Q-SHINE: A versatile sensor for glutamine measurement via ligand-induced dimerization

Lim, Yun; Kim, Ji Yul; Jung, Youn Hee; Lee, Jae Hoon; Baek, Min Seok; Jung, Jee-Hyun; Kim, Ho-Youn; Lee, Wookbin; Park, Keunwan; Seo, Myung Won

doi:10.1016/j.snb.2023.133951

“…Sensors 2024, 24, 3073 2 of 13 Conventional medical lab tests for glutamine, such as high-performance liquid chromatography (HPLC), have limited portability, high costs, and slow result times, which restrict patient access to treatment-monitoring tests [29,30]. This has motivated efforts to develop alternative glutamine biosensors utilizing a variety of biochemical and analytical methods [31][32][33][34][35][36][37]. Lim et al developed the Q-SHINE assay, in which a split glutaminebinding protein recombines with a split reporter protein that dimerizes upon binding of glutamine to produce a signal [31].…”

Section: Introductionmentioning

confidence: 99%

“…This has motivated efforts to develop alternative glutamine biosensors utilizing a variety of biochemical and analytical methods [31][32][33][34][35][36][37]. Lim et al developed the Q-SHINE assay, in which a split glutaminebinding protein recombines with a split reporter protein that dimerizes upon binding of glutamine to produce a signal [31]. This exciting study presents a unique and effective way to detect glutamine, though this is a relatively slow assay that requires several hours to obtain results, and the assay has not been demonstrated in a portable form.…”

Section: Introductionmentioning

confidence: 99%

“…Tanigawa et al harnessed cellular nutrient-response mechanisms to create an in vitro glutamine sensor, discovering novel protein-glutaminebinding interactions [36]. While these and other novel methods to detect glutamine [31][32][33][34][35][36][37] represent important progress in the field of glutamine sensing, no commercialized, portable glutamine tests are available. Thus, patients suffering from cancer and other diseases do not have laboratory equipment nor expertise to conduct frequent testing.…”

Section: Introductionmentioning

confidence: 99%

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Engineering the Signal Resolution of a Paper-Based Cell-Free Glutamine Biosensor with Genetic Engineering, Metabolic Engineering, and Process Optimization

Free,

Talley,

Hyer

et al. 2024

Sensors

0

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Specialized cancer treatments have the potential to exploit glutamine dependence to increase patient survival rates. Glutamine diagnostics capable of tracking a patient’s response to treatment would enable a personalized treatment dosage to optimize the tradeoff between treatment success and dangerous side effects. Current clinical glutamine testing requires sophisticated and expensive lab-based tests, which are not broadly available on a frequent, individualized basis. To address the need for a low-cost, portable glutamine diagnostic, this work engineers a cell-free glutamine biosensor to overcome assay background and signal-to-noise limitations evident in previously reported studies. The findings from this work culminate in the development of a shelf-stable, paper-based, colorimetric glutamine test with a high signal strength and a high signal-to-background ratio for dramatically improved signal resolution. While the engineered glutamine test is important progress towards improving the management of cancer and other health conditions, this work also expands the assay development field of the promising cell-free biosensing platform, which can facilitate the low-cost detection of a broad variety of target molecules with high clinical value.

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GlutaR: A High‐Performance Fluorescent Protein‐Based Sensor for Spatiotemporal Monitoring of Glutamine Dynamics In Vivo

Liu,

Zhao,

Wang

et al. 2024

Angewandte Chemie

0

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Glutamine is the most abundant amino acid in human blood and muscle, and is integral to a wide variety of functions in cancer cells. However, the inability to monitor the subcellular distribution of glutamine in real‐time has obscured understanding of glutamine metabolism under physiological and pathological conditions. Here, we report the development of a genetically encoded fluorescent sensor and demonstrate how this GlnBP‐cpYFP fusion “GlutaR sensor” undergoes glutamine‐induced conformational changes reflected in detectable fluorescence responses. Obtained after iterative screening of approximately 1,600 variants, GlutaR exhibits a ratiometric readout, fast response kinetics, and high responsivity, and we demonstrate its selectivity for monitoring glutamine fluctuations in multiple cell types. Additionally, using digitonin permeabilization of GlutaR HeLa cells, we generated a calibration curve and performed in situ titration to quantify free glutamine concentrations in subcellular compartments (cytosol, nucleus, mitochondria). Subsequently, we applied GlutaR to investigate how chemical and genetic inhibition of GS and GLS differentially alter glutamine levels in subcellular compartments. Finally, we demonstrate GlutaR’s ability to monitor dynamic glutamine levels in muscle and liver tissues of diabetic mice in vivo. These findings collectively demonstrate GlutaR as a versatile tool for the spatiotemporal characterization of glutamine metabolism in living cells and tissues.

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GlutaR: A High‐Performance Fluorescent Protein‐Based Sensor for Spatiotemporal Monitoring of Glutamine Dynamics In Vivo

Liu,

Zhao,

Wang

et al. 2024

Angew Chem Int Ed

0

View full text Add to dashboard Cite

Glutamine is the most abundant amino acid in human blood and muscle, and is integral to a wide variety of functions in cancer cells. However, the inability to monitor the subcellular distribution of glutamine in real‐time has obscured understanding of glutamine metabolism under physiological and pathological conditions. Here, we report the development of a genetically encoded fluorescent sensor and demonstrate how this GlnBP‐cpYFP fusion “GlutaR sensor” undergoes glutamine‐induced conformational changes reflected in detectable fluorescence responses. Obtained after iterative screening of approximately 1,600 variants, GlutaR exhibits a ratiometric readout, fast response kinetics, and high responsivity, and we demonstrate its selectivity for monitoring glutamine fluctuations in multiple cell types. Additionally, using digitonin permeabilization of GlutaR HeLa cells, we generated a calibration curve and performed in situ titration to quantify free glutamine concentrations in subcellular compartments (cytosol, nucleus, mitochondria). Subsequently, we applied GlutaR to investigate how chemical and genetic inhibition of GS and GLS differentially alter glutamine levels in subcellular compartments. Finally, we demonstrate GlutaR’s ability to monitor dynamic glutamine levels in muscle and liver tissues of diabetic mice in vivo. These findings collectively demonstrate GlutaR as a versatile tool for the spatiotemporal characterization of glutamine metabolism in living cells and tissues.

show abstract

Q-SHINE: A versatile sensor for glutamine measurement via ligand-induced dimerization

Cited by 3 publications

References 43 publications

Engineering the Signal Resolution of a Paper-Based Cell-Free Glutamine Biosensor with Genetic Engineering, Metabolic Engineering, and Process Optimization

Engineering the Signal Resolution of a Paper-Based Cell-Free Glutamine Biosensor with Genetic Engineering, Metabolic Engineering, and Process Optimization

GlutaR: A High‐Performance Fluorescent Protein‐Based Sensor for Spatiotemporal Monitoring of Glutamine Dynamics In Vivo

GlutaR: A High‐Performance Fluorescent Protein‐Based Sensor for Spatiotemporal Monitoring of Glutamine Dynamics In Vivo

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