RTK LinkLight™ assay
BioInvenu’s proprietary LinkLight™ technology offers innovative tools for cell based protein-protein interaction assays.
Receptor tyrosine kinases (RTK) are single transmembrane cell surface proteins. The extracellular N-terminal region can bind to extracellular ligands, such as growth factors or hormones. The intracellular C-terminal region comprises the catalytic domain which is responsible for receptor autophosphorylation and tyrosine phosphorylation of downstream substrates. RTKs not only are key regulators of normal cellular processes but also play a critical role in the development and progression of human cancer.
The assay is based on activated RTKs interacting with a protein containing phosphotyrosine binding (PTB) domain. An inactive permuted luciferase (pLuc) containing a Tobacco Etch Virus (TEV) protease cleavage site is fused to a PTB domain (PTB-pLuc) and a full-length RTK is fused to the TEV protease (RTK-TEV). Active luciferase is generated when the PTB -pLuc fusion protein interacts with the full-length RTK fused to TEV protease upon ligand activation.
RTK LinkLight™ assay offers a novel cell-based screening method using full-length RTK proteins in its native confirmation. The assay could identify compounds that block RTK- ligand interaction, RTK-PTB interaction, kinase activity, or modulate RTK conformation change resulting in signal alternation. In addition, RTK LinkLight™ assay could be used as a screen method to find biologics block RTK function.