Figure 2: Enzymatic and chemical release of H-Y-E PARPs indicates that additional features of PARP catalytic domain impact to enzymatic activity. | Nature Communications

Figure 2: Enzymatic and chemical release of H-Y-E PARPs indicates that additional features of PARP catalytic domain impact to enzymatic activity.

From: Family-wide analysis of poly(ADP-ribose) polymerase activity

Figure 2

Automodified PARPs were treated with CHES or the indicated wild type and catalytically inactive ADPr hydrolytic enzyme. Signal remaining attached to protein was analysed by SDS–PAGE (top row), Coomassies of either IgG (PARPs 1, 2, 5a and 5b) or PARP (PARP3 and 4) are shown below. Released product was analysed by TLC (middle row) and sequencing gel (bottom row). Assays were repeated at least twice. PARPs 1, 2, 5a and 5b release PAR ladders upon CHES treatment and are sensitive to T. curvata PARG hydrolysis. In contrast, PARP3 and 4 release ADPr upon CHES, MacroD1 and T. curvata PARG treatment, indicative of MAR synthesis activity. See also Supplementary Fig. 4.

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