Hydrophilictrans-Cyclooctenylated Noncanonical Amino Acids for Fast Intracellular Protein Labeling
| Autor: | Ivana Nikić, Péter Kele, Iker Valle Aramburu, Eszter Kozma, Jun Hee Kang, Balázs Varga, Edward A. Lemke, Oliver T. Fackler |
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| Rok vydání: | 2016 |
| Předmět: |
education
010402 general chemistry 01 natural sciences Biochemistry Cyclooctanes Chlorocebus aethiops Fluorescence microscope Animals Humans Amino Acids Molecular Biology Cells Cultured Fluorescent Dyes chemistry.chemical_classification COS cells Molecular Structure 010405 organic chemistry Organic Chemistry HEK 293 cells Proteins Protein engineering Fluorescence 0104 chemical sciences Amino acid HEK293 Cells Microscopy Fluorescence chemistry COS Cells Click chemistry Molecular Medicine Bioorthogonal chemistry Hydrophobic and Hydrophilic Interactions human activities |
| Zdroj: | ChemBioChem. 17:1518-1524 |
| ISSN: | 1439-4227 |
| DOI: | 10.1002/cbic.201600284 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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