The Development of a Biotinylated NAD+-Applied Human Poly(ADP-Ribose) Polymerase 3 (PARP3) Enzymatic Assay

  title={The Development of a Biotinylated NAD+-Applied Human Poly(ADP-Ribose) Polymerase 3 (PARP3) Enzymatic Assay},
  author={Ming Ji and Liyuan Wang and Nina Xue and Fangfang Lai and Sen Zhang and Jing Jin and Xiaoguang Chen},
  journal={SLAS Discovery},
  pages={545 - 553}
Poly(ADP-ribose) polymerase 3 (PARP3) is an important member of the PARP family and shares high structural similarities with both PARP1 and PARP2. The biological roles of PARP3 are currently under investigation; however, several key reports indicate the integral roles of PARP3 in DNA damage repair, and thus it has been investigated as a novel target in oncology. It is clear that the identification of selective PARP3 inhibitors would further advance the understanding of the biological roles of… Expand
4 Citations
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These active site probes for use in in vitro and cellular biophysical assays to characterize active site-directed inhibitors that compete for NAD+ binding are agnostic of the protein substrate for each PARP, overcoming a general lack of knowledge around the substrates for these enzymes. Expand
ADP-ribosylation of RNA and DNA: from in vitro characterization to in vivo function
An overview of the enzymes that ADP-ribosylate nucleic acids, the reversing hydrolases, and the substrates’ requirements is provided, and possible functions for nucleic acid ADp- ribosylation in mammals are discussed. Expand
Forced Self-Modification Assays as a Strategy to Screen MonoPARP Enzymes
A family-wide approach to developing robust high-throughput monoPARP assays where the enzymes are immobilized and forced to self-modify using biotinylated-NAD+, which is detected using a dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA) readout. Expand
Combination of Selective PARP3 and PARP16 Inhibitory Analogues of Latonduine A Corrects F508del-CFTR Trafficking
The structure activity relationship (SAR) study reported herein has resulted in the discovery of the modestly potent (IC50 3.1 μM) PARP3 selective inhibitor (±)-5-hydroxy-4-phenyl-2,3,4,5-tetrahydro-1H-benzo[c]azepin-1-one that shows 205-fold selectivity for PARP16 compared withPARP3 in vitro. Expand


Poly(ADP-ribose) polymerase 3 (PARP3), a newcomer in cellular response to DNA damage and mitotic progression
PARP3 is described as a newcomer in genome integrity and mitotic progression and a particular role of PARP3 in cellular response to double-strand breaks is reported, most likely in concert with PARP1. Expand
An enzymatic assay for poly(ADP-ribose) polymerase-1 (PARP-1) via the chemical quantitation of NAD(+): application to the high-throughput screening of small molecules as potential inhibitors.
A highly sensitive, inexpensive, and operationally simple assay for the rapid assessment of PARP activity that relies on the conversion of NAD+ into a highly fluorescent compound and can readily detect PARP inhibitors in a high-throughput screen using 384-well plates. Expand
PARP-3 Is a Mono-ADP-ribosylase That Activates PARP-1 in the Absence of DNA*
It is reported that PARP-3 small interfering RNA-depleted cells are not sensitive to the topoisomerase I poison camptothecin, inducing DNA single-strand breaks, and repair these lesions as efficiently as wild-type cells. Expand
Poly(ADP-ribose) polymerases in double-strand break repair: focus on PARP1, PARP2 and PARP3.
The role of the DNA-activated PARP1, PARP2 and PARP3 in cellular response to double-strand breaks (DSB) is provided and the biological significance of these properties in response to programmed DNA lesions formed during physiological processes such as antibody repertoire assembly and diversification is outlined. Expand
Development and validation of high-throughput screening assays for poly(ADP-ribose) polymerase-2 inhibitors.
Three high-throughput screening assays for PARP2 inhibitors using chemical quantification of NAD(+), biotin-basedquantification of PAR, and ELISA quantifying of PAR are sensitive, robust, and cost effective. Expand
PARP inhibitor with selectivity toward ADP-ribosyltransferase ARTD3/PARP3.
The results show that by targeting the nicotinamide binding site, selective inhibition can be achieved among the closest relatives of the validated clinical target, ADP-ribosyltransferase-1/poly(ADp-ribose) polymerase- 1. Expand
Development of a high-throughput screening-amenable assay for human poly(ADP-ribose) polymerase inhibitors.
The present study demonstrates a sensitive and reproducible methodology capable of screening human PARP inhibitors in high-throughput format and validate a nonradioactive PARP assay suitable for HTS. Expand
PARP-2 and PARP-3 are selectively activated by 5′ phosphorylated DNA breaks through an allosteric regulatory mechanism shared with PARP-1
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New insights into the molecular and cellular functions of poly(ADP-ribose) and PARPs
This work has shown that the activity of PARP family members, such as PARP1 and PARP2, is tied to cellular signalling pathways, and through poly(ADP-ribosyl)ation (PARylation) they ultimately promote changes in gene expression, RNA and protein abundance, and the location and activity of proteins that mediate signalling responses. Expand
Preclinical selection of a novel poly(ADP-ribose) polymerase inhibitor for clinical trial
A compound, AG14447, is identified as a PARP inhibitor with outstanding in vivo chemosensitization potency at tolerable doses, which is at least 10 times more potent than the initial lead,AG14361. Expand