A phosphoproteomic approach to enrich and identify mono- and poly(ADP-ribosyl)ation sites in whole cell lysate

摘要

Poly(ADP-ribose), or PAR, is a cellular polymer implicated in DNA/RNA metabolism, cell death, and cellular stress response via its role as a post-translational modification. The founding member of the Poly(ADP-ribose) Polymerase [PARP] family, PARP1, has been tightly linked to DNA repair, leading to its role as a DNA sensitizing target for chemotherapy. The cellular response to PARP inhibition, however, is not clearly understood due to a lack of tools for studying the product of PARP enzymatic activity: mono and poly(ADP-ribose). The clinical implication can be seen in the rising number of maladies which have responded to either PARP inhibition (in humans) and/or PARP genetic loss (in mice) (see figure 1), in general these responses have not been clearly understood1. We believe mass spectrometry is a necessary tool for understanding the cellular responses to PARP inhibition, and have proposed a method for 'tagging' sites of ADP-ribosylation for identification by LC-MS/MS.