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Novel peroxiredoxin-based sensor for sensitive detection of hydrogen peroxide
Yuhong Yang1, Yao Zhou1, Jingyi Li1
1State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.
Biochemical and Biophysical Research Communications
|July 28, 2019
Summary
Researchers developed TScGP, a novel fluorescent biosensor for detecting hydrogen peroxide (H₂O₂) in living cells. This highly sensitive sensor offers rapid detection, advancing cellular imaging and diagnostics.
Area of Science:
- Cellular Biology
- Biochemistry
- Biotechnology
Background:
- Genetically encoded sensors are crucial for detecting hydrogen peroxide (H₂O₂), a key signaling molecule.
- Existing H₂O₂ biosensors require improvements in responsiveness and sensitivity for advanced cellular studies.
Purpose of the Study:
- To develop a novel, highly responsive, and sensitive fluorescent biosensor for detecting H₂O₂ in living cells.
- To utilize *E. coli* as a platform for creating this new H₂O₂ sensor.
Main Methods:
- Engineered a fluorescent sensor, TScGP, using circularly permuted YFP (cpYFP).
- Incorporated a redox relay system involving fungal peroxiredoxin A (PrxA) and mutated thioredoxin A (TrxA).
- Constructed the sensor with cpYFP sandwiched between PrxA and TrxA, forming a disulfide bond in response to H₂O₂.
Main Results:
- TScGP successfully detected exogenous H₂O₂ in the range of 0.5–5 μM with high selectivity.
- Achieved rapid detection of H₂O₂ within 30 seconds in *E. coli*.
- Demonstrated application by detecting intracellular H₂O₂ production induced by menadione.
Conclusions:
- The Prx-Trx combination serves as an effective sensing moiety for high-performance H₂O₂ biosensor design.
- TScGP represents a significant advancement in real-time cellular H₂O₂ monitoring.
- This novel sensor platform has potential applications in various biological and medical research fields.
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