Interaction of Taspine Derivative TPD7 with Vascular Endothelial Growth Factor Receptor 2 by Cell Membrane Chromatography
Interaction of Taspine Derivative TPD7 with Vascular Endothelial Growth Factor Receptor 2 by Cell Membrane Chromatography
复制标题
通过细胞膜色谱法研究 Taspine 衍生物 TPD7 与血管内皮生长因子受体 2 的相互作用
DOI：
10.1007/s10337-019-03801-1
复制
发表时间：
2019-12-01
期刊：
影响因子：
1.7
通讯作者：
Ma，Weina
中科院分区：
文献类型：
--
作者：
Yang，Liu;Zeng，Yingnan;Ma，Weina
摘要亲和层析是研究配体与受体结合亲和力的重要工具之一，在活性化合物的筛选和分析过程中起着关键作用。细胞膜色谱法（CMC）利用细胞膜受体作为固定相，筛选化合物与特定受体的结合亲和力，这些受体的三维构型和生物活性在很大程度上被保留。本研究建立了一种高表达血管内皮生长因子受体2 （VEGFR2）的CMC方法，研究TPD7与VEGFR2的结合亲和力。采用以苹果酸舒尼替尼为标记物的竞争结合研究，检测TPD7在VEGFR2上的结合位点。结果显示TPD7与舒尼替尼在VEGFR2上有相同的结合位点，这与分子对接的结果一致。TPD7的平衡解离常数（KD）为（0.29±0.02）× 10−6M。此外，TPD7可以改变VEGFR2激酶，并以剂量依赖性的方式显著降低VEGFR2的磷酸化。研究表明，TPD7可以结合VEGFR2，进而下调VEGFR2的磷酸化水平。图形抽象
AbstractAffinity 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) × 10−6M. 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.Graphic Abstract