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.

中文翻译：

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抗疟过氧化物的随机蛋白质烷基化。

抗疟过氧化物如植物化学青蒿素或合成臭氧化物arterolane和artefenomel通过亚铁血红素进行药效过氧化物键的还原裂解，通过寄生虫血红蛋白消化释放。生成的以碳为中心的自由基在分子内反应中烷基化血红素，在分子间反应中烷基化蛋白质。在这里，我们使用炔烃点击化学探针确定了恶性疟原虫中青蒿素和合成臭氧化物的蛋白质烷基化特征，通过亲和纯化和基于质谱的蛋白质组学来鉴定目标蛋白质。我们使用严格的控制和纯化程序，鉴定了 25 P. falciparum以过氧化物依赖性方式被抗疟过氧化物烷基化的蛋白质，但烷基化模式比我们预期的更随机。此外，在这项工作中确定的烷基化特征与之前研究中公开的那些特征几乎没有重叠。我们的研究结果表明，抗疟过氧化物对寄生虫蛋白质的烷基化可能是一个非特异性的随机过程。