Protein engineering approach to enhance activity assays of mono-ADP-ribosyltransferases through proximity.

ABSTRACT: Human mono-ADP-ribosylating PARP enzymes have been linked to several clinically relevant processes and many of these PARPs have been suggested as potential drug targets. Despite recent advances in the field, efforts to discover inhibitors have been hindered by the lack of tools to rapidly screen for high potency compounds and profile them against the different enzymes. We engineered mono-ART catalytic fragments to be incorporated into a cellulosome-based octavalent scaffold. Compared to the free enzymes, the scaffold-based system results in an improved activity for the tested PARPs due to improved solubility, stability and the proximity of the catalytic domains, altogether boosting their activity beyond 10-fold in the case of PARP12. This allows us to measure their activity using a homogen

SUBMITTER: Galera-Prat A 

PROVIDER: S-EPMC9574550 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature