Chapter title |
Tub-Tag Labeling; Chemoenzymatic Incorporation of Unnatural Amino Acids
|
---|---|
Chapter number | 4 |
Book title |
Noncanonical Amino Acids
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7574-7_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7573-0, 978-1-4939-7574-7
|
Authors |
Jonas Helma, Heinrich Leonhardt, Christian P. R. Hackenberger, Dominik Schumacher |
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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