Competitive profiling of celastrol targets in human cervical cancer HeLa cells via quantitative chemical proteomics.
Celastrol, isolated from the traditional Chinese medicinal herb Tripterygium wilfordii Hook. f. (Thunder God's Vine), has been used to treat cancer, chronic inflammatory, autoimmune and other human diseases. However, to date, the protein targets and the mechanism of action of celastrol have remained elusive. In this study, we find that celastrol can react with protein thiols in a unique covalent and reversible manner, while protein denaturing disrupts the interaction. Through a competitive chemoproteomics approach utilizing a cysteine-targeting activity-based probe, we report the proteome-wide quantitative profiling of cellular targets of celastrol in human cervical cancer HeLa cells. Representative targets are further validated via in vitro binding experiments and/or enzymatic activity assays. Bioinformatics analysis results suggest that celastrol exerts its numerous therapeutic effects through interaction with promiscuous proteins involved in various biological processes and cellular pathways.
CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. ylxiao@sibs.ac.cn.
Yiqing
http://orcid.org/0000-0002-6391-3259
Y
Zhou
CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. ylxiao@sibs.ac.cn and University of Chinese Academy of Sciences, Beijing 100039, China.
Weichao
W
Li
CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. ylxiao@sibs.ac.cn and Department of Chemistry, East China University of Science and Technology, Shanghai 200001, China.
Mingli
M
Wang
University of Chinese Academy of Sciences, Beijing 100039, China and Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Xixi
X
Zhang
Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Haibing
H
Zhang
Department of Chemistry, East China University of Science and Technology, Shanghai 200001, China.
Xiaofeng
X
Tong
CAS Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. ylxiao@sibs.ac.cn and University of Chinese Academy of Sciences, Beijing 100039, China.
Youli
Y
Xiao
D015415:Biomarkers; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D019295:Computational Biology / Q000379:methods; D005260:Female; D006367:HeLa Cells; D006801:Humans; D053978:Pentacyclic Triterpenes; D011485:Protein Binding; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D020543:Proteome; D040901:Proteomics / Q000379:methods; D014315:Triterpenes / Q000494:pharmacology*; D002583:Uterine Cervical Neoplasms / Q000378:metabolism*
D015415:Biomarkers; D002352:Carrier Proteins; D053978:Pentacyclic Triterpenes; D020543:Proteome; D014315:Triterpenes; C050414:celastrol