Determination of ADP-ribose and poly(ADP-ribose) by a new radioimmunoassay.
A specific and sensitive radioimmunoassay for ADP-ribose has been developed on the basis of the selective conversion of ADP-ribose to 5'-AMP by alkaline treatment. Antibodies highly specific against 5'-AMP allowed quantification of ADP-ribose converted to 5'-AMP in the range of 1-40 pmol, and in the presence of large quantities of nucleic acids or 3'-AMP. Poly(ADP-ribose) could also be determined when degraded to ADP-ribose by poly(ADP-ribose) glycohydrolase. Determination of the chain length of purified polymer was possible by a parallel determination of ADP-ribose residues after glycohydrolase treatment and of 5'-AMP from the non-reducing end obtained by phosphodiesterase catalyzed hydrolysis. The high specificities of the alkaline conversion of ADP-ribose to 5'-AMP and of the radioimmunoassay for 5'-AMP allowed quantification of protein-bound ADP-ribose residues in crude tissue extracts as verified by comparison with chromatographically purified samples.
D000247:Adenosine Diphosphate Sugars / Q000032:analysis*; D000818:Animals; D000831:Animals, Newborn; D002286:Carcinoma, Ehrlich Tumor / Q000032:analysis; D003429:Cross Reactions; D008099:Liver / Q000032:analysis; D051379:Mice; D008970:Molecular Weight; D009702:Nucleoside Diphosphate Sugars / Q000032:analysis*; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis*; D011863:Radioimmunoassay / Q000379:methods; D051381:Rats; D012266:Ribose
D000247:Adenosine Diphosphate Sugars; D009702:Nucleoside Diphosphate Sugars; D011064:Poly Adenosine Diphosphate Ribose; D012266:Ribose