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H2O2 and Phosphorylated Peptide Dual-Responsive Nanochannel Device
Lang Peng1,2, Cunli Wang2, Qiwen Lin3
1College of Chemistry and Chemical Engineering, Wuhan Textile University, 1 Sunshine Road, Wuhan, Hubei 430200, P. R. China.
Analytical Chemistry
|June 26, 2023
Summary
This study presents a novel nanochannel device that simultaneously detects hydrogen peroxide (H₂O₂) and phosphorylated peptides (PPs). This dual-sensing capability offers a new tool for studying cellular signaling and screening kinase inhibitors.
Area of Science:
- Biotechnology
- Biosensing
- Cellular Signaling
Background:
- Oxidation and protein phosphorylation are key regulators of cellular processes.
- Oxidative stress can alter kinase and phosphatase activity, impacting protein phosphorylation.
- Simultaneous detection of oxidation and phosphorylation is challenging but crucial for understanding cellular regulation.
Purpose of the Study:
- To develop a proof-of-concept nanochannel device for simultaneous detection of H₂O₂ and phosphorylated peptides (PPs).
- To create a peptide-based sensor for monitoring cellular responses to stimuli like platelet-derived growth factor (PDGF).
Main Methods:
- Immobilization of a specifically designed peptide (GGGCEG(GPGGA)₄CEGRRRR) onto the inner walls of conical nanochannels.
- Utilizing the H₂O₂-induced random coil-to-α-helix transition and PP binding-induced charge shielding for dual sensing.
- Measuring changes in transmembrane ionic current in response to H₂O₂ and PPs.
Main Results:
- The nanochannel device demonstrated sensitive and dual-responsive detection of both H₂O₂ and PPs.
- The device successfully detected reactive oxygen species and changes in PP levels in PDGF-stimulated 3T3-L1 cells.
- Real-time monitoring of kinase activity was achieved, showing potential for inhibitor screening.
Conclusions:
- The developed peptide-modified nanochannel device offers a novel platform for simultaneous sensing of oxidation and protein phosphorylation.
- This technology has significant potential for applications in cellular signaling research and drug discovery, particularly for kinase inhibitor screening.
