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

Gao, Peng1; Liu, Yan-Qing1; Xiao, Wei2; Xia, Fei1; Chen, Jia-Yun1; Gu, Li-Wei1; Yang, Fan3; Zheng, Liu-Hai3; Zhang, Jun-Zhe1; Zhang, Qian1; Li, Zhi-Jie3; Meng, Yu-Qing1; Zhu, Yong-Ping1; Tang, Huan1; Shi, Qiao-Li1; Guo, Qiu-Yan1; Zhang, Ying1; Xu, Cheng-Chao1; Dai, Ling-Yun3,4; Wang, Ji-Gang1,3,5

2022-06-14

10.1186/s40779-022-00390-3

Military Medical Research   Impact Factor & Quartile

2095-7467

2054-9369

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 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. 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.

Chloroquine Antimalaria Activity-based protein profiling (ABPP) Cellular thermal shift assay (CETSA) Quantitative proteomics

SCI

English

Corresponding

National Key Research and Development Program of China[2020YFA0908000] ; Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine[ZYYCXTD-C-202002] ; National Natural Science Foundation of China[82074098,82003814] ; CACMS Innovation Fund[CI2021A05101] ; Fundamental Research Funds for the Central public welfare research institutes["ZZ14-YQ-050","ZZ14-YQ-051","ZZ14-ND-010","ZZ15-ND-10","ZZ14-FL-002"]

General & Internal Medicine

Medicine, General & Internal

WOS:000810673800001

BMC

Web of Science

1.China Acad Chinese Med Sci, Artemisinin Res Ctr, Inst Chinese Mat Med, Beijing 100700, Peoples R China
2.Southern Med Univ, Dept Tradit Chinese Med, Guangzhou 510515, Peoples R China
3.Southern Univ Sci & Technol, Jinan Univ, Affiliated Hosp 1, Shenzhen Peoples Hosp,Dept Geriatr,Clin Med Coll, Shenzhen 518020, Guangdong, Peoples R China
4.ASTAR, Inst Mol & Cell Biol, Singapore 138673, Singapore
5.Natl Univ Singapore, Dept Biol Sci, Singapore 117543, Singapore

Gao, Peng,Liu, Yan-Qing,Xiao, Wei,et al. Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA[J]. Military Medical Research,2022,9(1).

Gao, Peng.,Liu, Yan-Qing.,Xiao, Wei.,Xia, Fei.,Chen, Jia-Yun.,...&Wang, Ji-Gang.(2022).Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA.Military Medical Research,9(1).

Gao, Peng,et al."Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA".Military Medical Research 9.1(2022).