Virus-like particle (VLP) based small molecule-protein interaction trap
AssigneesUniversiteit Gent, Vib Vzw
InventorsJan Tavernier, Sven Eyckerman
Life of this patent
Estimating the term…
Abstract
This disclosure relates to a virus-like particle in which a small molecule-protein complex is entrapped, ensuring the formation of the small molecule-protein complex under physiological conditions, while protecting the small molecule-protein complex during purification and identification. The disclosure further relates to the use of such virus-like particle for the isolation and identification of small molecule-protein complexes.
Claims19
A method of detecting a small molecule-protein interaction in a cell, the method comprising:
- isolating an artificial VLP formed by the cell;
- detecting a small-molecule-protein complex entrapped in the artificial VLP;
- wherein the small-molecule-protein complex entrapped in the artificial VLP comprises:
- a VLP-forming polypeptide or a fusion protein comprising a VLP-forming polypeptide;
- a fusion construct comprising first and second small molecules covalently linked to one another, wherein the first small molecule interacts with the VLP-forming polypeptide or fusion protein and the second small molecule interacts with at least one polypeptide other than the VLP-forming polypeptide or fusion protein;
- and
- a polypeptide other than the VLP-forming polypeptide or fusion protein that interacts with the second small molecule.
A method for detecting a small molecule-protein interaction, the method comprising:
- expressing a virus-like particle (VLP)-forming polypeptide or a fusion protein comprising a VLP-forming polypeptide in a cell;
- recruiting a fusion construct comprising first and second small molecules covalently linked to one another, wherein the first small molecule interacts with the VLP-forming polypeptide or the fusion protein and the second small molecule interacts with at least one polypeptide other than the VLP-forming polypeptide or fusion protein, to said VLP-forming polypeptide;
- interacting a polypeptide other than the VLP-forming polypeptide or fusion protein with the second small molecule to the fusion construct;
- isolating the VLPs;
- and
- analyzing an entrapped complex in an isolated VLP.
A method of detecting a small molecule-protein interaction in a cell, the method comprising:
- isolating an artificial VLP formed by the cell;
- detecting a small-molecule-protein complex entrapped in the artificial VLP;
- wherein the small-molecule-protein complex entrapped in the artificial VLP comprises:
- a VLP-forming polypeptide or a fusion protein comprising a VLP-forming polypeptide;
- a fusion construct comprising first and second small molecules covalently linked to one another, wherein the first small molecule interacts with the VLP-forming polypeptide or fusion protein and the second small molecule interacts with at least one wild-type polypeptide;
- and
- a wild-type polypeptide that interacts with the second small molecule.
Dependencies are inferred from the claim text.