In vivo quantification of protein-protein interactions in Saccharomyces cerevisiae using bimolecular fluorescence complementation assay

J Microbiol Methods. 2010 Nov;83(2):194-201. doi: 10.1016/j.mimet.2010.08.021. Epub 2010 Sep 7.

Abstract

Most of the biological processes are carried out and regulated by dynamic networks of protein-protein interactions. In this study, we demonstrate the feasibility of the bimolecular fluorescence complementation (BiFC) assay for in vivo quantitative analysis of protein-protein interactions in Saccharomyces cerevisiae. We show that the BiFC assay can be used to quantify not only the amount but also the cell-to-cell variation of protein-protein interactions in S. cerevisiae. In addition, we show that protein sumoylation and condition-specific protein-protein interactions can be quantitatively analyzed by using the BiFC assay. Taken together, our results validate that the BiFC assay is a very effective method for quantitative analysis of protein-protein interactions in living yeast cells and has a great potential as a versatile tool for the study of protein function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Fluorescence
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Protein Binding
  • Protein Interaction Mapping / methods*
  • Proteomics / methods*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Bacterial Proteins
  • Luminescent Proteins
  • Saccharomyces cerevisiae Proteins
  • yellow fluorescent protein, Bacteria