机构:[1]State Key Laboratory of Chemical Biology, Center for Excellence in Molecular Synthesis, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai 200032, China.[2]University of Chinese Academy of Sciences, Beijing 101408, China.[3]Institute for Agri-Food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China.[4]School of Life Science and Technology, ShanghaiTech University, Haike Road 100, Shanghai 201210, China.[5]Shanghai Tongren Hospital, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Shanghai Jiao Tong University School of Medicine, Chongqing South Road 227, Shanghai 200025, China.[6]School of Physical Science and Technology, ShanghaiTech University, Haike Road 100, Shanghai 201210, China.
Salvianolic acid A (SAA), a major active ingredient of Salvia miltiorrhiza Bunge (Danshen), displays strong antiproliferative activity against cancer cells. However, their protein targets remain unknown. Here, we deconvoluted the protein targets of SAA using chemoproteomics and phosphoproteomics. By using alkynylated SAA as a probe, we discovered that SAA is a covalent ligand that can modify cellular proteins via its electrophilic α,β-unsaturated ester moiety. The subsequent chemoproteomics profiling revealed that 46 proteins were covalently modified by SAA, including Raptor, a subunit of mTORC1 for recruiting substrates for mTORC1. Although gene ontology enrichment analysis of these proteins suggested that SAA displays a promiscuous protein interaction, phosphoproteomics profiling revealed that the SAA modulated phosphoproteins were mainly enriched in the signaling pathways of PI3K-Akt-mTOR, which is closely related to cell growth and proliferation. This was confirmed by the biochemical assay with purified mTORC1, a Western blot assay with phospho-specific antibodies, and a cellular thermal shift assay. Our work discovered that SAA is a covalent ligand for protein modification and mTORC1 is one of its targets. Moreover, our work demonstrated that the integrative profiling of chemoproteomics and phosphoproteomics can be a powerful tool for target deconvolution for bioactive natural products.
基金:
We gratefully acknowledge Prof. Yanhui Xu at the Fudan
University for providing the reagent, National Natural Science
Foundations of China (22274165, 92053101, 21775158 to
J.K.), and the Strategic Priority Research Program of the
Chinese Academy of Sciences (Grant No. XDB20020200).
第一作者机构:[1]State Key Laboratory of Chemical Biology, Center for Excellence in Molecular Synthesis, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai 200032, China.[2]University of Chinese Academy of Sciences, Beijing 101408, China.
共同第一作者:
通讯作者:
通讯机构:[1]State Key Laboratory of Chemical Biology, Center for Excellence in Molecular Synthesis, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, 345 Lingling Road, Shanghai 200032, China.[6]School of Physical Science and Technology, ShanghaiTech University, Haike Road 100, Shanghai 201210, China.[*1]State Key Laboratory of Chemical Biology, Center for Excellence in Molecular Synthesis, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China[*2]School of Physical Science and Technology, ShanghaiTech University, Shanghai 201210, China
推荐引用方式(GB/T 7714):
Zheng Mengmeng,Zhang Yanmei,Xu Yao,et al.Chemoproteomics and Phosphoproteomics Profiling Reveals Salvianolic Acid A as a Covalent Inhibitor of mTORC1[J].JOURNAL OF PROTEOME RESEARCH.2023,22(7):2450-2459.doi:10.1021/acs.jproteome.3c00188.
APA:
Zheng Mengmeng,Zhang Yanmei,Xu Yao,Han Ying,Wu Yingli&Kang Jingwu.(2023).Chemoproteomics and Phosphoproteomics Profiling Reveals Salvianolic Acid A as a Covalent Inhibitor of mTORC1.JOURNAL OF PROTEOME RESEARCH,22,(7)
MLA:
Zheng Mengmeng,et al."Chemoproteomics and Phosphoproteomics Profiling Reveals Salvianolic Acid A as a Covalent Inhibitor of mTORC1".JOURNAL OF PROTEOME RESEARCH 22..7(2023):2450-2459