Minimal tags for rapid dual-color live-cell labeling and super-resolution microscopy
Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim..
The growing demands of advanced fluorescence and super-resolution microscopy benefit from the development of small and highly photostable fluorescent probes. Techniques developed to expand the genetic code permit the residue-specific encoding of unnatural amino acids (UAAs) armed with novel clickable chemical handles into proteins in living cells. Here we present the design of new UAAs bearing strained alkene side chains that have improved biocompatibility and stability for the attachment of tetrazine-functionalized organic dyes by the inverse-electron-demand Diels-Alder cycloaddition (SPIEDAC). Furthermore, we fine-tuned the SPIEDAC click reaction to obtain an orthogonal variant for rapid protein labeling which we termed selectivity enhanced (se) SPIEDAC. seSPIEDAC and SPIEDAC were combined for the rapid labeling of live mammalian cells with two different fluorescent probes. We demonstrate the strength of our method by visualizing insulin receptors (IRs) and virus-like particles (VLPs) with dual-color super-resolution microscopy.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2014 |
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| Erschienen: |
2014 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:53 |
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| Enthalten in: |
Angewandte Chemie (International ed. in English) - 53(2014), 8 vom: 17. Feb., Seite 2245-9 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Nikić, Ivana [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 01.12.2014 Date Revised 09.05.2014 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1002/anie.201309847 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM234946237 |
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| 245 | 1 | 0 | |a Minimal tags for rapid dual-color live-cell labeling and super-resolution microscopy |
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| 500 | |a Date Revised 09.05.2014 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. | ||
| 520 | |a The growing demands of advanced fluorescence and super-resolution microscopy benefit from the development of small and highly photostable fluorescent probes. Techniques developed to expand the genetic code permit the residue-specific encoding of unnatural amino acids (UAAs) armed with novel clickable chemical handles into proteins in living cells. Here we present the design of new UAAs bearing strained alkene side chains that have improved biocompatibility and stability for the attachment of tetrazine-functionalized organic dyes by the inverse-electron-demand Diels-Alder cycloaddition (SPIEDAC). Furthermore, we fine-tuned the SPIEDAC click reaction to obtain an orthogonal variant for rapid protein labeling which we termed selectivity enhanced (se) SPIEDAC. seSPIEDAC and SPIEDAC were combined for the rapid labeling of live mammalian cells with two different fluorescent probes. We demonstrate the strength of our method by visualizing insulin receptors (IRs) and virus-like particles (VLPs) with dual-color super-resolution microscopy | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a amino acids | |
| 650 | 4 | |a click chemistry | |
| 650 | 4 | |a cycloaddition | |
| 650 | 4 | |a protein engineering | |
| 650 | 4 | |a super-resolution microscopy | |
| 650 | 7 | |a Amino Acids |2 NLM | |
| 650 | 7 | |a Fluorescent Dyes |2 NLM | |
| 650 | 7 | |a Hemagglutinins |2 NLM | |
| 650 | 7 | |a Viral Matrix Proteins |2 NLM | |
| 650 | 7 | |a Viral Proteins |2 NLM | |
| 650 | 7 | |a Receptor, Insulin |2 NLM | |
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| 700 | 1 | |a Plass, Tilman |e verfasserin |4 aut | |
| 700 | 1 | |a Schraidt, Oliver |e verfasserin |4 aut | |
| 700 | 1 | |a Szymański, Jędrzej |e verfasserin |4 aut | |
| 700 | 1 | |a Briggs, John A G |e verfasserin |4 aut | |
| 700 | 1 | |a Schultz, Carsten |e verfasserin |4 aut | |
| 700 | 1 | |a Lemke, Edward A |e verfasserin |4 aut | |
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