Method and antibodies for the identification of ubiquitinated proteins and sites of ubiquitination
AssigneeSyddansk Universitet
InventorsBlagoy BLAGOEV, Jens VANSELOW, Vyacheslav AKIMOV, Mogens NIELSEN
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Abstract
There is provided antibodies, epitopes and methods for detecting ubiquitinated polypeptides and ubiquitination sites in proteins. The antibodies recognize a fragment of ubiquitin that is created after samples are treated with the proteoloytical enzyme LysC (or LysN).
Claims11
An antibody that specifically binds to an epitope having the sequence ESTLHLVLRLRGG (SEQ ID NO 1), said antibody being able to specifically distinguish between ubiquitin and ubiquitin-like modified peptides.
An isolated epitope having the sequence ESTLHLVLRLRGG (SEQ ID NO 1).
A method of detecting a ubiquitinated site in a test protein or a mixture of test proteins that comprises
- (i) cleaving a test protein or a mixture of test proteins with LysC that cleaves ubiquitin to form a mixture of cleavage peptides;
- (ii) contacting the cleavage peptides with an antibody that specifically binds to an epitope having the sequence ESTLHLVLRLRGG (SEQ ID NO 1), said antibody being able to specifically distinguish between ubiquitin and ubiquitin-like modified peptides;
- and
- (iii) analysing the peptide(s) that bind to the antibody thereby detecting and identifying a ubiquitinated site in a test protein or a mixture of test proteins.
Dependencies are inferred from the claim text.