Objective: To establish a method for identification of the poly(ADP-ribose) binding proteins induced by benzo (a) pyrene.
Methods: Poly (ADP-ribose) binding protein were screened by immunoprecipitation assay and further separated by high performance liquid chromatography (HPLC) and two dimensional electrophoresis, then identified by MALDI-TOF-MS/MS. The proteins sequence were identified by two methods and compared the common binding motif with literature reports.
Results: Three poly (ADP-ribose) binding proteins were identified by MALDI-TOF-MS/MS combined with immunoprecipitation assay and HPLC, and twelve poly (ADP-ribose) binding proteins were identified by MALDI-TOF-MS/MS combined with immunoprecipitation assay and two dimensional electrophoresis. Most of them have a common binding motif which was consistent with the reported.
Conclusion: Combined the immunoprecipitation assay and two dimensional electrophoresis with MALDI-TOF-MS/MS could be used to analyze the poly(ADP-ribose) binding proteins, and these proteins have a common conserved binding motif.