The alpha-glycosidic bonds of poly(ADP-ribose) are acid-labile.
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.
Panzeter
P L
PL
University of Zürich, Institute of Pharmacology and Biochemistry, Switzerland.
Zweifel
B
B
Althaus
F R
FR
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
D006027:Glycosides; D010761:Phosphorus Radioisotopes; D011064:Poly Adenosine Diphosphate Ribose; D014238:Trichloroacetic Acid