title: Direct identification of ligand-receptor interactions on living cells and tissues.
abstract: 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. 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.
authors.affiliation: Department of Biology, Institute of Molecular Systems Biology, ETH Zurich, Switzerland.
authors.forename: Andreas P
authors.initials: AP
authors.lastname: Frei
authors.forename: Ock-Youm
authors.initials: OY
authors.lastname: Jeon
authors.forename: Samuel
authors.initials: S
authors.lastname: Kilcher
authors.forename: Hansjoerg
authors.initials: H
authors.lastname: Moest
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Henning
authors.forename: Christian
authors.initials: C
authors.lastname: Jost
authors.forename: Andreas
authors.initials: A
authors.lastname: Plückthun
authors.forename: Jason
authors.initials: J
authors.lastname: Mercer
authors.forename: Ruedi
authors.initials: R
authors.lastname: Aebersold
authors.forename: Erick M
authors.initials: EM
authors.lastname: Carreira
authors.forename: Bernd
authors.initials: B
authors.lastname: Wollscheid
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D006834:Hydrazines / Q000737:chemistry; D007328:Insulin / Q000494:pharmacology; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000187:drug effects; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D040901:Proteomics; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013388:Succinimides / Q000737:chemistry
chemical_list: D006834:Hydrazines; D007328:Insulin; D008024:Ligands; D010455:Peptides; D011956:Receptors, Cell Surface; D013388:Succinimides; C585574:TRICEPS compound; D001710:Biotin