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Discovery of a novel OGT inhibitor through high-throughput screening based on Homogeneous Time-Resolved Fluorescence (HTRF)

文献类型：期刊论文

作者Wu, Xinyu8,9; Wang, Mingchen5,8; Cao, Yu4,8; Xu, Ying3,8; Yang, Ziqun2,7; Ding, Yiluan1,7; Lu, Jing9; Zheng, Jie2,7; Luo, Cheng4,6,7,8; Zhao, Kehao9
刊名BIOORGANIC CHEMISTRY
出版日期2023-10-01
卷号139页码:12
关键词O-GlcNAcylation O-GlcNAc transferase High-throughput screening Inhibitor
ISSN号0045-2068
DOI10.1016/j.bioorg.2023.106726
通讯作者Zhao, Kehao(kehaozhao@gmail.com) ; Chen, Shijie(shijiechen@simm.ac.cn)
英文摘要O-GlcNAcylation is a specific type of post-translational glycosylation modification, which is regulated by two enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Aberrant overexpression of OGT is associated with the development of many solid tumors. In this study, we have developed and optimized a sensitive Ho-mogeneous Time-Resolved Fluorescence (HTRF) assay then identified a novel OGT inhibitor CDDO (also called Bardoxolone) through a high-throughput screening (HTS) based on HTRF assay. Further characterization sug-gested that CDDO is an effective OGT inhibitor with an IC50 value of 6.56 +/- 1.69 mu M. CPMG-NMR analysis confirmed that CDDO is a direct binder of OGT with a binding affinity (Kd) of approximately 1.7 mu M determined by the MST analysis. Moreover, HDX-MS analysis indicated that CDDO binds to the TPR domain and N-Terminal domain of OGT, which was further confirmed by the enzymatic competition experiments as the binding of CDDO to OGT was not affected by the catalytic site binding inhibitor OSMI-4. Our docking modeling analysis further predicted the possible interactions between CDDO and OGT, providing informative molecular basis for further optimization of the inhibitor in the future. Together, our results suggested CDDO is a new inhibitor of OGT with a distinct binding pocket from the reported OGT inhibitors. Our work paved a new direction for developing OGT inhibitors driven by novel mechanisms.
WOS关键词O-GLCNAC TRANSFERASE ; BETA-N-ACETYLGLUCOSAMINE ; NUCLEOCYTOPLASMIC GLYCOSYLATION ; INSULIN-RESISTANCE ; GLCNACYLATION ; PROTEINS ; PATHWAY ; CANCER ; CELLS ; PHOSPHORYLATION
资助项目National Key R & D Program of China[2022YFC3400500] ; National Natural Science Foundation of China[21877095] ; National Natural Science Foundation of China[91853205] ; Science and Technol- ogy Commission of Shanghai Municipality[21ZR1474700] ; Youth Innovation Promotion Association of CAS[2022279] ; Natural Science Foundation of Shandong Province[ZR2022MH291]
WOS研究方向Biochemistry & Molecular Biology ; Chemistry
语种英语
WOS记录号WOS:001148470400001
出版者ACADEMIC PRESS INC ELSEVIER SCIENCE
源URL[http://119.78.100.183/handle/2S10ELR8/308918]  
专题新药研究国家重点实验室
通讯作者Zhao, Kehao; Chen, Shijie
作者单位1.Chinese Acad Sci, Shanghai Inst Mat Med, Analyt Res Ctr Organ & Biol Mol, Shanghai 201203, Peoples R China
2.Chinese Acad Sci, Shanghai Inst Mat Med, Ctr Immunol Dis, Shanghai 201203, Peoples R China
3.China Pharmaceut Univ, Nanjing 210009, Peoples R China
4.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing 210023, Jiangsu, Peoples R China
5.ShanghaiTech Univ, Sch Life Sci & Technol, 100 Haike Rd, Shanghai 201210, Peoples R China
6.Univ Chinese Acad Sci, Hangzhou Inst Adv Study, Sch Pharmaceut Sci & Technol, Hangzhou 310024, Peoples R China
7.Univ Chinese Acad Sci, 19 Yuquan Rd, Beijing 100049, Peoples R China
8.Chinese Acad Sci, Shanghai Inst Mat Med, Drug Discovery & Design Ctr, Ctr Chem Biol,State Key Lab Drug Res, Shanghai 201203, Peoples R China
9.Yantai Univ, Collaborat Innovat Ctr Adv Drug Delivery Syst & B, Minist Educ, Sch Pharm,Key Lab Mol Pharmacol & Drug Evaluat, Yantai 264005, Shandong, Peoples R China
推荐引用方式
GB/T 7714		 Wu, Xinyu,Wang, Mingchen,Cao, Yu,et al. Discovery of a novel OGT inhibitor through high-throughput screening based on Homogeneous Time-Resolved Fluorescence (HTRF)[J]. BIOORGANIC CHEMISTRY,2023,139:12.
APA Wu, Xinyu.,Wang, Mingchen.,Cao, Yu.,Xu, Ying.,Yang, Ziqun.,...&Chen, Shijie.(2023).Discovery of a novel OGT inhibitor through high-throughput screening based on Homogeneous Time-Resolved Fluorescence (HTRF).BIOORGANIC CHEMISTRY,139,12.
MLA Wu, Xinyu,et al."Discovery of a novel OGT inhibitor through high-throughput screening based on Homogeneous Time-Resolved Fluorescence (HTRF)".BIOORGANIC CHEMISTRY 139(2023):12.

入库方式： OAI收割

来源：上海药物研究所

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