Improved identification of agonist-mediated Gα(i)-specific human G-protein-coupled receptor signaling in yeast cells by flow cytometry

Anal Biochem. 2012 Jul 15;426(2):129-33. doi: 10.1016/j.ab.2012.04.012. Epub 2012 Apr 11.

Abstract

Flow cytometry enables comparative quantification, population analysis, and high-throughput screening of agonist-mediated G-protein-coupled receptor (GPCR) signaling in genetically engineered yeasts. By using flow cytometry, we found that transformation of yeast cells with a low plasmid number is critical both for the construction of large screening libraries and for stable signal transmission in cell ensembles. Based on these findings, we constructed an engineered yeast strain for the improved identification of signal promotion by Gα(i)-specific human GPCRs using flow cytometry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Enzyme Inhibitors / analysis*
  • Flow Cytometry*
  • GTP-Binding Protein alpha Subunits, Gi-Go / agonists*
  • GTP-Binding Protein alpha Subunits, Gi-Go / metabolism
  • Humans
  • Plasmids / metabolism
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Somatostatin / genetics
  • Receptors, Somatostatin / metabolism
  • Saccharomyces cerevisiae / metabolism*
  • Signal Transduction

Substances

  • Enzyme Inhibitors
  • Receptors, G-Protein-Coupled
  • Receptors, Somatostatin
  • somatostatin receptor 5
  • GTP-Binding Protein alpha Subunits, Gi-Go