The α-glycosidic bonds of poly(ADP-ribose) are acid-labile

• Published in: Biochemical and biophysical research communications Vol. 184; no. 1; pp. 544 - 548
• Main Authors: Panzeter, Phyllis L., Zweifel, Barbara, Althaus, Felix R.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.04.1992; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, High Pressure Liquid; Drug Stability; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Glycosides; Hydrolysis; Molecular Weight; Nucleic acids; Phosphorus Radioisotopes; Poly Adenosine Diphosphate Ribose - chemistry; Poly Adenosine Diphosphate Ribose - isolation & purification; Trichloroacetic Acid; EDTA; HPLC; TCA; Polymerization; ADP ribosylation; Acids; Chemical treatment; Reaction product
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/0006-291X(92)91229-J

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 α-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.