Premium
Covalent Protein Labeling by SpyTag–SpyCatcher in Fixed Cells for Super‐Resolution Microscopy
Author(s) -
Pessino Veronica,
Citron Y. Rose,
Feng Siyu,
Huang Bo
Publication year - 2017
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201700177
Subject(s) - epitope , gene isoform , protein tag , chemistry , covalent bond , fluorescence microscope , super resolution microscopy , fusion protein , resolution (logic) , intracellular , target protein , biochemistry , fluorescence , biophysics , microbiology and biotechnology , biology , antibody , gene , recombinant dna , genetics , computer science , physics , organic chemistry , quantum mechanics , artificial intelligence
Labeling proteins with high specificity and efficiency is a fundamental prerequisite for microscopic visualization of subcellular protein structures and interactions. Although the comparatively small size of epitope tags makes them less perturbative to fusion proteins, they require the use of large antibodies that often limit probe accessibility and effective resolution. Here we use the covalent SpyTag–SpyCatcher system as an epitope‐like tag for fluorescent labeling of intracellular proteins in fixed cells for both conventional and super‐resolution microscopy. We also applied this method to endogenous proteins by gene editing, demonstrating its high labeling efficiency and capability for isoform‐specific labeling.