Department of Chemistry, Aarhus University , DK-8000 Aarhus, Denmark.
Department of Molecular Biology and Genetics, Aarhus University , DK-8000 Aarhus, Denmark.
J Phys Chem B. 2017 Oct 12;121(40):9366-9371. doi: 10.1021/acs.jpcb.7b07831. Epub 2017 Sep 27.
Optogenetics has been, and will continue to be, a boon to mechanistic studies of cellular processes. Genetically encodable proteins that sensitize the production of reactive oxygen species (ROS) are expected to play an increasingly important role, particularly in elucidating mechanisms of temporally and spatially dependent cell signaling. However, a substantial challenge in developing such photosensitizing proteins has been to funnel the optical excitation energy into the initial selective production of only one ROS. Singlet molecular oxygen, O(aΔ), is a ROS known to have a wide range of effects on cell function. Nevertheless, mechanistic details of singlet oxygen's behavior in a cell are lacking. On the basis of the rational optimization of a LOV-derived flavoprotein, we now report the development and photophysical characterization of a protein-encased photosensitizer that efficiently and selectively produces singlet oxygen at the expense of other ROS, especially ROS that derive from photoinduced electron transfer reactions. These results set the stage for a plethora of new experiments to elucidate ROS-mediated events in cells.
光遗传学一直是、并且将继续是细胞过程机械研究的福音。预计能够敏感地产生活性氧（ROS）的基因可编码蛋白将发挥越来越重要的作用，尤其是在阐明随时间和空间变化的细胞信号转导机制方面。然而，开发这种光敏蛋白的一个重大挑战是将光学激发能量引导到仅选择性地产生一种 ROS。单线态氧（O(aΔ)）是一种 ROS，已知其对细胞功能有广泛的影响。然而，在细胞中单线态氧行为的机制细节尚不清楚。基于对 LOV 衍生黄素蛋白的合理优化，我们现在报告了一种蛋白质包裹的光敏剂的开发和光物理特性表征，该光敏剂能够有效地、选择性地产生单线态氧，而不会产生其他 ROS，特别是不会产生来自光诱导电子转移反应的 ROS。这些结果为大量新的实验奠定了基础，这些实验旨在阐明细胞中 ROS 介导的事件。
