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

作　　者：Yu-zhe Wang Xiao Liu George Way Vipul Madarha Qing-tong Zhou De-hua Yang Jia-yu Liao Ming-wei Wang 

机构地区：[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]Human Institute,ShanghaiTech University,Shanghai 201210,China

出　　处：《Acta Pharmacologica Sinica》2020年第11期1497-1506,共10页中国药理学报（英文版）

基　　金：supported by the National Natural Science Foundation of China grants 81872915(MWW),81573479(DHY),81773792(DHY),and 21704064(QTZ);The National Science&Technology Major Project"Key New Drug Creation and Manufacturing Program"(2018ZX09735-001 to MWW and 2018ZX09711002-002-005 to DHY);The National Key R&D Program of China grant 2018YFA0507000(MWW);Novo Nordisk-CAS Research Fund grant NNCAS-2017-1-CC(DHY).

摘　　要：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 ide ntified to study its role in cellular processes.Here,we report a Forster res on ance 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%inhibit!on and 4 displayed dose-response inhibitory effects.By combining this method with a label-free surface plasm on 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 RanGAPI,only WNN0605-F008 significantly inhibited RanGAP!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.

关 键 词：SUMOylation inhibitor WNN0605-F008 high-throughput screening UBC9 PIAS1 Forster resonance energy transfer surface plasm on resonance 

分 类 号：R96[医药卫生—药理学]