Lehrstuhl für Biophysik, Ruhr-Universität Bochum, 44780, Bochum, Germany.
Molecular Interactions and Biophysics, Merck KGaA, Frankfurter Strasse 250, 64293, Darmstadt, Germany.
Angew Chem Int Ed Engl. 2018 Jul 26;57(31):9955-9960. doi: 10.1002/anie.201802603. Epub 2018 Jul 3.
Investigation of protein-ligand interactions is crucial during early drug-discovery processes. ATR-FTIR spectroscopy can detect label-free protein-ligand interactions with high spatiotemporal resolution. Here we immobilized, as an example, the heat shock protein HSP90 on an ATR crystal. This protein is an important molecular target for drugs against several diseases including cancer. With our novel approach we investigated a ligand-induced secondary structural change. Two specific binding modes of 19 drug-like compounds were analyzed. Different binding modes can lead to different efficacy and specificity of different drugs. In addition, the k values of ligand dissociation were obtained. The results were validated by X-ray crystallography for the structural change and by SPR experiments for the dissociation kinetics, but our method yields all data in a single and simple experiment.
在药物发现早期过程中，研究蛋白质-配体相互作用至关重要。ATR-FTIR 光谱法可在高时空分辨率下检测无标记的蛋白质-配体相互作用。在这里，我们以热休克蛋白 HSP90 为例将其固定在 ATR 晶体上。该蛋白是针对包括癌症在内的多种疾病的药物的重要分子靶标。通过我们的新方法，我们研究了配体诱导的二级结构变化。分析了 19 种类似药物的两种特定结合模式。不同的结合模式可能导致不同药物的不同疗效和特异性。此外，还获得了配体解离的 k 值。结果通过 X 射线晶体学验证了结构变化，通过 SPR 实验验证了解离动力学，但我们的方法在单次简单的实验中即可获得所有数据。
