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FIBS-enabled Noninvasive Metabolic Profiling
Published on: February 3, 2014
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Monitoring glycolytic dynamics in single cells using a fluorescent biosensor for fructose 1,6-bisphosphate
John N Koberstein1, Melissa L Stewart1, Chadwick B Smith1
1Vollum Institute, Oregon Health & Science University, Portland, OR 97239.
Summary
Scientists developed HYlight, a fluorescent biosensor for fructose 1,6-bisphosphate (FBP), to track cellular glycolysis in real-time. This tool reveals new insights into beta-cell metabolic dynamics and heterogeneity.
Area of Science:
- Cellular metabolism
- Biochemistry
- Molecular biology
Background:
- Cellular metabolism requires precise spatial and temporal regulation of energy production and consumption.
- Fluorescent biosensors offer real-time, single-cell resolution for monitoring metabolite levels.
- Fructose 1,6-bisphosphate (FBP) is a key indicator of glycolytic flux.
Purpose of the Study:
- To develop a novel fluorescent biosensor for real-time monitoring of FBP.
- To investigate cellular metabolism and glycolysis dynamics in living cells and tissues.
- To uncover novel aspects of beta-cell glycolytic heterogeneity.
Main Methods:
- Engineered a fluorescent biosensor (HYlight) by inserting GFP into the CggR protein from Bacillus subtilis.
- Utilized HYlight to monitor FBP levels in real-time.
- Applied various metabolic and pharmacological perturbations to living cells and tissues.
Main Results:
- HYlight reliably reports real-time glycolytic dynamics.
- The sensor functions effectively under diverse metabolic and pharmacological conditions.
- Uncovered previously unknown heterogeneity and dynamics in beta-cell glycolysis.
Conclusions:
- HYlight is a robust tool for studying glycolysis and cellular metabolism.
- The sensor enables new discoveries regarding metabolic regulation in various biological contexts.
- Revealed novel insights into beta-cell metabolic heterogeneity and dynamics.
