Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA

Peng Gao; Yan-Qing Liu; Wei Xiao; Fei Xia; Jia-Yun Chen; Li-Wei Gu; Fan Yang; Liu-Hai Zheng; Jun-Zhe Zhang; Qian Zhang; Zhi-Jie Li; Yu-Qing Meng; Yong-Ping Zhu; Huan Tang; Qiao-Li Shi; Qiu-Yan Guo; Ying Zhang; Cheng-Chao Xu; Ling-Yun Dai; Ji-Gang Wang

doi:10.1186/s40779-022-00390-3

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Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA

RESEARCH | Updated：2023-03-23

- Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA
- Military Medical Research Vol. 10, Issue 1, Pages: 64-77(2023)
- Affiliations：
  
  1.Artemisinin Research Center, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700 , China
  2.Department of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515 , China
  3.Department of Geriatrics, the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology, Shenzhen People’s Hospital, Shenzhen 518020 , Guangdong, China
  4.Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), Singapore 138673 , Singapore
  5.Department of Biological Sciences, National University of Singapore, Singapore 117543 , Singapore
- Author bio：
  
  * ccxu@icmm.ac.cn;
  dai.lingyun@szhospital.com;
  jgwang@icmm.ac.cn
- Funds：
- DOI：10.1186/s40779-022-00390-3
  CLC：
- Published：2023-02
- Accepted：
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Gao P, Liu YQ, Xiao W, Xia F, Chen JY, Gu LW, et al. Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA. Mil Med Res. 2022;9(1):30.
DOI：

Gao P, Liu YQ, Xiao W, Xia F, Chen JY, Gu LW, et al. Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA. Mil Med Res. 2022;9(1):30. DOI： 10.1186/s40779-022-00390-3.

摘要

Abstract

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 principle of photo-affinity labeling and click chemistry-based functionalization in the design of a CQ probe and developed a combined deconvolution strategy of activity-based protein profiling (ABPP) and mass spectrometry-coupled cellular thermal shift assay (MS-CETSA) that identified the protein targets of CQ in an unbiased manner in this study. The interactions between CQ and these identified potential protein hits were confirmed by biophysical and enzymatic assays.

Results:

We developed a novel clickable

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 photocrosslinked 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. At the same time

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 well-known inhibitory effect of hemozoin formation. This is the first report of identifying CQ antimalarial targets by a parallel usage of labeled (ABPP) and label-free (MS-CETSA) methods.

关键词

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references

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