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US 10,208,084 B2Granted
Means and methods for site-specific functionalization of polypeptides
AssigneesLudwig-Maximilians-Universitat Munchen, Forschungsverbund Berlin E.V.
InventorsHeinrich Leonhardt, Jonas Helma, Dominik Schumacher, Christian Hackenberger
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Abstract
The present invention provides means and methods for equipping a polypeptide of interest at its C-terminus with a versatile adaptor amino acid that allows the functionalization of the polypeptide of interest.
Claims31
A method for the production of a polypeptide comprising
- (a) introducing or adding at the C-terminus of a polypeptide a recognition sequence for tubulin tyrosine ligase;
- and
- (b) contacting the polypeptide obtained in step (a) in the presence of tubulin tyrosine ligase and a tyrosine derivative under conditions suitable for the tubulin tyrosine ligase to tyrosinate said polypeptide with said tyrosine derivative.
A method for the production of a polypeptide comprising
- (a′) introducing or adding at the C-terminus of a polypeptide a recognition sequence for tubulin tyrosine ligase;
- and
- (b′) contacting the polypeptide obtained in step (a′) in the presence of tubulin tyrosine ligase and a tyrosine derivative conjugated to a moiety under conditions suitable for the tubulin tyrosine ligase to tyrosinate said polypeptide with said tyrosine derivative conjugated to said moiety.
A method for installing a click chemistry handle to the C-terminus of a polypeptide other than tubulin, comprising:
- (a) providing a polypeptide having at its C-terminus a tubulin tyrosine ligase recognition sequence;
- and
- (b) contacting the polypeptide of step (a) in the presence of tubulin tyrosine ligase and a tyrosine derivative containing an unnatural functional group for chemoselective or bioorthogonal modifications under conditions suitable for the tubulin tyrosine ligase to tyrosinate said polypeptide with said tyrosine derivative.
Dependencies are inferred from the claim text.