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Improved Photoinduced Fluorogenic Alkene-Tetrazole Reaction for Protein Labeling
Xin Shang1, Rui Lai1, Xi Song1
1Department of Chemistry, ‡Department of Chemistry, Nebraska Center for Materials and Nanoscience, and Center for Integrated Biomolecular Communication, and §Department of Chemical & Biomolecular Engineering, University of Nebraska-Lincoln , Lincoln, Nebraska 68588, United States.
Bioconjugate Chemistry
|October 13, 2017
Summary
Researchers improved fluorophore formation using bioorthogonal chemistry. The styrene-tetrazole reaction enhanced quantum yield for live-cell imaging applications.
Area of Science:
- Bioorthogonal Chemistry
- Chemical Biology
- Organic Synthesis
Background:
- The 1,3-dipolar cycloaddition between alkenes and tetrazoles is a rare bioorthogonal reaction forming fluorophores.
- This reaction is attractive for live-cell imaging due to minimal washing and potential for spatiotemporal resolution.
Purpose of the Study:
- To enhance the fluorogenic properties of the alkene-tetrazole reaction for improved bioimaging.
- To investigate the use of aromatic alkenes, specifically styrene, as a dipolarophile in this reaction.
Main Methods:
- Investigated styrene as a dipolarophile in the alkene-tetrazole cycloaddition reaction.
- Performed mechanistic studies to understand the reaction improvements.
- Demonstrated fluorogenic protein labeling in vitro and in vivo using genetically incorporated unnatural amino acids.
Main Results:
- Achieved over a 30-fold improvement in the quantum yield of the reaction product in aqueous solution.
- Identified potential mechanistic reasons for the improvement, related to protonation of the reaction product.
- Successfully demonstrated fluorogenic protein labeling in vitro and in vivo.
Conclusions:
- The styrene-tetrazole reaction significantly enhances fluorophore quantum yield, offering a valuable tool for bioorthogonal chemistry.
- Findings provide guidance for designing improved alkene-tetrazole reactions for biological studies.
- The strategy is applicable for real-time imaging in live cells and can be expanded with various tetrazole derivatives.