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Versatile and Efficient Site‐Specific Protein Functionalization by Tubulin Tyrosine Ligase
Author(s) -
Schumacher Dominik,
Helma Jonas,
Mann Florian A.,
Pichler Garwin,
Natale Francesco,
Krause Eberhard,
Cardoso M. Cristina,
Hackenberger Christian P. R.,
Leonhardt Heinrich
Publication year - 2015
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.201505456
Subject(s) - tyrosine , dna ligase , biochemistry , tubulin , bioorthogonal chemistry , ubiquitin ligase , biology , chemistry , protein engineering , microbiology and biotechnology , ubiquitin , enzyme , combinatorial chemistry , microtubule , click chemistry , gene
A novel chemoenzymatic approach for simple and fast site‐specific protein labeling is reported. Recombinant tubulin tyrosine ligase (TTL) was repurposed to attach various unnatural tyrosine derivatives as small bioorthogonal handles to proteins containing a short tubulin‐derived recognition sequence (Tub‐tag). This novel strategy enables a broad range of high‐yielding and fast chemoselective C‐terminal protein modifications on isolated proteins or in cell lysates for applications in biochemistry, cell biology, and beyond, as demonstrated by the site‐specific labeling of nanobodies, GFP, and ubiquitin.