title: Methods for purification of proteins associated with cellular poly(ADP-ribose) and PARP-specific poly(ADP-ribose).
abstract: Poly(ADP-ribose) (pADPr) is a posttranslational modification that regulates protein function through two major mechanisms: covalent modification of acceptor proteins and noncovalent binding of proteins to pADPr. pADPr is synthesized by a family of enzymes called poly(ADP-ribose) polymerases (PARPs) that are themselves major targets of pADPr modification. Here, we outline two methods for the purification of pADPr-binding proteins via pADPr purification under native conditions: purification of cellular pADPr and pADPr covalently linked to specific PARPs. Together, these methods provide complementary approaches to the identification of noncovalent pADPr-protein interactions in the cell.
authors.affiliation: Koch Institute for Integrative Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Rood
authors.forename: Anthony K L
authors.initials: AK
authors.lastname: Leung
authors.forename: Paul
authors.initials: P
authors.lastname: Chang
mesh_terms: D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D011506:Proteins; D011064:Poly Adenosine Diphosphate Ribose; D011065:Poly(ADP-ribose) Polymerases