Protein interactor detection systems
OwnerXiaokun SHU, William Degrado +2inventors
InventorsXiaokun SHU, William Degrado, Tsz-Leung To, Hyunil Jo
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Abstract
Provided herein are systems, methods and reagents for determining interactors (proteins or nucleic acids) that interact with a protein of interest. The subject system, methods and reagents advantageously allow for the identification of weak and transient protein-protein interactions. Such subject system, methods and reagents are useful, for example, for the determination of specific protein-interactor interactions that exist in particular diseases. Determination of these differences is useful, for example, in the drug development for the treatment of such diseases.
Claims6
-20. (canceled)
A system for determining interactors that interact with a protein of interest comprising:
- a SOG-POI protein comprising a singlet oxygen photosensitizer fused to a protein of interest, wherein the singlet oxygen photosensitizer is capable of producing singlet oxygen (1O2) when illuminated with a light source;
- and an interactor detection molecule according to the formula:
- wherein:
- X is a label;
- A is an optional linker wherein the linker A is a C1-C10 alkylene, an aryl, or a heteroaryl;
- and
- Y is a reactive group capable of interaction with a cysteine, lysine, histidine, or serine side chain upon oxidation by a singlet oxygen (1O2).
A cell comprising:
- a SOG-POI protein comprising a singlet oxygen photosensitizer fused to a protein of interest, wherein the singlet oxygen photosensitizer is capable of producing singlet oxygen (1O2) when illuminated with a light source;
- and
- an interactor detection molecule according to the formula:
- wherein:
- X is a label;
- A is an optional linker wherein the linker A is a C1-C10 alkylene, an aryl, or a heteroaryl;
- and
- Y is a reactive group capable of interaction with a cysteine, lysine, histidine, or serine side chain upon oxidation by a singlet oxygen (1O2).
A method for determining interactors that interact with a protein of interest comprising:
- a) introducing into a cell a SOG-POI protein and an interactor detection molecule according to the formula:
- wherein:
- X is a label;
- A is an optional linker wherein the linker A is a C1-C10 alkylene, an aryl, or a heteroaryl;
- and
- Y is a reactive group capable of interaction with a cysteine, lysine, histidine, or serine side chain upon oxidation by a singlet oxygen (1O2),
- wherein the SOG-POI protein comprises a singlet oxygen photosensitizer fused to a protein of interest, and wherein the singlet oxygen photosensitizer is capable of producing singlet oxygen (1O2) when illuminated with a light source;
- b) illuminating the singlet oxygen photosensitizer with a light source, thereby producing singlet oxygen (1O2) that oxidizes the interactor detection molecule to form a reactive detection intermediate, wherein the reactive detection intermediate binds with a interaction protein that interacts with the protein of interest, thereby labeling the interactor;
- and
- c) characterizing the interactor.
Dependencies are inferred from the claim text.