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1. (WO2014197986) DETECTION OF PROTEIN TO PROTEIN INTERACTIONS

Pub. No.:    WO/2014/197986    International Application No.:    PCT/CA2014/050539
Publication Date: Fri Dec 19 00:59:59 CET 2014 International Filing Date: Wed Jun 11 01:59:59 CEST 2014
IPC: C12Q 1/68
C07K 14/47
C12Q 1/02
G01N 33/567
G01N 33/68
C07K 14/71
Applicants: THE GOVERNING COUNCIL OF THE UNIVERSITY OF TORONTO
Inventors: STAGLJAR, Igor
PETSCHNIGG, Julia
GROISMAN, Bella
Title: DETECTION OF PROTEIN TO PROTEIN INTERACTIONS
Abstract:
The present invention relates to methods and kits for detecting the interaction between a first membrane protein and a second protein in mammalian cells as well as to identify molecules that can disrupt protein to protein interactions. The invention relies on the functional reconstitution of an active human ubiquitin by two inactive fragments upon the interaction of two proteins attached through a linker to the inactive fragments. The reconstituted ubiquitin is then cleaved by human ubiquitin proteases resulting in the release of an artificial transcription factor, which in turn activates a reporter gene transcription. Activation of the reporter gene is indicative of the interaction between the two proteins.