Chemphyschem. 2014 Mar 17;15(4):671-6. doi: 10.1002/cphc.201300755.
Protein–ligand interactions play an important role in many biological processes. Notably, membrane receptors are the starting point for a huge variety of cellular signal transduction pathways. Quantifying the binding affinity of a ligand for its transmembrane receptor is of great importance as it provides information on the potency of the ligand. We developed a new experimental procedure to determine binding affinities of ligands for their membrane receptors directly on intact single cells using super-resolution imaging. Dissociation constants were determined by titrating fluorophore-labelled ligand against cells expressing the target protein and applying single-molecule imaging.
蛋白质-配体相互作用在许多生物过程中起着重要作用。值得注意的是，膜受体是各种细胞信号转导途径的起点。定量配体与其跨膜受体的结合亲和力非常重要，因为它提供了配体效力的信息。我们开发了一种新的实验程序，使用超分辨率成像直接在完整的单个细胞上测定配体与其膜受体的结合亲和力。通过将荧光标记的配体滴定到表达靶蛋白的细胞中，并应用单分子成像来确定离解常数。
