| ORIGINAL ARTICLES |
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Identification and verification of methylenetetrahydrofolate dehydrogenase 1-like protein as the binding target of natural product pseudolaric acid A |
| Haoqi Dong1,2, Xinni Yang1, Peiying Wang2,3, Weiya Huang2,3, Liang Zhang1, Song Song4, Jiangxin Liu1 |
1. State Key Laboratory of Phytochemistry and Natural Medicines, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650201, China;
2. University of Chinese Academy of Sciences, Beijing, 100049, China;
3. Yunnan University, Kunming, 650500, China;
4. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China |
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Abstract Natural product pseudolaric acid A (PAA), the main bioactive component from Traditional Chinese Medicine Pseudolarix cortex (“tujingpi”), is a promising anticancer agent. However, its potential molecular targets are not clear and this hinders its development. In this study, chemical proteomics approaches including activity-based protein profiling (ABPP) and drug affinity responsive target stability (DARTS) technology, followed by quantitative proteomics, were combined to reveal the target of PAA. Target validation was performed by NMR techniques and surface plasmon resonance. Methylenetetrahydrofolate dehydrogenase 1-like (MTHFD1L) was identified and further confirmed to be the target of PAA. The direct interaction and binding mode between MTHFD1L and PAA were elaborated. PAA induced the accumulation of the reactive oxygen species (ROS) which mediates the antitumor effect. Transcriptome and network pharmacology analysis reveals the effects of PAA on the gene expressions of the associated pathways. Taken together, our findings proposed a new target that could be used for structure-based rational design and modifications of PAA.
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| Keywords
Natural product
Target identification
Chemical proteomics
Target verification
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| Fund:This work was supported by Yunnan Fundamental Research Projects (Grant No. 202201AT070183), Biological Medicine Special Project of Yunnan (202402AA310015), the Ten thousand-talents program of Yunnan Province to Jiangxin Liu, and the State Key Laboratory of Natural and Biomimetic Drugs, Peking University. |
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Issue Date: 18 June 2025
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