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Updated: Jul 9, 2025

Detection of Nitric Oxide and Superoxide Radical Anion by Electron Paramagnetic Resonance Spectroscopy from Cells using Spin Traps
Published on: August 18, 2012
Characterization of an N-Allylglyoxylamide-Based Bioorthogonal Nitrone Trap
Daniel Lee1, Simon Latour2,3, Michael Emblem1
1Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario M5S 3H6, Canada.
This study introduces a new bioorthogonal reaction combining nitrones and aldehydes to form stable bicyclic isoxazolidines. This efficient reaction works in cell lysates and on cell surfaces, expanding bioorthogonal chemistry tools.
Area of Science:
- Chemical Biology
- Organic Chemistry
- Bioconjugation
Background:
- Aldehydes and nitrones are valuable in bioorthogonal chemistry for creating irreversible conjugates.
- Existing bioorthogonal reactions include those involving strained alkynes and nitrones.
Purpose of the Study:
- To develop a novel bioorthogonal reaction utilizing the reactivity of nitrones and aldehydes.
- To create a cascade reaction forming stable bicyclic isoxazolidines.
- To assess the reaction's utility in biological systems and its compatibility with other click chemistry.
Main Methods:
- A cascade reaction between an N-allylglyoxylamide and an N-alkylhydroxylamine was employed.
- The reaction was catalyzed by 5-methoxyanthranilic acid under physiological conditions (pH 7).
- The reaction's bioorthogonality was tested in cell lysates and on the surface of HEK293 cells using HaloTag7 protein.
Main Results:
- A novel bicyclic isoxazolidine was successfully synthesized via a cascade reaction.
- The reaction demonstrated favorable kinetics and was efficiently catalyzed by 5-methoxyanthranilic acid at pH 7.
- The reaction proved bioorthogonal in complex cell lysates and functional on cell surfaces.
- Compatibility with strain-promoted alkyne-azide click reactions was confirmed.
Conclusions:
- A new, efficient, and bioorthogonal reaction between aldehydes and nitrones has been established.
- This reaction forms stable bicyclic isoxazolidines and is suitable for applications in chemical biology.
- The developed chemistry expands the toolkit for bioconjugation and bioimaging.
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