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Methods Mol Biol. 2018;1728:67-93. doi: 10.1007/978-1-4939-7574-7_4.

Tub-Tag Labeling; Chemoenzymatic Incorporation of Unnatural Amino Acids.

Author information

1
Department of Biology II, Center for Integrated Protein Science Munich, Ludwig Maximilians Universität München, Planegg-Martinsried, Germany.
2
Department of Chemical-Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Berlin, Germany.
3
Department of Chemistry, Humboldt Universität zu Berlin, Berlin, Germany.
4
Department of Chemical-Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Berlin, Germany. dschumacher@fmp-berlin.de.
5
Department of Chemistry, Humboldt Universität zu Berlin, Berlin, Germany. dschumacher@fmp-berlin.de.

Abstract

Tub-tag labeling is a chemoenzymatic method that enables the site-specific labeling of proteins. Here, the natural enzyme tubulin tyrosine ligase incorporates noncanonical tyrosine derivatives to the terminal carboxylic acid of proteins containing a 14-amino acid recognition sequence called Tub-tag. The tyrosine derivative carries a unique chemical reporter allowing for a subsequent bioorthogonal modification of proteins with a great variety of probes. Here, we describe the Tub-tag protein modification protocol in detail and explain its utilization to generate labeled proteins for advanced applications in cell biology, imaging, and diagnostics.

KEYWORDS:

Bioorthogonal reaction; Chemoenzymatic labeling; Site-specific protein functionalization; Tubulin tyrosine ligase; Tyrosine derivatives

PMID:
29404991
DOI:
10.1007/978-1-4939-7574-7_4
[Indexed for MEDLINE]

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