title: Purification of human PARP-1 and PARP-1 domains from Escherichia coli for structural and biochemical analysis.
abstract: A general method to express and purify full-length human poly(ADP-ribose) polymerase-1 (PARP-1), individual PARP-1 domains, and groups of PARP-1 domains from Escherichia coli cells is described. The procedure allows for robust production of highly pure PARP-1 that is free of DNA contamination and well-suited for biochemical experiments and for structural and biophysical analysis. Two biochemical assays for monitoring PARP-1 automodification activity are presented that can be used to evaluate purified PARP-1, combinations of PARP-1 domains, or PARP-1 mutants.
authors.affiliation: Department of Biochemistry, Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA.
authors.forename: Marie-France
authors.initials: MF
authors.lastname: Langelier
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Planck
authors.forename: Kristin M
authors.initials: KM
authors.lastname: Servent
authors.forename: John M
authors.initials: JM
authors.lastname: Pascal
mesh_terms: D018360:Crystallography, X-Ray; D004926:Escherichia coli; D006801:Humans; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases