Conversion of ADP-ribose to 5'-AMP by alkaline treatment and its use for an optical quantitation of mono and poly(ADP-ribose) residues in the micromolar range

Abstract

ADP-Ribose is nearly quantitatively split to 5'-AMP by treatment with alkali at elevated temperatures. This unique behaviour, which is not shown by ADP and other adenine derivatives, was used as the basis of an optical test for the selective determination of ADPR in the presence of other adenine compounds including RNA. Poly(ADPR) could also be quantified when the polymer was degraded by poly(ADPR) glycohydrolase prior to alkaline treatment. When combined with the determination of the terminal AMP residues released by phosphodiesterase I treatment, the chain length of the polymer could be calculated. Application of the method to the quantitation of protein-bound mono(ADPR) residues in Ehrlich ascites tumor cells under different growth conditions is described.