An assay to measure poly(ADP ribose) glycohydrolase (PARG) activity in cells
| Autor: | Kerry Shea, Stephen T. Durant, Louise A. Griffiths, Emma E. Fairweather, Kay Eckersley, Dominic I. James, Ian D. Waddell, Donald J. Ogilvie, Paul P. Kelly, Mark J. O'Connor, Nicola Hamilton |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
DNA damage DNA repair Poly ADP ribose polymerase Biology DNA damage response olaparib General Biochemistry Genetics and Molecular Biology PARP Olaparib 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cell Signaling Ribose PARG General Pharmacology Toxicology and Pharmaceutics Poly(ADP-ribose) glycohydrolase Base excision repair Genetics Manchester Cancer Research Centre General Immunology and Microbiology ResearchInstitutes_Networks_Beacons/mcrc Structure: Replication & Repair Articles General Medicine Method Article MMS 030104 developmental biology chemistry Biochemistry ADP ribosylation 030220 oncology & carcinogenesis Medical Genetics |
| Zdroj: | James, D I, Durant, S, Eckersley, K, Fairweather, E, Griffiths, L A, Hamilton, N, Kelly, P, O'Connor, M, Shea, K, Waddell, I D & Ogilvie, D J 2016, ' An assay to measure poly(ADP ribose) glycohydrolase (PARG) activity in cells ', F1000Research, vol. 5, no. 736, pp. 736 . https://doi.org/10.12688/f1000research.8463.2 F1000Research |
| ISSN: | 2046-1402 |
| DOI: | 10.12688/f1000research.8463.2 |
| Popis: | After a DNA damage signal multiple polymers of ADP ribose attached to poly(ADP) ribose (PAR) polymerases (PARPs) are broken down by the enzyme poly(ADP) ribose glycohydrolase (PARG). Inhibition of PARG leads to a failure of DNA repair and small molecule inhibition of PARG has been a goal for many years. To determine whether biochemical inhibitors of PARG are active in cells we have designed an immunofluorescence assay to detect nuclear PAR after DNA damage. This 384-well assay is suitable for medium throughput high-content screening and can detect cell-permeable inhibitors of PARG from nM to µM potency. In addition, the assay has been shown to work in murine cells and in a variety of human cancer cells. Furthermore, the assay is suitable for detecting the DNA damage response induced by treatment with temozolomide and methylmethane sulfonate (MMS). Lastly, the assay has been shown to be robust over a period of several years. |
| Databáze: | OpenAIRE |
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