Imdea Nanociencia, Madrid, Spain.
Physics of Biological Function, Institut Pasteur, Paris, France.
Methods Mol Biol. 2022;2376:117-134. doi: 10.1007/978-1-0716-1716-8_6.
The development of ultrafast kinetic methods is one of the factors that allowed the research on protein folding to flourish over the last 20 years. The introduction of new optical triggering techniques enabled to experimentally investigate the protein dynamics at the nanosecond to millisecond timescale, allowing researchers to test theoretical predictions and providing experimental benchmarks for computer simulations. In this work, the details of how to perform kinetic experiments by the laser-induced temperature-jump technique, using the two most commonly used probing techniques (namely infrared absorption and fluorescence spectroscopy) are given, with a strong emphasis on the practical details.
超快动力学方法的发展是促使过去 20 年来蛋白质折叠研究蓬勃发展的因素之一。新型光学触发技术的引入使得能够在纳秒到毫秒时间尺度上对蛋白质动力学进行实验研究,从而使研究人员能够检验理论预测,并为计算机模拟提供实验基准。在这项工作中,给出了如何使用两种最常用的探测技术(即红外吸收和荧光光谱学)通过激光诱导温度跃变技术进行动力学实验的细节,重点介绍了实际细节。