunizing rabbits with a conjugate prepared by the method of Johnston et al. [Biochemistry 22, 3453-3460 (1983)]. A competitive enzyme-linked immunosorbent assay (ELISA) was developed and used to detect the formation of these products in irradiated solutions of polyadenylic acid [poly(A)] or DNA which were saturated with nitrous oxide, nitrogen, or oxygen. The 8,5'-cyclo-AMP or 8,5'-cyclo-dAMP moieties could be detected in poly(A) at 1.0 krad and in native DNA at 10 krad, respectively. The yield of 8,5'-cyclo-dAMP was found to be two to three times higher in irradiated double-stranded DNA than in single-stranded DNA. The hydroxyl radical appears to initiate 8,5'-cyclonucleotide formation in irradiated nucleic acids, as demonstrated by the inhibition of 8,5'-cyclo-AMP formation in irradiated poly(A) by dimethyl sulfoxide. However, irradiation under nitrous oxide, particularly at low doses, does not lead to the expected increases in the yield of the 8,5'-cyclonucleotide.
D000249:Adenosine Monophosphate / Q000031:analogs & derivatives* / Q000032:analysis; D003037:Cobalt Radioisotopes; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000032:analysis; D004247:DNA / Q000528:radiation effects*; D003838:Deoxyadenine Nucleotides / Q000032:analysis*; D004307:Dose-Response Relationship, Radiation; D004797:Enzyme-Linked Immunosorbent Assay; D009584:Nitrogen; D009609:Nitrous Oxide; D010100:Oxygen; D011061:Poly A / Q000528:radiation effects*; D012996:Solutions
C017847:8,5'-cycloadenosine 5'-monophosphate; D003037:Cobalt Radioisotopes; D003838:Deoxyadenine Nucleotides; D012996:Solutions; C047552:8,5'-cyclo(deoxyadenosine 5'-monophosphate); D011061:Poly A; D000249:Adenosine Monophosphate; D004247:DNA; D000242:Cyclic AMP; D009609:Nitrous Oxide; D009584:Nitrogen; D010100:Oxygen