Compositions and methods for detecting molecule-molecule interactions
AssigneeShanghaitech University
InventorsMin Zhuang, Haopeng Wang, Qiang Liu
Life of this patent
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Abstract
Compositions and methods for detecting molecule-molecule interactions are provided. The methods employ a prokaryotic ubiquitin-like protein (Pup) and a Pup ligase that is coupled to one of the molecules. When the Pup ligase is brought to proximity to the other molecule by virtue of the molecule-molecule interaction, the Pup ligase can conjugate the Pup to a lysine residue on the other molecule. As such conjugation can be easily detected, this method allows easy identification of the molecule-molecule interaction.
Claims19
A method of detecting binding between a protein having at least a lysine residue and a molecule, comprising:
- incubating a sample that comprises (a) a prokaryotic ubiquitin-like protein (Pup), (b) the protein, and (c) the molecule coupled to a Pup ligase, under conditions to allow the protein to bind the molecule, thereby allowing the Pup ligase to conjugate the Pup to the lysine residue of the protein;
- and
- detecting the conjugation of the Pup to the protein which indicates binding between the protein and the molecule.
A method of coupling a molecule to a protein having at least a lysine residue, comprising contacting the protein with (a) the molecule coupled to a prokaryotic ubiquitin-like protein (Pup) in the presence of (b) a Pup ligase coupled to a second protein capable of binding the protein, under conditions to allow the second protein to bind the protein thereby enabling the Pup ligase to conjugate the Pup to the lysine residue, thereby coupling the molecule to the protein.
(canceled)
A complex comprising a prokaryotic ubiquitin-like protein (Pup) ligase coupled to a transmembrane protein.
19-21. (canceled)
Dependencies are inferred from the claim text.