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A Sensitive Visual Method for the Detection of Hydrogen Sulfide Producing Bacteria
Published on: June 27, 2022
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A highly responsive and selective fluorescent probe for imaging physiological hydrogen sulfide.
1Department of Chemistry, University of California , 501 Big Springs Road, Riverside, California 92521, United States.
Biochemistry
|August 21, 2014
Summary
Researchers developed hsGFP, a novel genetically encoded probe, to detect hydrogen sulfide (H2S) in biological systems. This highly sensitive and selective probe enables real-time imaging of H2S in living cells and subcellular compartments.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Imaging
Background:
- Hydrogen sulfide (H2S) is recognized as a crucial gasotransmitter involved in cellular signaling and various pathophysiological processes.
- Accurate detection of H2S in biological environments is essential for understanding its roles.
- Existing analytical methods for H2S detection face limitations in sensitivity, selectivity, or applicability to living systems.
Purpose of the Study:
- To develop a novel, highly responsive, and selective genetically encoded probe for detecting hydrogen sulfide (H2S).
- To enable the imaging of H2S in vitro and within living mammalian cells with subcellular resolution.
- To validate the probe's performance in visualizing biologically relevant H2S production.
Main Methods:
- Development of a genetically encoded fluorescent probe, hsGFP, engineered for H2S detection.
- Characterization of hsGFP's fluorescence response, folding efficiency, and chromophore formation.
- Application of hsGFP for in vitro and in vivo imaging of H2S in mammalian cells, including subcellular localization (mitochondria, nuclei, ion channels).
- Utilizing hsGFP to visualize H2S generated enzymatically from L-cysteine in HEK293T cells.
Main Results:
- hsGFP demonstrated high responsiveness and selectivity for H2S detection.
- The probe exhibited efficient folding and chromophore formation, crucial for robust fluorescence signaling.
- hsGFP allowed for precise subcellular localization and imaging of H2S in living cells.
- The probe successfully visualized H2S production from L-cysteine in HEK293T cells.
Conclusions:
- hsGFP represents a significant advancement in genetically encoded probes for H2S detection.
- The probe's properties facilitate sensitive and selective H2S imaging in complex biological systems.
- hsGFP offers a valuable tool for investigating the physiological and pathological roles of H2S at the cellular level.
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