Universitéde Montreal

The Royal Institution for the Advancement of Learning / Mcgill University

Domain Therapeutics

Mireille Hogue

Michel Bouvier

Viktoriya Lukasheva

Christian Le Gouill

Stephane Alain Laporte

Billy Breton

Hiroyuki Kobayashi

Yoon Namkung

Denis Deblois

Stéphane Alain Laporte

Étienne Durette

Etienne Durette

Resonance energy transfer (RET)- or protein-fragment complement assay (PCA)-based biosensors useful for assessing the activity of G-proteins are described. These biosensors are based on the competition between the G&#x3b1; subunit and a G&#x3b2;&#x3b3; interacting protein (&#x3b2;&#x3b3; IP) for the binding to the G&#x3b2;&#x3b3; dimer. These biosensors comprises (1) a &#x3b2;&#x3b3; IP and (2) a G&#x3b2; or G&#x3b3; protein; a GPCR; or a plasma membrane targeting domain, fused to suitable RET or PCA tags. Methods using such biosensors for different applications, including the identification of agents that modulates G-protein activity or for the characterization of GPCR signaling/regulation, such as G-protein preferences and activation profiles of GPCRs, are also described.

Biosensor based on G&#x3b2;&#x3b3;-interacting proteins to monitor G-protein activation

Bioluminescence resonance energy transfer (BRET) biosensors for assessing the intracellular localization, internalization and trafficking into cellular compartments of proteins such as receptors, and other biomolecules such as second messengers, are disclosed. These biosensors, which are dependent on the concentration/density of the BRET donor and acceptor in cellular compartments rather that specific protein-protein interactions, use aGFP/Luc BRET pair, which allows the robust and reproducible monitoring of protein trafficking/localization, with a sensitivity compatible with high-throughput screening (HTS). The use of these biosensors for various applications, including assessing/monitoring protein endocytosis, recycling and intracellular trafficking, receptor maturation/rescue by pharmacological chaperones, various endocytosis/exocytosis processes, activation/inhibition, as well as biomolecule concentration/density in different cellular compartments, is also disclosed.

Renilla based biosensors for monitoring biomolecule localization and trafficking in cells

Growing community of inventors

Laval, Canada

Mireille Hogue