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Characterization of the Artemisinin Binding Site for Translationally Controlled Tumor Protein (TCTP) by Bioorthogonal Click Chemistry
journal contributionposted on 2016-11-16, 00:00 authored by Weichao Li, Yiqing Zhou, Guanghui Tang, Youli Xiao
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.
Tyr 22tumor proteinfalciparum translationallyPlasmodium falciparumN-terminal regionartemisinin-derived probepeptide fragmentMS 2 fragmentsArtemisinin Binding SiteBioorthogonal Click Chemistryartemisinin-binding peptidechemical-probe-based enrichmentTranslationally Controlled Tumor Proteinartemisinin-alkylated proteinsprotein alkylationalkylation mechanismmodeling studiesheme-activated artemisininTCTPsite-directed mutagenesisbinding sitePhe 12artemisinin-binding site