Manchester Institute of Biotechnology and Department of Chemistry, School of Natural Sciences, Faculty of Science and Engineering, The University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K.
Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Harwell Oxford, Didcot OX11 0QX, United Kingdom.
Biochemistry. 2020 Aug 18;59(32):2909-2915. doi: 10.1021/acs.biochem.0c00382. Epub 2020 Aug 6.
Due to the recent advances in X-ray free electron laser techniques, bilin-containing cyanobacteriochrome photoreceptors have become prime targets for the ever-expanding field of time-resolved structural biology. However, to facilitate these challenging studies, it is essential that the time scales of any structural changes during the photocycles of cyanobacteriochromes be established. Here, we have used visible and infrared transient absorption spectroscopy to probe the photocycle of a model cyanobacteriochrome system, TePixJ. The kinetics span multiple orders of magnitude from picoseconds to seconds. Localized changes in the bilin binding pocket occur in picoseconds to nanoseconds, followed by more large-scale changes in protein structure, including formation and breakage of a second thioether linkage, in microseconds to milliseconds. The characterization of the entire photocycle will provide a vital frame of reference for future time-resolved structural studies of this model photoreceptor.
由于 X 射线自由电子激光技术的最新进展,含双吡咯环的蓝藻光感受器已成为时间分辨结构生物学这一不断扩展领域的主要研究目标。然而,为了促进这些具有挑战性的研究,确定在蓝藻光感受器的光循环过程中任何结构变化的时间尺度是至关重要的。在这里,我们使用可见和红外瞬态吸收光谱来探测模型蓝藻光感受器系统 TePixJ 的光循环。动力学范围跨越从皮秒到秒的多个数量级。在皮秒到纳秒的时间内,双吡咯环结合口袋中发生局部变化,随后在微秒到毫秒的时间内,蛋白质结构发生更大规模的变化,包括形成和破坏第二个硫醚键。整个光循环的特性将为未来对该模型光感受器进行时间分辨结构研究提供重要的参考框架。