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.
D000818:Animals; D001894:Borohydrides; D002467:Cell Nucleus / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism; D008970:Molecular Weight; D009243:NAD / Q000032:analysis; D009702:Nucleoside Diphosphate Sugars / Q000302:isolation & purification*; D010084:Oxidation-Reduction; D010504:Periodic Acid; D011064:Poly Adenosine Diphosphate Ribose / Q000138:chemical synthesis / Q000302:isolation & purification*; D051381:Rats
D001894:Borohydrides; D009702:Nucleoside Diphosphate Sugars; D009243:NAD; D010504:Periodic Acid; D011064:Poly Adenosine Diphosphate Ribose; C025364:sodium borohydride