Method for detecting protein-protein interaction
View Patent ↗A method for detecting an interaction between a first protein and a second protein comprises the steps of: expressing in a cell a first fusion protein comprising the first protein and an association-inducing protein, and a second fusion protein comprising the second protein and a fluorescent protein having a multimerization ability; detecting a fluorescent focus formed by an association between the first fusion protein and the second fusion protein in the cell; and determining an interaction between the first protein and the second protein according to the detection of the fluorescent focus.
1. A method for screening for a substance modulating an interaction between a first protein and a second protein, the method comprising the steps of:
expressing in a cell a first fusion protein comprising the first protein and an association-inducing protein, and a second fusion protein comprising the second protein and a fluorescent protein having a multimerization ability and emitting fluorescence, in presence of a test compound;
detecting a fluorescent focus formed by an association between the first fusion protein and the second fusion protein in the cell; and
selecting the test compound as a substance inducing the interaction if a fluorescence intensity of the fluorescent focus is higher than a fluorescence intensity of a fluorescent focus formed in absence of the test compound, or selecting the test compound as a substance suppressing the interaction if the fluorescence intensity of the fluorescent focus is lower than the fluorescence intensity of the fluorescent focus formed in the absence of the test compound, wherein
the association-inducing protein is at least one protein selected from the group consisting of a PB1 domain of p62, a PB1 domain of TFG, a PB1 domain of PKCiota, a SAM domain of TEL, a SAM domain of DGK delta, and a SAM domain of Tankyrase-1;
the fluorescent protein having a multimerization ability is at least one fluorescent protein selected from the group consisting of a florescent protein capable of forming a homomultimer, monomeric Kusabira-Orange 2, monomeric Keima-Red, monomeric Midoriishi-Cyan1, monomeric Kusabira-Orange 1, and monomeric Kikume Green-Red1;
the fluorescent focus has a fluorescence intensity in a region of 0.2 to 5 μm, the fluorescence intensity being higher than a fluorescence intensity of the fluorescent protein having a multimerization ability, which is present in a dispersed state in the cell, and
the association inducing protein is able to form a fluorescent focus when fused to the fluorescent protein having a multimerization ability.