A Simple, Sensitive, and Generalizable Plate Assay for Screening PARP Inhibitors

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1813; p. 245
• Main Authors: Kirby, Ilsa T, Morgan, Rory K, Cohen, Michael S
• Format: Journal Article
• Language: English
• Published: United States 2018
• Subjects: ADP Ribose Transferases - antagonists & inhibitors; ADP Ribose Transferases - chemistry; High-Throughput Screening Assays - methods; Humans; NAD - chemistry; Poly(ADP-ribose) Polymerase Inhibitors - chemistry; Poly(ADP-ribose) Polymerase Inhibitors - pharmacology; Poly(ADP-ribose) Polymerases - chemistry; Protein Processing, Post-Translational - drug effects; PARPs; Screen; ARTDs; 6-a-NAD; Click chemistry; Small-molecule inhibitor; ADP-ribosylation
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-8588-3_17

Poly-ADP-ribose polymerases (also known as ADP-ribosyltransferases or ARTDs) are a family of 17 enzymes in humans that catalyze the reversible posttranslational modification known as ADP-ribosylation. PARPs are implicated in diverse cellular processes, from DNA repair to the unfolded protein response. Small-molecule inhibitors of PARPs have improved our understanding of PARP-mediated biology and, in some cases, have emerged as promising treatments for cancers and other human diseases. However these advancements are hindered, in part, by a poor understanding of inhibitor selectivity across the PARP family. Here, we describe a simple, sensitive, and generalizable plate assay to test the potency and selectivity of small molecules against several PARP enzymes in vitro. In principle, this assay can be extended to all active PARPs, providing a convenient and direct comparison of inhibitors across the entire PARP enzyme family.