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Glutamine Flux Imaging Using Genetically Encoded Sensors

Glutamine Flux Imaging Using Genetically Encoded Sensors

Published on: July 31, 2014

A bright red fluorescent genetically encoded sensor for lactate imaging.

Xuanming Chang¹, Xiaoqian Chen¹, Xiuze Zhang¹

¹Optogenetics & 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.

Biochemical and Biophysical Research Communications

|August 3, 2024

Summary

Scientists developed FiLa-Red, a novel red fluorescent sensor for tracking lactate in living cells. This tool enables precise monitoring of cellular energy metabolism and protein activity, advancing diagnostic capabilities.

Keywords:

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

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Area of Science:

Biochemistry
Cell Biology
Molecular Imaging

Background:

Lactate is vital in cellular energy metabolism, influencing protein activity and various physiological and pathological processes.
Accurate spatiotemporal tracking of lactate dynamics in living systems is essential for understanding cellular functions.
Existing lactate assays lack the convenience and precision required for real-time cellular analysis.

Purpose of the Study:

To engineer and optimize a red fluorescent protein sensor for sensitive and specific detection of l-lactate.
To characterize the performance of the novel lactate sensor, FiLa-Red.
To demonstrate the utility of FiLa-Red in multiplexed monitoring of cellular energy metabolism.

Main Methods:

Development and optimization of a red fluorescent protein-based sensor for l-lactate.
Characterization of sensor performance, including fluorescence change and dissociation constant (K_d).
Multiplexed imaging of lactate, NAD+/NADH in cellular compartments (cytoplasm, nucleus, mitochondria) using FiLa-Red and other sensors.

Main Results:

Engineered FiLa-Red sensor exhibits a maximal fluorescence change of 730% for l-lactate.
The sensor has an apparent dissociation constant (K_d) of approximately 460 μM, indicating high affinity.
Successful multiplexed monitoring of lactate and NAD+/NADH in various subcellular locations was achieved.

Conclusions:

FiLa-Red is a highly sensitive and effective red fluorescent sensor for l-lactate detection.
The sensor facilitates real-time analysis of cellular energy metabolism in vitro, in living cells, and potentially in vivo.
FiLa-Red represents a valuable advancement for metabolic research and diagnostics.