Methods and compositions for protein labeling using lipoic acid ligases
AssigneesMassachusetts Institute of Technology, University of Washington through its Center for Commercialization
InventorsAlice Y. Ting, David Baker, Florian Richter, Lucas Nivon, Daniel Shao-Chen Liu
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Abstract
The present disclosure 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 (e.g., lipoic acid analogs comprising a resorufin moiety) recognized by lipoic acid ligase and lipoic acid ligase mutants. Also provided herein is a method of imaging protein-protein interaction via a reaction mediated by lipoic acid ligase.
Claims36
A lipoic acid ligase mutant that is specific to resorufin, the mutant comprising an amino acid sequence that is at least 85% identical to SEQ ID NO:11 and includes mutations at residues corresponding to E20 and F147 in SEQ ID NO:11.
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9-20. (canceled)
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34-37. (canceled)
The method of claim 37, wherein the acceptor polypeptide comprises the amino acid sequence GFEIDKVWYDLDA (SEQ ID NO:32).
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44-47. (canceled)
The kit of claim 47, wherein the acceptor polypeptide comprises the amino acid sequence GFEIDKVWYDLDA (SEQ ID NO:32).
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An isolated lipoic acid analog compound, which is (i) a compound comprising an aliphatic carboxylic acid substituted with a resorufin moiety, or (ii) resorufin-AM2.
Dependencies are inferred from the claim text.