Methods and kits for detecting protein-protein interactions

The present invention relates to methods and kits for detection protein-protein interactions. In particular, the present invention relates to a method for detecting the binding between a first polypeptide (A) and a second polypeptide (B) in a cell comprising: i) providing a cell that expresses: (a) a polypeptide (GFP1-9) comprising an amino acid sequence having at least 90% of identity with the amino acid sequence selected from the group consisting of SEQ ID NO:1-4 (b) a first fusion protein wherein the polypeptide (A) is fused to a polypeptide (GFP10) having an amino an amino acid sequence having at least 90% of identity with the amino acid sequence selected from the group consisting of SEQ ID NO:5-7 (c) a second fusion protein wherein the polypeptide (B) is fused to a polypeptide (GFP11) having an amino an amino acid sequence having at least 90% of identity with the amino acid sequence selected from the group consisting of SEQ ID NO:8-9 and (d) an intrabody specific for the complex formed by the self-assembly of the first, second and third polypeptides (a), (b) and (c) ii) detecting the fluorescence wherein when the fluorescence is detected it is concluded that the polypeptide (A) binds to polypeptide (B) and wherein the fluorescence is not detected it is concluded that the polypeptides (A) does not bind to polypeptide (B).

Keywords: protein-protein interaction, screening, GTPase, split-GFP
Patent Application number: EP 15 305 971.2
Inventors:
Stéphanie CABANTOUSFaten KORAÏCHIGilles FAVRE
Publications:
Sci Rep. 2013 Oct 4;3:2854. doi: 10.1038/srep02854.

Reference:

BIO15012-R1

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Patent filling date: 24-06-2015

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