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Large-scale analysis of diffusional dynamics of proteins in living yeast cells using fluorescence correlation spectroscopy.

Authors
  • Fukuda, Takafumi1
  • Kawai-Noma, Shigeko2
  • Pack, Chan-Gi3
  • Taguchi, Hideki4
  • 1 School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan. , (Japan)
  • 2 Department of Applied Chemistry and Biotechnology, Chiba University, Chiba, Japan. , (Japan)
  • 3 Asan Institute for Life Sciences, Asan Medical Center, Seoul, 05505, South Korea. , (North Korea)
  • 4 School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan; Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama, Japan. Electronic address: [email protected] , (Japan)
Type
Published Article
Journal
Biochemical and Biophysical Research Communications
Publisher
Elsevier
Publication Date
Dec 03, 2019
Volume
520
Issue
2
Pages
237–242
Identifiers
DOI: 10.1016/j.bbrc.2019.09.066
PMID: 31594638
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

In the living cells, the majority of proteins does not work alone, but interact with other proteins or other biomolecules to maintain the cellular function, constituting a "protein community". Previous efforts on mass spectroscopy-based protein interaction networks, interactomes, have provided a picture on the protein community. However, these were static information after cells were disrupted. For a better understanding of the protein community in cells, it is important to know the properties of intracellular dynamics and interactions. Since hydrodynamic size and mobility of proteins are related into such properties, direct measurement of diffusional motion of proteins in single living cells will be helpful for uncovering the properties. Here we completed measurement of the diffusion and homo-oligomeric properties of 369 cytoplasmic GFP-fusion proteins in living yeast Saccharomyces cerevisiae cells using fluorescence correlation spectroscopy (FCS). The large-scale analysis showed that the motions of majority of proteins obeyed a two-component (i.e. slow and fast components) diffusion model. Remarkably, both of the two components diffused more slowly than expected monomeric states. In addition, further analysis suggested that more proteins existed as homo-oligomeric states in living cells than previously expected. Our study, which characterizes the dynamics of proteins in living cells on a large-scale, provided a global view on intracellular protein dynamics to understand the protein community. Copyright © 2019 Elsevier Inc. All rights reserved.

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