1. Hydrophilic trans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling.
- Author
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Kozma E, Nikić I, Varga BR, Aramburu IV, Kang JH, Fackler OT, Lemke EA, and Kele P
- Subjects
- Animals, COS Cells, Cells, Cultured, Chlorocebus aethiops, HEK293 Cells, Humans, Hydrophobic and Hydrophilic Interactions, Microscopy, Fluorescence, Molecular Structure, Amino Acids chemistry, Cyclooctanes chemistry, Fluorescent Dyes chemistry, Proteins chemistry
- Abstract
Introduction of bioorthogonal functionalities (e.g., trans-cyclooctene-TCO) into a protein of interest by site-specific genetic encoding of non-canonical amino acids (ncAAs) creates uniquely targetable platforms for fluorescent labeling schemes in combination with tetrazine-functionalized dyes. However, fluorescent labeling of an intracellular protein is usually compromised by high background, arising from the hydrophobicity of ncAAs; this is typically compensated for by hours-long washout to remove excess ncAAs from the cellular interior. To overcome these problems, we designed, synthesized, and tested new, hydrophilic TCO-ncAAs. One derivative, DOTCO-lysine was genetically incorporated into proteins with good yield. The increased hydrophilicity shortened the excess ncAA washout time from hours to minutes, thus permitting rapid labeling and subsequent fluorescence microscopy., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2016
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