Separation of poly(ADP-ribose) by high-performance liquid chromatography.

The homopolymer of ADP-ribose, poly(ADP-ribose), was synthesized in vitro by liver nuclei from NAD. The protein-poly(ADP-ribose) adducts were isolated and, after base hydrolysis or proteolysis by proteinase K, the free polymers were separated from NAD, ADP-ribose, AMP and adenosine, and quantitatively determined by reversed-phase chromatography on an Ultrasphere ODS 5-micron column. Oxidation of the polymer by sodium periodate and labeling with 3H by borotritiation maintained the polymeric structure, but its modification was detectable by the chromatographic system employed.