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Published on: December 19, 2020
Tub-Tag Labeling; Chemoenzymatic Incorporation of Unnatural Amino Acids
Jonas Helma1, Heinrich Leonhardt1, Christian P R Hackenberger2,3
1Department of Biology II, Center for Integrated Protein Science Munich, Ludwig Maximilians Universität München, Planegg-Martinsried, Germany.
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.
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