Development and validation of high-throughput screening assays for poly(ADP-ribose) polymerase-2 inhibitors
| Autor: | Jing Jin, Zhixiang Zhu, Xiaoguang Chen, Nina Xue, Xiuyun Song |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
DNA repair
High-throughput screening Drug Evaluation Preclinical Biophysics Enzyme-Linked Immunosorbent Assay Poly(ADP-ribose) Polymerase Inhibitors Biology Biochemistry chemistry.chemical_compound PARP1 Biotin High-Throughput Screening Assays medicine Humans Biotinylation Enzyme Inhibitors Molecular Biology Enzyme Assays Cancer Cell Biology NAD medicine.disease chemistry NAD+ kinase Poly(ADP-ribose) Polymerases Poly [ADP-Ribose] Polymerase 2 HeLa Cells |
| Zdroj: | Analytical Biochemistry. 449:188-194 |
| ISSN: | 0003-2697 |
| Popis: | Poly(ADP-ribose) polymerase-1 and -2 (PARP1/2) are two key facilitators of DNA repair and are implicated in the pathogenesis of cancers and several chronic diseases. Inhibitors of PARP1/2 have shown powerful therapeutic effects in the treatment of cancer, cerebral ischemia, and inflammation. In addition, evidence from several studies suggests unique functions for PARP2 in genome surveillance, spermatogenesis, adipogenesis, and T cell development, and PARP2-specific inhibitors might have many other applications. To acquire PARP1/2 inhibitors, many high-throughput screening (HTS) assays for PARP1 inhibitors have been developed. However, detailed screening assays for PARP2 inhibitors have not been reported. Herein, three HTS assays for PARP2 inhibitors were developed and validated with reference inhibitors in each case. The results suggest that the HTS assays for PARP2 inhibitors using chemical quantification of NAD + , biotin-based quantification of PAR, and ELISA quantification of PAR are sensitive, robust, and cost effective. |
| Databáze: | OpenAIRE |
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