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US 8,137,925 B2Granted
Methods and compositions for protein labeling using lipoic acid ligases
AssigneeMassachusetts Institute of Technology
InventorsAlice Y. Ting, Marta Fernandez Suarez, Hemanta Baruah
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Abstract
The invention provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs recognized by lipoic acid ligase and lipoic acid ligase mutants.
Claims20
A method for labeling a target protein, the method comprising
- contacting a fusion protein with a lipoic acid analog, and
- allowing sufficient time for the lipoic acid analog to be conjugated to the fusion protein via an acceptor polypeptide, in the presence of a lipoic acid ligase polypeptide that comprises an amino acid sequence of a wild-type lipoic acid ligase or a mutant thereof, the mutant being at least 90% identical to SEQ ID NO:11 and having lipoic acid ligase activity;
- wherein the fusion protein is a fusion of the target protein and the acceptor polypeptide, which comprises an amino acid sequence at least 90% identical to SEQ ID NO:
- 3, 5, 6, 8, or 10, and
- wherein the lipoic acid analog comprises an alkyl azide, an alkyne, an alkyl halide, an aryl azide photoaffinity probe, a diazirine photoaffinity probe, a benzophenone photoaffinity probe, or a fluorophore.
2 more elements
Dependencies are inferred from the claim text.