Purification of ADP-ribosylated nuclear proteins by covalent chromatography on dihydroxyboryl polyacrylamide beads and their characterization.
- 1 March 1978
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 75 (3), 1111-1115
- https://doi.org/10.1073/pnas.75.3.1111
Abstract
Nuclear proteins modified by mono or poly ADP-ribosylation were selectively isolated and purified by covalent chromatography on a dihydroxyboryl polyacrylamide bead column that specifically interacts with cis-diol-containing compounds. From rat liver nuclei that had been incubated with NAD+, histones and some nonhistone proteins were extracted with 0.25 M HCl. Approximately 60% of the ADP-ribose incorporated into 20% trichloroacetic acid-precipitable material was recovered in this extract. The ADP-ribosylated material was then isolated from the extract by covalent chromatography on a borate gel column and further purified by carboxymethylcellulose column chromatography. As judged by electrophoretic mobilities in various gel systems and by amino acid compositions, approximately 50% of the ADP-ribose recovered in the carboxymethylcellulose fractions was associated with several nonhistone proteins with molecular weights of 2-6 x 10(4), while 35% aand 15% were associated with histones H2B and H1, respectively. Since the average chain length of the polymer bound to any of these proteins was less than two ADP-ribos-l units, the percentage distribution reflects the number of ADP-ribosylated sites rather than the chain length.This publication has 34 references indexed in Scilit:
- Adenosine Diphosphoribosylation of Certain Basic Chromosomal Proteins in Isolated Trout Testis NucleiEuropean Journal of Biochemistry, 1977
- Poly (ADP-Ribose) and ADP-Ribosylation of ProteinsAnnual Review of Biochemistry, 1977
- Dissociation and reconstitution of chromatin without appreciable degradation of the proteinsBiochemical and Biophysical Research Communications, 1975
- Inhibition of rat liver Ca2+, Mg2+-dependent endonuclease activity by nicotinamide adenine dinucleotide and poly(adenosine diphosphate ribose) synthetaseBiochemical and Biophysical Research Communications, 1974
- In vivo occurence of bound ADP‐riboseFEBS Letters, 1973
- Identification of poly(ADP-ribose) covalently bound to histone F1 in vivoBiochemical and Biophysical Research Communications, 1973
- Stimulation of DNA synthesis by adenosine diphosphoribosylation of HeLa nuclear proteins during the cell cycleBiochemical and Biophysical Research Communications, 1973
- Mode of inhibition of DNA synthesis induced by adenosine diphosphoribosylation of nuclear proteinBiochemical and Biophysical Research Communications, 1971
- Derivatization of cross-linked polyacrylamide beads. Controlled introduction of functional groups for the preparation of special-purpose, biochemical adsorbentsBiochemistry, 1969
- Isolation of pure and unaltered liver nuclei morphology and biochemical compositionExperimental Cell Research, 1956