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  • Title: PARP-1 Interaction with and Activation by Histones and Nucleosomes.
    Author: Thomas C, Kotova E, Tulin AV.
    Journal: Methods Mol Biol; 2017; 1608():255-267. PubMed ID: 28695515.
    Abstract:
    Poly(ADP-ribose) Polymerase 1 (PARP-1) is an abundant chromatin associated protein, typical for most eukaryotic nuclei. The localization of PARP-1 in chromatin and its enzymatic activation involves multiple interactions of PARP-1 with nucleosomal histones, other proteins, and DNA. We report a set of methods designed to reconstitute PARP-1 regulation in vitro. These methods involve the expression of PARP-1 and PARP-1-regulating proteins using bacterial and eukaryotic systems, purification of these proteins using chromatography, testing of individual interactions in vitro, assembly of active complexes, and reconstitution of PARP-1 regulating reactions in vitro.
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