Active Poly ADP Ribose Glycohydrolase (PARG)

Poly ADP Ribose Glycohydrolase (PARG) is a primary hydrolase involved in the degradation of poly(ADP-ribose) (PAR). It possesses both endo-glycohydrolase and exo-glycohydrolase activity, with a preference for the latter by binding to the two most distal ADP-ribose residues within the PAR chain. These enzymatic actions produce either free PAR or mono ADP-ribose moieties, respectively. The liberated mono ADP-ribose is further metabolized into AMP and ribose 5' phosphate by enzymes such as the NUDIX family. PARG thus plays a crucial role in regulating PAR levels, which are involved in various cellular processes including DNA damage response, chromatin maintenance, and DNA replication.Besides,Activating Transcription Factor 6 (ATF6) has been identified as an interactor of PARG, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human PARG and recombinant human ATF6. Briefly, biotin-linked PARG were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to ATF6-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of PARG and ATF6 was shown in Figure 1, the EC50 for this effect is 0.28ug/mL.