Activation of visual rhodopsin probed by single-shot transient IR spectroscopy

In order to understand the structure-function relationships of proteins, it is important to study their dynamics under physiological conditions. The advent of X-ray free electron lasers has made it possible to obtain the three-dimensional structures of proteins and their reaction intermediates at room temperature. However, these experiments are very demanding and require extensive planning. Here, we demonstrate that time-resolved infrared difference spectroscopy using quantum cascade lasers is a powerful tool for studying the dynamics of protein conformational changes. This method complements structural biology experiments and aids in data interpretation. We demonstrate the feasibility of studying the irreversible photoreaction of the G-protein-coupled receptor rhodopsin using single shots.