Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation

Annika Borrmann; Sigrid Milles; Tilman Plass; Jan Dommerholt; Jorge M.M. Verkade; Manfred Wießler; Carsten Schultz; Jan C.M. van Hest; Floris L. van Delft; Edward A. Lemke

doi:10.1002/cbic.201200407

Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation

Annika Borrmann, Sigrid Milles, Tilman Plass, Jan Dommerholt, Jorge M.M. Verkade, Manfred Wießler, Carsten Schultz, Jan C.M. van Hest, Floris L. van Delft, Edward A. Lemke

Research output: Contribution to journal › Article › peer-review

142 Scopus citations

Abstract

Visualizing biomolecules by fluorescent tagging is a powerful method for studying their behaviour and function inside cells. We prepared and genetically encoded an unnatural amino acid (UAA) that features a bicyclononyne moiety. This UAA offered exceptional reactivity in strain-promoted azide-alkyne cycloadditions. Kinetic measurements revealed that the UAA reacted also remarkably fast in the inverse-electron-demand Diels-Alder cycloaddition with tetrazine-conjugated dyes. Genetic encoding of the new UAA inside mammalian cells and its subsequent selective labeling at low dye concentrations demonstrate the usefulness of the new amino acid for future imaging studies.

Original language	English (US)
Pages (from-to)	2094-2099
Number of pages	6
Journal	ChemBioChem
Volume	13
Issue number	14
DOIs	https://doi.org/10.1002/cbic.201200407
State	Published - Sep 24 2012
Externally published	Yes

Keywords

Amber suppression
Click chemistry
Diels-Alder reaction
Protein engineering
Unnatural amino acid

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Organic Chemistry

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10.1002/cbic.201200407

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title = "Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation",

abstract = "Visualizing biomolecules by fluorescent tagging is a powerful method for studying their behaviour and function inside cells. We prepared and genetically encoded an unnatural amino acid (UAA) that features a bicyclononyne moiety. This UAA offered exceptional reactivity in strain-promoted azide-alkyne cycloadditions. Kinetic measurements revealed that the UAA reacted also remarkably fast in the inverse-electron-demand Diels-Alder cycloaddition with tetrazine-conjugated dyes. Genetic encoding of the new UAA inside mammalian cells and its subsequent selective labeling at low dye concentrations demonstrate the usefulness of the new amino acid for future imaging studies.",

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AU - Milles, Sigrid

AU - Plass, Tilman

AU - Dommerholt, Jan

AU - Verkade, Jorge M.M.

AU - Wießler, Manfred

AU - Schultz, Carsten

AU - van Hest, Jan C.M.

AU - van Delft, Floris L.

AU - Lemke, Edward A.

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AB - Visualizing biomolecules by fluorescent tagging is a powerful method for studying their behaviour and function inside cells. We prepared and genetically encoded an unnatural amino acid (UAA) that features a bicyclononyne moiety. This UAA offered exceptional reactivity in strain-promoted azide-alkyne cycloadditions. Kinetic measurements revealed that the UAA reacted also remarkably fast in the inverse-electron-demand Diels-Alder cycloaddition with tetrazine-conjugated dyes. Genetic encoding of the new UAA inside mammalian cells and its subsequent selective labeling at low dye concentrations demonstrate the usefulness of the new amino acid for future imaging studies.

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KW - Click chemistry

KW - Diels-Alder reaction

KW - Protein engineering

KW - Unnatural amino acid

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Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation

Abstract

Keywords

ASJC Scopus subject areas

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