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  3. A bright red fluorescent genetically encoded sensor for lactate imaging

A bright red fluorescent genetically encoded sensor for lactate imaging

  • Biochem Biophys Res Commun. 2024 Jul 25:734:150449. doi: 10.1016/j.bbrc.2024.150449.
Xuanming Chang 1,  Xiaoqian Chen 1,  Xiuze Zhang 1,  Nian Chen 1,  Weitao Tang 1,  Zhuo Zhang 1,  Sulin Zheng 1,  Jiong Huang 1,  Yihan Ji 1,  Yuzheng Zhao 2,  Yi Yang 2,  Xie Li 3
Affiliations

Affiliations

  • 1 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.
  • 2 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.
  • 3 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. Electronic address: xieli@ecust.edu.cn.
PMID: 39096623 DOI: 10.1016/j.bbrc.2024.150449
Abstract

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 (Kd) 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.

Keywords

Fluorescence imaging; Genetically encoded fluorescent sensor; Lactate metabolism; Real-time monitoring; Spatiotemporal dynamics.

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