title: The alpha-glycosidic bonds of poly(ADP-ribose) are acid-labile.
abstract: The poly(ADP-ribosyl)ation system of higher eukaryotes produces multiple ADP-ribose polymers of distinct sizes which exhibit different binding affinities for histones. Although precipitation with trichloroacetic acid (TCA) is the standard procedure for isolation of poly(ADP-ribose) from biological material, we show here that poly(ADP-ribose) is not stable under acidic conditions. Storage of poly(ADP-ribose) as TCA pellets results in acid hydrolysis of polymers, the extent of which is dependent on storage time and temperature. The alpha-glycosidic, inter-residue bonds are the preferred sites of attack, thus reducing polymer sizes by integral numbers of ADP-ribose to yield artefactually more and smaller polymers than originally present. Therefore, poly(ADP-ribosyl)ation studies involving TCA precipitation, histone extraction with acids, or acidic incubations of ADP-ribose polymers must account for the impact of acids on resulting polymer populations.
authors.affiliation: University of Zürich, Institute of Pharmacology and Biochemistry, Switzerland.
authors.forename: P L
authors.initials: PL
authors.lastname: Panzeter
authors.forename: B
authors.initials: B
authors.lastname: Zweifel
authors.forename: F R
authors.initials: FR
authors.lastname: Althaus
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D006027:Glycosides; D006868:Hydrolysis; D008970:Molecular Weight; D010761:Phosphorus Radioisotopes; D011064:Poly Adenosine Diphosphate Ribose / Q000737:chemistry* / Q000302:isolation & purification; D014238:Trichloroacetic Acid
chemical_list: D006027:Glycosides; D010761:Phosphorus Radioisotopes; D011064:Poly Adenosine Diphosphate Ribose; D014238:Trichloroacetic Acid