The mapping of yeast's G-protein coupled receptor with an atomic force microscope

Nanoscale. 2015 Mar 21;7(11):4956-63. doi: 10.1039/c4nr05940a.

Abstract

An atomic force microscope (AFM) can measure the adhesion force between a sample and a cantilever while simultaneously applying a rupture force during the imaging of a sample. An AFM should be useful in targeting specific proteins on a cell surface. The present study proposes the use of an AFM to measure the adhesion force between targeting receptors and their ligands, and to map the targeting receptors. In this study, Ste2p, one of the G protein-coupled receptors (GPCRs), was chosen as the target receptor. The specific force between Ste2p on a yeast cell surface and a cantilever modified with its ligand, α-factor, was measured and found to be approximately 250 pN. In addition, through continuous measuring of the cell surface, a mapping of the receptors on the cell surface could be performed, which indicated the differences in the Ste2p expression levels. Therefore, the proposed AFM system is accurate for cell diagnosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbocyanines / chemistry
  • Ligands
  • Mating Factor
  • Microscopy, Atomic Force*
  • Peptides / chemistry
  • Peptides / genetics
  • Peptides / metabolism
  • Receptors, Mating Factor / antagonists & inhibitors
  • Receptors, Mating Factor / genetics
  • Receptors, Mating Factor / metabolism*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / antagonists & inhibitors
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Carbocyanines
  • Ligands
  • Peptides
  • Receptors, Mating Factor
  • STE2 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Mating Factor