Minimal tags for rapid dual-color live-cell labeling and super-resolution microscopy

Ivana Nikic̈, Tilman Plass, Oliver Schraidt, Jȩdrzej Szymański, John A.G. Briggs, Carsten Schultz, Edward A. Lemke

Research output: Contribution to journalArticle

168 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2245-2249
Number of pages5
JournalAngewandte Chemie - International Edition
Volume53
Issue number8
DOIs
StatePublished - Feb 17 2014

Keywords

  • amino acids
  • click chemistry
  • cycloaddition
  • protein engineering
  • super-resolution microscopy

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)

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