Investigation of the binding characteristics between ligands and epidermal growth factor receptor by cell membrane chromatography
Investigation of the binding characteristics between ligands and epidermal growth factor receptor by cell membrane chromatography
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细胞膜色谱法研究配体与表皮生长因子受体的结合特性
DOI：
10.1002/jmr.2701
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发表时间：
2018-06-01
影响因子：
2.7
通讯作者：
Ma, Weina
中科院分区：
文献类型：
--
作者：
Yang, Liu;Zhu, Man;Ma, Weina
配体与其受体之间的结合性质对于许多生物过程是非常重要的。在本研究中，我们开发了一种高表皮生长因子受体（EGFR）表达细胞膜色谱（CMC）方法，以研究EGFR与配体吉非替尼、厄洛替尼、canertinib、阿法替尼和凡德他尼之间的结合特征。使用吉非替尼作为标记物的竞争结合分析用于研究在EGFR上的特异性结合位点发生的相互作用。从HEK 293-EGFR/CMC柱测量吉非替尼对这些配体占据的结合位点上的置换能力，其显示以下顺序：gefitinib（K-D，K- 8.49 +/- 0.11x10（-7）M）>厄洛替尼（K-D，K- 1.07 +/- 0.02x10（-6）M）>卡奈替尼（K-D，K- 1.41 +/- 0.07x10（-6）M）>阿法替尼（K-D，K- 1.80 +/- 0.12x10（-6）M）>凡德他尼（K-D，K- 1.99 +/- 0.03x10（-6）M）。这一顺序对应于由前沿位移分析估计的值和分子对接获得的分数。热力学分析表明，EGFR与5种配体结合的主要作用力为氢键或货车范德华力。总之，这些结果表明，CMC方法可以是一种有效的工具，研究配体和受体之间的结合特性。
The binding property between a ligand and its receptor is very important for numerous biological processes. In this study, we developed a high epidermal growth factor receptor (EGFR)-expression cell membrane chromatography (CMC) method to investigate the binding characteristics between EGFR and the ligands gefitinib, erlotinib, canertinib, afatinib, and vandetanib. Competitive binding analysis using gefitinib as the marker was used to investigate the interactions that occurred at specific binding sites on EGFR. The ability of displacement was measured from the HEK293-EGFR/CMC column on the binding sites occupied by gefitinib for these ligands, which revealed the following order: gefitinib (K-D,K- 8.49 +/- 0.11x10(-7)M)>erlotinib (K-D,K- 1.07 +/- 0.02x10(-6)M)>canertinib (K-D,K- 1.41 +/- 0.07x10(-6)M)>afatinib (K-D,K- 1.80 +/- 0.12x10(-6)M)>vandetanib (K-D,K- 1.99 +/- 0.03x10(-6)M). This order corresponded with the values estimated by frontal displacement analysis and the scores obtained with molecular docking. Furthermore, thermodynamic analysis indicated that the hydrogen bond or Van der Waals force was the main interaction force in the process of EGFR binding to all 5 ligands. Overall, these results demonstrate that a CMC method could be an effective tool to investigate the binding characteristics between ligands and receptors.