Selective modulation of protein-protein interactions
AssigneeSyntheX, Inc.
InventorsMaria Soloveychik, Charly Chahwan
Life of this patent
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Abstract
The present disclosure provides methods to identify peptides and small molecule moieties that are able to modulate protein-protein interactions (PPIs). Some moieties can disrupt specific PPIs within a complex, or disrupt variant-specific PPIs. Some moieties can alternatively bridge between two proteins in a protein-specific or a variant-specific manner. The methods described enable generation of compounds able to modulate PPI networks within cells with implications for drug development for pathological conditions.
Claims31
A method for identifying a molecule that disrupts an interaction between a first test protein and a second test protein in a host cell, the method comprising:
- expressing in the host cell a first fusion protein comprising the first test protein and a first DNA-binding moiety;
- expressing in the host cell a second fusion protein comprising the second test protein and a gene activating moiety;
- expressing in the host cell a third fusion protein comprising a third test protein and a second DNA-binding moiety, wherein the second DNA-binding moiety is different from the first DNA-binding moiety;
- and
- delivering a molecule from a library to the host cell,
- wherein a sequence of a gene for expressing a death agent is disposed within the host cell and operably linked to a promoter DNA sequence specific for the first DNA-binding moiety,
- wherein a positive selection reporter is disposed within the host cell and operably linked to a promoter DNA sequence specific for the second DNA binding moiety, and
- wherein, in an absence of the molecule, an interaction between the first test protein and the second test protein causes the gene activating moiety to activate expression of the death agent, while an interaction between the second test protein and the third test protein causes the gene activating moiety to activate expression of the positive selection reporter.
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A host cell configured to express:
- a first fusion protein comprising a first DNA-binding moiety;
- a second fusion protein comprising a gene activating moiety;
- a third fusion protein comprising a second DNA-binding moiety, wherein the second DNA-binding moiety is different from the first DNA-binding moiety;
- a death agent, wherein expression of the death agent is under control of a promoter DNA sequence specific for the first DNA-binding moiety;
- a positive selection reporter, wherein expression of the positive selection reporter is under control of a promoter DNA sequence specific for the second DNA-binding moiety;
- and
- a polypeptide of 60 or fewer amino acids, wherein the polypeptide modulates an interaction between the first test protein and the second test protein,
- wherein the host cell optionally has a mutant background enabling uptake of or reducing efflux of small molecules, and
- wherein the host cell optionally has a mutant background enabling increased transformation efficiency.
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A method for identifying a molecule that selectively facilitates an interaction between a first test protein and a second test protein, the method comprising:
- expressing in a host cell a first fusion protein comprising the first test protein and a first DNA-binding moiety;
- expressing in the host cell a second fusion protein comprising the second test protein and a gene activating moiety;
- expressing in the host cell a third fusion protein comprising a third test protein and a second DNA-binding moiety, wherein the second DNA-binding moiety is different from the first DNA-binding moiety;
- and
- delivering a molecule from a library to the host cell such that the molecule forms a bridging interaction between the first test protein and the second test protein,
- wherein a sequence of a gene for expressing a death agent is disposed within the host cell and operably linked to a promoter DNA sequence specific for the second DNA-binding moiety,
- wherein a positive selection reporter is disposed within the host cell and operably linked to a promoter DNA sequence specific for the first DNA binding moiety, and
- wherein the first test protein and the second test protein form a functional transcription factor that activates expression of the positive selection reporter when the molecule from the library forms the bridging interaction.
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Dependencies are inferred from the claim text.