title: Characterization of the Artemisinin Binding Site for Translationally Controlled Tumor Protein (TCTP) by Bioorthogonal Click Chemistry.
abstract: Despite the fact that multiple artemisinin-alkylated proteins in Plasmodium falciparum have been identified in recent studies, the alkylation mechanism and accurate binding site of artemisinin-protein interaction have remained elusive. Here, we report the chemical-probe-based enrichment of the artemisinin-binding peptide and characterization of the artemisinin-binding site of P. falciparum translationally controlled tumor protein (TCTP). A peptide fragment within the N-terminal region of TCTP was enriched and found to be alkylated by an artemisinin-derived probe. MS2 fragments showed that artemisinin could alkylate multiple amino acids from Phe12 to Tyr22 of TCTP, which was supported by labeling experiments upon site-directed mutagenesis and computational modeling studies. Taken together, the "capture-and-release" strategy affords consolidated advantages previously unavailable in artemisinin-protein binding site studies, and our results deepened the understanding of the mechanism of protein alkylation via heme-activated artemisinin.
authors.affiliation: 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.|University of Chinese Academy of Sciences , Beijing 100039, China.
authors.forename: Weichao
authors.identifier: http://orcid.org/0000-0001-6875-7315
authors.initials: W
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Yiqing
authors.identifier: http://orcid.org/0000-0002-6391-3259
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School , Shenzhen 518055, China.
authors.forename: Guanghui
authors.initials: G
authors.lastname: Tang
authors.affiliation: 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.|University of Chinese Academy of Sciences , Beijing 100039, China.
authors.forename: Youli
authors.identifier: http://orcid.org/0000-0002-4803-3333
authors.initials: Y
authors.lastname: Xiao
mesh_terms: D000478:Alkylation; D037621:Artemisinins / Q000378:metabolism*; D001665:Binding Sites; D057930:Click Chemistry / Q000379:methods*; D006418:Heme / Q000737:chemistry; D013058:Mass Spectrometry; D062105:Molecular Docking Simulation; D016297:Mutagenesis, Site-Directed; D010963:Plasmodium falciparum / Q000737:chemistry*; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results
chemical_list: D037621:Artemisinins; D015800:Protozoan Proteins; D006418:Heme; C031327:artemisinin