| Autor: |
Yanqing Liu, Peng Gao, Qian Zhang, Yuqing Meng, Chengchao Xu, Fei Xia, Zhijie Li, Liuhai Zheng, Lingyun Dai, Jigang Wang, Jia Yun Chen, Wei Xiao, Fan Yang, Yongping Zhu, Liwei Gu, Junzhe Zhang |
| Rok vydání: |
2021 |
| Předmět: |
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| Popis: |
Background Malaria is a devastating infectious disease that disproportionally threatens hundreds of millions of people in developing countries. In the history of anti-malaria campaign, chloroquine (CQ) has played an indispensable role, however, its mechanism of action (MoA) is not fully understood. Methods We used the approach of photo-affinity labeling (PAL) in the design of a chloroquine probe and developed a combined deconvolution strategy – activity-based protein profiling (ABPP) and mass spectrometry-coupled cellular thermal shift assay (MS-CESTA) – that identified the protein targets of chloroquine in an unbiased manner in this study. Results We developed a novel photo-affinity chloroquine analog probe (CQP), which retains the antimalarial activity in the nanomole range, and identified a total of 40 proteins that specifically interacted and photo-crosslinked with CQP, which was inhibited in the presence of excess CQ. Using MS-CETSA, we identified 83 candidate interacting proteins out of a total of 3375 measured parasite proteins. Together, we identified 8 proteins as the most potential hits which were commonly identified by both methods. Conclusions We found that CQ could disrupt glycolysis and energy metabolism of malarial parasites through direct binding with some of the key enzymes, a new mechanism that is different from its known inhibitory effect of hemozoin formation. This is the first report of identifying chloroquine antimalarial targets by a parallel usage of labeled (ABPP) and label-free (MS-CETSA) methods. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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