Direct identification of ligand-receptor interactions on living cells and tissues

September 16, 2012

Reference

Nature Biotechnology 30, 997–1001 (2012) doi:10.1038/nbt.2354 – Received06 April 2012 Accepted08 August 2012 Published online16 September 2012 Many cellular responses are triggered by proteins, drugs or pathogens binding to cell-surface receptors, but it can be challenging to identify which receptors are bound by a given ligand. Here we describe TRICEPS, a chemoproteomic reagent with three moieties—one that binds ligands containing an amino group, a second that binds glycosylated receptors on living cells and a biotin tag for purifying the receptor peptides for identification by quantitative mass spectrometry.

Abstract

We validated this ligand-based, receptor-capture (LRC) technology using insulin, transferrin, apelin, epidermal growth factor, the therapeutic antibody trastuzumab and two DARPins targeting ErbB2. In some cases, we could also determine the approximate ligand-binding sites on the receptors. Using TRICEPS to label intact mature vaccinia viruses, we identified the cell surface proteins AXL, M6PR, DAG1, CSPG4 and CDH13 as binding factors on human cells. This technology enables the identification of receptors for many types of ligands under near-physiological conditions and without the need for genetic manipulations.

Author

Andreas P Frei, Ock-Youm Jeon, Samuel Kilcher, Hansjoerg Moest, Lisa M Henning, Christian Jost, Andreas Plückthun, Jason Mercer, Ruedi Aebersold, Erick M Carreira & Bernd Wollscheid