用于乳酸成像的亮红色荧光遗传编码传感器。
A bright red fluorescent genetically encoded sensor for lactate imaging.
机构信息
Optogenetics & Synthetic Biology Interdisciplinary Research Center, Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism, State Key Laboratory of Bioreactor Engineering, School of Pharmacy, East China University of Science and Technology, Shanghai, 200237, China.
Optogenetics & Synthetic Biology Interdisciplinary Research Center, Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism, State Key Laboratory of Bioreactor Engineering, School of Pharmacy, East China University of Science and Technology, Shanghai, 200237, China; Research Unit of New Techniques for Live-cell Metabolic Imaging, Chinese Academy of Medical Sciences, Beijing, 100730, China.
出版信息
Biochem Biophys Res Commun. 2024 Nov 19;734:150449. doi: 10.1016/j.bbrc.2024.150449. Epub 2024 Jul 25.
Lactate plays a crucial role in energy metabolism and greatly impacts protein activities, exerting diverse physiological and pathological effects. Therefore, convenient lactate assays for tracking spatiotemporal dynamics in living cells are desirable. In this paper, we engineered and optimized a red fluorescent protein sensor for l-lactate named FiLa-Red. This indicator exhibited a maximal fluorescence change of 730 % and an apparent dissociation constant (K) of approximately 460 μM. By utilizing FiLa-Red and other sensors, we monitored energy metabolism in a multiplex manner by simultaneously tracking lactate and NAD/NADH abundance in the cytoplasm, nucleus, and mitochondria. The FiLa-Red sensor is expected to be a useful tool for performing metabolic analysis in vitro, in living cells and in vivo.
乳酸在能量代谢中起着至关重要的作用，极大地影响蛋白质的活性，发挥着多种生理和病理作用。因此，理想情况下，人们希望有一种方便的乳酸检测方法，用于跟踪活细胞中的时空动态。在本文中，我们设计并优化了一种名为 FiLa-Red 的用于 l-乳酸的红色荧光蛋白传感器。该指示剂的最大荧光变化为 730%，表观解离常数（K）约为 460 μM。通过利用 FiLa-Red 和其他传感器，我们可以通过同时跟踪细胞质、细胞核和线粒体中乳酸和 NAD/NADH 的丰度，以多重方式监测能量代谢。FiLa-Red 传感器有望成为体外、活细胞和体内代谢分析的有用工具。
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