Interaction of Taspine Derivative TPD7 with Vascular Endothelial Growth Factor Receptor 2 by Cell Membrane Chromatography
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Affinity chromatography, as one of the important tools for studying the binding affinity between ligands and receptors, plays a key role in the process of screening and analyzing active compounds. Cell membrane chromatography (CMC) utilizes cell membrane receptors as stationary phases to perform screening and binding affinity of compounds with specific receptors whose three-dimensional configurations and biological activities were largely retained. In this study, a highly vascular endothelial growth factor receptor 2 (VEGFR2) expressing CMC method was established to investigate the binding affinity between TPD7 and VEGFR2. Competitive binding study taking sunitinib malate as the marker was used to inspect the binding site of TPD7 on VEGFR2. Results showed that TPD7 shared the same binding site with sunitinib on VEGFR2, which was consistent with the results of molecular docking. The equilibrium dissociation constants (KD) of TPD7 was (0.29 ± 0.02) × 10−6 M. Furthermore, TPD7 could alter the VEGFR2 kinase and significantly decrease phosphorylation of VEGFR2 in a dose-dependent manner. The studies showed that TPD7 could bind to VEGFR2 and then down-regulate the phosphorylation of VEGFR2.
KeywordsCell membrane chromatography Competitive binding Equilibrium dissociation constant VEGFR2 TPD7
This study was supported by National Natural Science Foundation of China (Grant no. 81603148), the National Science Foundation for Post-doctoral Scientists of China (Grant no. 2016M592806), and the Fundamental Research Funds for the Central Universities (Grant no. xzy012019078).
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Conflict of interest
The authors declare that there are no conflicts of interest.
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