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Minimal Tags for Rapid Dual‐Color Live‐Cell Labeling and Super‐Resolution Microscopy
Author(s) -
Nikić Ivana,
Plass Tilman,
Schraidt Oliver,
Szymański Jędrzej,
Briggs John A. G.,
Schultz Carsten,
Lemke Edward A.
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201309847
Subject(s) - tetrazine , fluorescence microscope , cycloaddition , bioorthogonal chemistry , fluorescence , protein tag , click chemistry , super resolution microscopy , chemistry , genetic code , microscopy , live cell imaging , amino acid , biophysics , nanotechnology , combinatorial chemistry , biochemistry , materials science , cell , biology , fusion protein , physics , organic chemistry , quantum mechanics , optics , gene , recombinant dna , catalysis
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.