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Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors
Institute of Materia Medica, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100050, China
Department of Macromolecular Science and Engineering, Case Western Reserve University, Cleveland, OH 44106, USA
* Author to whom correspondence should be addressed.
Received: 4 November 2011; in revised form: 6 December 2011 / Accepted: 13 December 2011 / Published: 20 December 2011
Abstract: A disintegrin and metalloprotease with thrombospondin type I motifs-1 (ADAMTS1) plays a crucial role in inflammatory joint diseases and its inhibitors are potential candidates for anti-arthritis drugs. For the purposes of drug discovery, we reported the development and validation of fluorescence resonance energy transfer (FRET) assay for high-throughput screening (HTS) of the ADAMTS1 inhibitors. A FRET substrate was designed for a quantitative assay of ADAMTS1 activity and enzyme kinetics studies. The assay was developed into a 50-µL, 384-well assay format for high throughput screening of ADAMTS1 inhibitors with an overall Z’ factor of 0.89. ADAMTS1 inhibitors were screened against a diverse library of 40,960 total compounds with the established HTS system. Four structurally related hits, naturally occurring compounds, kuwanon P, kuwanon X, albafuran C and mulberrofuran J, extracted from the Chinese herb Morus alba L., were identified for further investigation. The results suggest that this FRET assay is an excellent tool, not only for measurement of ADAMTS1 activity but also for discovery of novel ADAMTS1 inhibitors with HTS.
Keywords: ADAMTS1; inhibitor; high-throughput screening; fluorescence resonance energy transfer; drug discovery
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MDPI and ACS Style
Peng, J.; Gong, L.; Si, K.; Bai, X.; Du, G. Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors. Molecules 2011, 16, 10709-10721.
Peng J, Gong L, Si K, Bai X, Du G. Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors. Molecules. 2011; 16(12):10709-10721.
Peng, Jianhao; Gong, Lili; Si, Kun; Bai, Xiaoyu; Du, Guanhua. 2011. "Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors." Molecules 16, no. 12: 10709-10721.