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Imaging Approaches to Assessments of Toxicological Oxidative Stress Using Genetically-encoded Fluorogenic Sensors
Published on: February 7, 2018
Reaction-based genetically encoded fluorescent hydrogen sulfide sensors
Si Chen1, Zhi-jie Chen, Wei Ren
1Department of Chemistry, University of California, 501 Big Springs Road, Riverside, California 92521, USA.
Journal of the American Chemical Society
|May 31, 2012
Summary
Researchers developed genetically encoded fluorescent protein probes for detecting hydrogen sulfide (H2S), a toxic gas and signaling molecule. These probes offer sensitive and selective H2S detection in living cells using advanced imaging techniques.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- Hydrogen sulfide (H2S) is a crucial signaling molecule with diverse physiological roles.
- The selective and sensitive detection of H2S remains a significant analytical challenge.
- Existing methods for H2S detection often lack specificity or are unsuitable for live-cell applications.
Purpose of the Study:
- To develop novel genetically encoded fluorescent protein (FP)-based probes for selective H2S detection.
- To engineer FP chromophores with sulfide-reactive functional groups for enhanced fluorescence.
- To demonstrate the utility of these probes for monitoring H2S in living cells.
Main Methods:
- Genetic code expansion in E. coli and mammalian cells to incorporate modified amino acids.
- Chemical modification of FP chromophores with azide functional groups.
- Utilizing H2S-mediated reduction of azide for fluorescence signal generation.
- Spectroscopic and microscopic imaging for fluorescence detection.
- Engineering circularly permuted FPs for improved sensor performance.
Main Results:
- Genetically encoded FP probes capable of selective H2S detection were successfully developed.
- Modified FP chromophores showed H2S-dependent fluorescence enhancement.
- A circularly permuted FP-based sensor exhibited faster response times.
- The probes demonstrated feasibility for monitoring H2S in living mammalian cells.
Conclusions:
- Genetically encoded FP probes represent a powerful new tool for H2S detection.
- These probes enable sensitive and selective monitoring of H2S in biological systems.
- The developed technology has implications for understanding H2S biology and related diseases.
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