Experimental Molecular Biophysics, Department of Physics, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, Germany.
Institute of Molecular Science, Universitat de Valencia, Catedrático José Beltrán Martínez, No. 2, 46980 Paterna, Spain.
J Chem Phys. 2022 May 28;156(20):204201. doi: 10.1063/5.0088526.
Mid-IR spectroscopy is a powerful and label-free technique to investigate protein reactions. In this study, we use quantum-cascade-laser-based dual-comb spectroscopy to probe protein conformational changes and protonation events by a single-shot experiment. By using a well-characterized membrane protein, bacteriorhodopsin, we provide a comparison between dual-comb spectroscopy and our homebuilt tunable quantum cascade laser (QCL)-based scanning spectrometer as tools to monitor irreversible reactions with high time resolution. In conclusion, QCL-based infrared spectroscopy is demonstrated to be feasible for tracing functionally relevant protein structural changes and proton translocations by single-shot experiments. Thus, we envisage a bright future for applications of this technology for monitoring the kinetics of irreversible reactions as in (bio-)chemical transformations.
中红外光谱是一种强大的、无需标记的技术，可用于研究蛋白质反应。在这项研究中，我们使用基于量子级联激光器的双梳光谱学，通过单次实验来探测蛋白质构象变化和质子化事件。通过使用一种经过良好表征的膜蛋白——菌紫质，我们将双梳光谱学与我们自制的基于可调谐量子级联激光器的扫描光谱仪进行了比较，作为监测具有高时间分辨率的不可逆反应的工具。总之，基于 QCL 的红外光谱学被证明可通过单次实验来追踪与功能相关的蛋白质结构变化和质子迁移。因此，我们期望这项技术在监测（生物）化学转化等不可逆反应动力学方面具有广阔的应用前景。
