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Angew Chem Int Ed Engl. 2016 Nov 2;55(45):14046-14050. doi: 10.1002/anie.201608009. Epub 2016 Oct 13.

Optimized Tetrazine Derivatives for Rapid Bioorthogonal Decaging in Living Cells.

Author information

1
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.
2
Peking-Tsinghua Center for Life Sciences, Beijing, 100871, China.
3
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China. pengchen@pku.edu.cn.
4
Peking-Tsinghua Center for Life Sciences, Beijing, 100871, China. pengchen@pku.edu.cn.

Abstract

The inverse-electron-demand Diels-Alder (iDA) reaction has recently been repurposed as a bioorthogonal decaging reaction by accelerating the elimination process after an initial cycloaddition between trans-cyclooctene (TCO) and tetrazine (TZ). Herein, we systematically surveyed 3,6-substituted TZ derivatives by using a fluorogenic TCO-coumarin reporter followed by LC-MS analysis, which revealed that the initial iDA cycloaddition step was greatly accelerated by electron-withdrawing groups (EWGs) while the subsequent elimination step was strongly suppressed by EWGs. In addition, smaller substituents facilitated the decaging process. These findings promoted us to design and test unsymmetric TZs bearing an EWG group and a small non-EWG group at the 3- and 6-position, respectively. These TZs showed remarkably enhanced decaging rates, enabling rapid iDA-mediated protein activation in living cells.

KEYWORDS:

Diels-Alder reactions; amino acids; decaging; protein activation; protein engineering

PMID:
27735133
DOI:
10.1002/anie.201608009
[Indexed for MEDLINE]

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