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

作　　者：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 

机构地区：[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,Singa‑pore 117543,Singapore

出　　处：《Military Medical Research》2023年第1期64-77,共14页军事医学研究（英文版）

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

摘　　要：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：CHLOROQUINE ANTIMALARIA Activity-based protein profiling(ABPP) Cellular thermal shift assay(CETSA) Quantitative proteomics 

分 类 号：R531.3[医药卫生—内科学]