Stochastic Protein Alkylation by Antimalarial Peroxides

• Published in: ACS infectious diseases 2019-12, Vol.5 (12), p.2067-2075
• Main Authors: Jourdan, Joëlle, Walz, Annabelle, Matile, Hugues, Schmidt, Alexander, Wu, Jianbo, Wang, Xiaofang, Dong, Yuxiang, Vennerstrom, Jonathan L, Schmidt, Remo S, Wittlin, Sergio, Mäser, Pascal
• Format: Article
• Language: English
• ISSN: 2373-8227; 2373-8227
• DOI: 10.1021/acsinfecdis.9b00264

Antimalarial peroxides such as the phytochemical artemisinin or the synthetic ozonides arterolane and artefenomel undergo reductive cleavage of the pharmacophoric peroxide bond by ferrous heme, released by parasite hemoglobin digestion. The generated carbon-centered radicals alkylate heme in an intramolecular reaction and proteins in an intermolecular reaction. Here, we determine the proteinaceous alkylation signatures of artemisinin and synthetic ozonides in Plasmodium falciparum using alkyne click chemistry probes to identify target proteins by affinity purification and mass spectrometry-based proteomics. Using stringent controls and purification procedures, we identified 25 P. falciparum proteins that were alkylated by the antimalarial peroxides in a peroxide-dependent manner, but the alkylation patterns were more random than we had anticipated. Moreover, there was little overlap in the alkylation signatures identified in this work and those disclosed in previous studies. Our findings suggest that alkylation of parasite proteins by antimalarial peroxides is likely to be a nonspecific, stochastic process.