Blog
About

18
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Cell Wall Trapping of Autocrine Peptides for Human G-Protein-Coupled Receptors on the Yeast Cell Surface

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          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.

          Related collections

          Most cited references 41

          • Record: found
          • Abstract: found
          • Article: not found

          Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications.

          A set of yeast strains based on Saccharomyces cerevisiae S288C in which commonly used selectable marker genes are deleted by design based on the yeast genome sequence has been constructed and analysed. These strains minimize or eliminate the homology to the corresponding marker genes in commonly used vectors without significantly affecting adjacent gene expression. Because the homology between commonly used auxotrophic marker gene segments and genomic sequences has been largely or completely abolished, these strains will also reduce plasmid integration events which can interfere with a wide variety of molecular genetic applications. We also report the construction of new members of the pRS400 series of vectors, containing the kanMX, ADE2 and MET15 genes.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis.

            The functions of many open reading frames (ORFs) identified in genome-sequencing projects are unknown. New, whole-genome approaches are required to systematically determine their function. A total of 6925 Saccharomyces cerevisiae strains were constructed, by a high-throughput strategy, each with a precise deletion of one of 2026 ORFs (more than one-third of the ORFs in the genome). Of the deleted ORFs, 17 percent were essential for viability in rich medium. The phenotypes of more than 500 deletion strains were assayed in parallel. Of the deletion strains, 40 percent showed quantitative growth defects in either rich or minimal medium.
              Bookmark
              • Record: found
              • Abstract: not found
              • Article: not found

              Improved method for high efficiency transformation of intact yeast cells.

               A Jean,  John Woods,  R Gietz (1992)
                Bookmark

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                18 May 2012
                : 7
                : 5
                Affiliations
                [1 ]Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, Japan
                [2 ]Department of Structural Molecular Biology, Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka, Japan
                [3 ]Graduate School of Bioagricultural Sciences, Nagoya University, Furo, Chikusa, Nagoya, Japan
                [4 ]Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, Japan
                University of Connecticut, United States of America
                Author notes

                Conceived and designed the experiments: JI NY KT SK CO HF AK. Performed the experiments: JI. Analyzed the data: JI. Contributed reagents/materials/analysis tools: JI. Wrote the paper: JI AK.

                PONE-D-11-25420
                10.1371/journal.pone.0037136
                3356411
                22623985
                Ishii et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                Counts
                Pages: 10
                Categories
                Research Article
                Biology
                Biochemistry
                Cytochemistry
                Cell Membrane
                Membrane Proteins
                Proteins
                Transmembrane Proteins
                Biomacromolecule-Ligand Interactions
                Drug Discovery
                Biophysics
                Biomacromolecule-Ligand Interactions
                Biotechnology
                Bioengineering
                Biological Systems Engineering
                Biomimetics
                Genetic Engineering
                Genetically Modified Organisms
                Applied Microbiology
                Drug Discovery
                Chemistry
                Analytical Chemistry
                Chemical Analysis
                Colorimetric Analysis
                Chemical Biology
                Engineering
                Bioengineering
                Biological Systems Engineering
                Biomimetics

                Uncategorized

                Comments

                Comment on this article