title: A proximity based general method for identification of ligand and receptor interactions in living cells.
abstract: Autocrine based selections from intracellular combinatorial antibody and peptide libraries have proven to be a powerful method for selection of agonists and identification of new therapeutic targets. However, success requires a case-by-case construction of a robust selection system which is a process that can be time consuming and expensive. Here we report a general system that takes advantage of the chemical rate acceleration caused by approximation of a membrane tethered ligand and its receptor. The system uses an artificial signal transduction and is, thus, agnostic to the endogenous signal transduction of the receptor-ligand system. This method allows analysis of receptor-ligand interactions and selection of molecules from large libraries that interact with receptors when they are in their natural milieu.
authors.affiliation: Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, United States. Electronic address: hongkai@scripps.edu.
authors.forename: Hongkai
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, United States.
authors.forename: Jia
authors.initials: J
authors.lastname: Xie
authors.affiliation: Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, United States. Electronic address: rlerner@scripps.edu.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Lerner
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D020650:Combinatorial Chemistry Techniques; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D000067757:Glucagon-Like Peptide-1 Receptor; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D019151:Peptide Library; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*; D018027:Receptors, Glucagon / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: C000593833:GLP1R protein, human; D000067757:Glucagon-Like Peptide-1 Receptor; D008024:Ligands; D019151:Peptide Library; D011956:Receptors, Cell Surface; D018027:Receptors, Glucagon; D011993:Recombinant Fusion Proteins; D010450:Endopeptidases; C099488:TEV protease
keywords: Effective molarity; Proximity based method; Receptor–ligand interaction; Universal reporter system