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 journalArticle

113 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)2094-2099
Number of pages6
JournalChemBioChem
Volume13
Issue number14
DOIs
StatePublished - Sep 24 2012
Externally publishedYes

Fingerprint

Ligation
Metals
Imaging techniques
Amino Acids
Cycloaddition
Cycloaddition Reaction
Proteins
Coloring Agents
Alkynes
Azides
Biomolecules
Labeling
Cells
bicyclo(6.1.0)nonyne
Electrons
Kinetics

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

Cite this

Borrmann, A., Milles, S., Plass, T., Dommerholt, J., Verkade, J. M. M., Wießler, M., ... Lemke, E. A. (2012). Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation. ChemBioChem, 13(14), 2094-2099. https://doi.org/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. / Borrmann, Annika; Milles, Sigrid; Plass, Tilman; Dommerholt, Jan; Verkade, Jorge M M; Wießler, Manfred; Schultz, Carsten; van Hest, Jan C M; van Delft, Floris L.; Lemke, Edward A.

In: ChemBioChem, Vol. 13, No. 14, 24.09.2012, p. 2094-2099.

Research output: Contribution to journalArticle

Borrmann, A, Milles, S, Plass, T, Dommerholt, J, Verkade, JMM, Wießler, M, Schultz, C, van Hest, JCM, van Delft, FL & Lemke, EA 2012, 'Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation', ChemBioChem, vol. 13, no. 14, pp. 2094-2099. https://doi.org/10.1002/cbic.201200407
Borrmann, Annika ; Milles, Sigrid ; Plass, Tilman ; Dommerholt, Jan ; Verkade, Jorge M M ; Wießler, Manfred ; Schultz, Carsten ; van Hest, Jan C M ; van Delft, Floris L. ; Lemke, Edward A. / Genetic Encoding of a Bicyclo[6.1.0]nonyne-Charged Amino Acid Enables Fast Cellular Protein Imaging by Metal-Free Ligation. In: ChemBioChem. 2012 ; Vol. 13, No. 14. pp. 2094-2099.
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