The bio-layer interferometry (BLI) assay is a valuable tool for measuring protein-protein and protein-small molecule interactions. Here, we first describe the application of this novel label-free technique to study the interaction of human EAG1 (hEAG1) channel proteins with the small molecule PIP2. hEAG1 channel has been recognized as potential therapeutic target because of its aberrant overexpression in cancers and a few gain-of-function mutations involved in some types of neurological diseases. We purified hEAG1 channel proteins from a mammalian stable expression system and measured the interaction with PIP2 by BLI. The successful measurement of the kinetics of binding between hEAG1 protein and PIP2 demonstrates that the BLI assay is a potential high-throughput approach used for novel small-molecule ligand screening in ion channel pharmacology.
基金：
Bio-ID Center; SJTU Cross-Disciplinary Research Fund in Medicine and Engineering [YG2016QN66]; National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31271217]; National Basic Research Program of China[2014CB910304]
第一作者机构：[1]Shanghai Jiao Tong Univ, Tongren Hosp, Sch Med, Dept Gen Surg, Shanghai, Peoples R China
通讯作者：
推荐引用方式(GB/T 7714)：
Han Bo,Zhang Man,Sun Peng,et al.Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay[J].JOVE-JOURNAL OF VISUALIZED EXPERIMENTS.2018,(133):doi:10.3791/56846.
APA：
Han, Bo,Zhang, Man,Sun, Peng&Hou, Shangwei.(2018).Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay.JOVE-JOURNAL OF VISUALIZED EXPERIMENTS,,(133)
MLA：
Han, Bo,et al."Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay".JOVE-JOURNAL OF VISUALIZED EXPERIMENTS ..133(2018)
