Ligand nanobeads to activate, identify and assay the insulin receptor complex on cultured macrophages

The protein hormones (ligands) that circulate in blood plasma and their receptors on the surface of cells are among the most important diagnostic and therapeutic molecules and are a key focus for the detection and treatment of diseases. Presenting the ligands on chromatography beads may permit the isolation of receptors by a modified form of affinity chromatography. The binding of the hormone insulin activates the insulin receptor on the cell surface which in turn leads to the recruitment of accessory enzymes or signalling proteins to the activated receptor complex that may be new drug targets. Finding new receptor specific drug targets is crucially important information that to date has eluded genetic, DNA/RNA or biotin-based technologies and may lead to new treatments for disease and is of great importance. Affinity chromatography won the Wolf prize for Pedro Cuatrecasas and Meir Wilchek in 1987. Electrospray won the Nobel Prize for John Fenn in 2002. Ion trap mass spectrometry won the Nobel prize for Wolfgang Paul in 1989. The combination of affinity chromatography, electrospray ionization and ion trap mass now permit the direct analysis of cell surface receptor complexes to reveal new drug targets. Here we propose a general solution for discovering the activated receptor complex from the cell surface to reveal the drug target proteins complexed with the receptor and test the effect of new molecular medicines termed silencing RNA, as well as traditional drugs, on the function of the receptors from live cells under the laser scanning confocal microscope.

Faculty Supervisor:

John Marshall

Student:

Partner:

YYZ Pharmatech Inc.

Discipline:

Life Sciences

Sector:

Health and Related Sciences & Technology; Professional, scientific and technical services

University:

Toronto Metropolitan University

Program: