An HTS-compatible HTRF assay measuring the glycohydrolase activity of human PARG.
Authors
Stowell, Alexandra I JJames, Dominic I
Waddell, Ian D
Bennett, N
Truman, C
Hardern, I
Ogilvie, Donald J
Affiliation
Cancer Research UK Manchester Institute Drug Discovery Unit, University of Manchester, Manchester,Issue Date
2016-03-29
Metadata
Show full item recordAbstract
Poly(ADP-ribose)(PAR) polymers are transient post-translational modifications, and their formation is catalyzed by poly(ADP-ribose) polymerase (PARP) enzymes. A number of PARP inhibitors are in advanced clinical development for BRCA-mutated breast cancer, and olaparib has recently been approved for BRCA-mutant ovarian cancer; however, there has already been evidence of developed resistance mechanisms. Poly(ADP-ribose) glycohydrolase (PARG) catalyzes the hydrolysis of the endo- and exo-glycosidic bonds within the PAR polymers. As an alternative strategy, PARG is a potentially attractive therapeutic target. There is only one PARG gene, compared with 17 known PARP family members, and therefore a PARG inhibitor may have wider application with fewer compensatory mechanisms. Prior to the initiation of this project, there were no known existing cell-permeable small molecule PARG inhibitors for use as tool compounds to assess these hypotheses, and no suitable high-throughput screening (HTS)-compatible biochemical assays available to identify start points for a drug discovery project. The development of this newly-described high-throughput homogeneous time-resolved fluorescence (HTRF) assay has allowed HTS to proceed, and from this, the identification and advancement of multiple validated series of tool compounds for PARG inhibition.Citation
An HTS-compatible HTRF assay measuring the glycohydrolase activity of human PARG. 2016: Anal BiochemJournal
Analytical BiochemistryDOI
10.1016/j.ab.2016.03.016PubMed ID
27036617Type
ArticleLanguage
enISSN
1096-0309ae974a485f413a2113503eed53cd6c53
10.1016/j.ab.2016.03.016
Scopus Count
Collections
Related articles
- A nonradiometric, high-throughput assay for poly(ADP-ribose) glycohydrolase (PARG): application to inhibitor identification and evaluation.
- Authors: Putt KS, Hergenrother PJ
- Issue date: 2004 Oct 15
- Discovery and structure-activity relationships of modified salicylanilides as cell permeable inhibitors of poly(ADP-ribose) glycohydrolase (PARG).
- Authors: Steffen JD, Coyle DL, Damodaran K, Beroza P, Jacobson MK
- Issue date: 2011 Aug 11
- Discovery of novel poly(ADP-ribose) glycohydrolase inhibitors by a quantitative assay system using dot-blot with anti-poly(ADP-ribose).
- Authors: Okita N, Ashizawa D, Ohta R, Abe H, Tanuma S
- Issue date: 2010 Feb 19
- A quantitative assay reveals ligand specificity of the DNA scaffold repair protein XRCC1 and efficient disassembly of complexes of XRCC1 and the poly(ADP-ribose) polymerase 1 by poly(ADP-ribose) glycohydrolase.
- Authors: Kim IK, Stegeman RA, Brosey CA, Ellenberger T
- Issue date: 2015 Feb 6
- Inhibition of poly(ADP-ribose) polymerase-1 or poly(ADP‑ribose) glycohydrolase individually, but not in combination, leads to improved chemotherapeutic efficacy in HeLa cells.
- Authors: Feng X, Koh DW
- Issue date: 2013 Feb