An in vitro Förster resonance energy transfer-based high-throughput screening assay identifies inhibitors of SUMOylation E2 Ubc9

Yu-zhe Wang1,2, Xiao Liu3, George Way4, Vipul Madarha4, Qing-tong Zhou5, De-hua Yang1, Jia-yu Liao4, Ming-wei Wang1,2,3
1 The National Center for Drug Screening and the CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences (CAS), Shanghai 201203, China
2 University of Chinese Academy of Sciences, Beijing 100049, China
3 School of Pharmacy, Shanghai Medical College, Fudan University, Shanghai 200032, China
4 Department of Bioengineering, Bourns College of Engineering, University of California at Riverside, Riverside, CA 92521, USA
5 iHuman Institute, ShanghaiTech University, Shanghai 201210, China
Correspondence to: De-hua Yang:, Jia-yu Liao:, Ming-wei Wang:,
DOI: 10.1038/s41401-020-0405-7
Received: 20 December 2019
Accepted: 20 March 2020
Advance online: 27 April 2020


SUMOylation is one of the posttranslational modifications that mediate cellular activities such as transcription, DNA repair, and signal transduction and is involved in the cell cycle. However, only a limited number of small molecule inhibitors have been identified to study its role in cellular processes. Here, we report a Förster resonance energy transfer (FRET) high-throughput screening assay based on the interaction between E2 Ubc9 and E3 PIAS1. Of the 3200 compounds screened, 34 (1.1%) showed higher than 50% inhibition and 4 displayed dose–response inhibitory effects. By combining this method with a label-free surface plasmon resonance (SPR) assay, false positives were excluded leading to discovering WNN0605-F008 and WNN1062-D002 that bound to Ubc9 with KD values of 1.93 ± 0.62 and 5.24 ± 3.73 μM, respectively. We examined the effect of the two compounds on SUMO2-mediated SUMOylation of RanGAP1, only WNN0605-F008 significantly inhibited RanGAP1 SUMOylation, whereas WNN1062-D002 did not show any inhibition. These compounds, with novel chemical scaffolds, may serve as the initial material for developing new SUMOylation inhibitors.
Keywords: SUMOylation inhibitor; WNN0605-F008; high-throughput screening; Ubc9; PIAS1; Förster resonance energy transfer; surface plasmon resonance

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