title: Cell wall trapping of autocrine peptides for human G-protein-coupled receptors on the yeast cell surface.
abstract: G-protein-coupled receptors (GPCRs) regulate a wide variety of physiological processes and are important pharmaceutical targets for drug discovery. Here, we describe a unique concept based on yeast cell-surface display technology to selectively track eligible peptides with agonistic activity for human GPCRs (Cell Wall Trapping of Autocrine Peptides (CWTrAP) strategy). In our strategy, individual recombinant yeast cells are able to report autocrine-positive activity for human GPCRs by expressing a candidate peptide fused to an anchoring motif. Following expression and activation, yeast cells trap autocrine peptides onto their cell walls. Because captured peptides are incapable of diffusion, they have no impact on surrounding yeast cells that express the target human GPCR and non-signaling peptides. Therefore, individual yeast cells can assemble the autonomous signaling complex and allow single-cell screening of a yeast population. Our strategy may be applied to identify eligible peptides with agonistic activity for target human GPCRs.
authors.affiliation: Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ishii
authors.forename: Nobuo
authors.initials: N
authors.lastname: Yoshimoto
authors.forename: Kenji
authors.initials: K
authors.lastname: Tatematsu
authors.forename: Shun'ichi
authors.initials: S
authors.lastname: Kuroda
authors.forename: Chiaki
authors.initials: C
authors.lastname: Ogino
authors.forename: Hideki
authors.initials: H
authors.lastname: Fukuda
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D019898:Autocrine Communication / Q000502:physiology*; D001681:Biological Assay / Q000379:methods*; D001709:Biotechnology / Q000379:methods; D015153:Blotting, Western; D002473:Cell Wall / Q000378:metabolism*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D010455:Peptides / Q000302:isolation & purification* / Q000378:metabolism*; D049049:Pheromones, Human / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000378:metabolism*; D012441:Saccharomyces cerevisiae
chemical_list: D010455:Peptides; D049049:Pheromones, Human; D043562:Receptors, G-Protein-Coupled