Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA.
Identification of antimalarial targets of chloroquine by a combined deconvolution strategy of ABPP and MS-CETSA.
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DOI：
10.1186/s40779-022-00390-3
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发表时间：
2022-06-14
影响因子：
21.1
通讯作者：
Wang JG
中科院分区：
文献类型：
--
作者：
Gao P;Liu YQ;Xiao W;Xia F;Chen JY;Gu LW;Yang F;Zheng LH;Zhang JZ;Zhang Q;Li ZJ;Meng YQ;Zhu YP;Tang H;Shi QL;Guo QY;Zhang Y;Xu CC;Dai LY;Wang JG
疟疾是一种毁灭性的传染病，对发展中国家数亿人构成严重威胁。在抗疟史上，氯喹（CQ）发挥了不可或缺的作用，但其作用机制尚不完全清楚。我们使用的原理，光亲和标记和点击化学为基础的功能化的CQ探针的设计，并开发了一个联合的反卷积策略的活性为基础的蛋白质谱（ABPP）和质谱耦合细胞热位移分析（MS-CETSA），确定在本研究中的蛋白质靶点的CQ。CQ和这些鉴定的潜在蛋白质命中之间的相互作用通过生物物理和酶测定来证实。我们开发了一种新型的可点击的光亲和性氯喹类似物探针（CQP），其在纳摩尔范围内保留抗疟活性，并鉴定了总共40种与CQP特异性相互作用和光交联的蛋白质，其在过量CQ的存在下被抑制。使用MS-CETSA，我们确定了83个候选相互作用的蛋白质，共3375测量寄生虫蛋白质。同时，我们确定了8个蛋白质作为最潜在的命中，这是通常由两种方法确定。我们发现CQ可以通过直接与一些关键酶结合来破坏疟原虫的糖酵解和能量代谢，这是一种不同于其众所周知的疟原虫色素形成抑制作用的新机制。这是第一个报告，确定CQ抗疟靶标的标记（ABPP）和无标记（MS-CETSA）方法的平行使用。在线版本包含补充材料，可通过10.1186/s40779-022-00390-3获得。
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. 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. 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. 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. The online version contains supplementary material available at 10.1186/s40779-022-00390-3.
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影响因子：
3.5
作者：
Mehta, M;Sonawat, HM;Sharma, S
通讯作者：
Sharma, S
DOI：
10.1073/pnas.1910123116
发表时间：
2019-11-12
影响因子：
11.1
作者：
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通讯作者：
Als-Nielsen, Jens
影响因子：
81.5
作者：
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通讯作者：
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影响因子：
12
作者：
Chan, Maurice;Tan, Doreen S H;Sim, T S
通讯作者：
Sim, T S
影响因子：
11.5
作者：
Ashley EA;Phyo AP
通讯作者：
Phyo AP