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Correlating single-molecule characteristics of the yeast aquaglyceroporin Fps1 with environmental perturbations directly in living cells.

Authors
  • Shashkova, Sviatlana1
  • Andersson, Mikael2
  • Hohmann, Stefan3
  • Leake, Mark C4
  • 1 Department of Physics, University of York, YO10 5DD York, UK. Electronic address: [email protected]
  • 2 Department of Chemistry and Molecular Biology, University of Gothenburg, 405 30 Gothenburg, Sweden. Electronic address: [email protected] , (Sweden)
  • 3 Department of Biology and Biological Engineering, Chalmers University of Technology, 412 96 Gothenburg, Sweden. Electronic address: [email protected] , (Sweden)
  • 4 Department of Physics, University of York, YO10 5DD York, UK. Electronic address: [email protected]
Type
Published Article
Journal
Methods
Publisher
Elsevier
Publication Date
Sep 01, 2021
Volume
193
Pages
46–53
Identifiers
DOI: 10.1016/j.ymeth.2020.05.003
PMID: 32387484
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Membrane proteins play key roles at the interface between the cell and its environment by mediating selective import and export of molecules via plasma membrane channels. Despite a multitude of studies on transmembrane channels, understanding of their dynamics directly within living systems is limited. To address this, we correlated molecular scale information from living cells with real time changes to their microenvironment. We employed super-resolved millisecond fluorescence microscopy with a single-molecule sensitivity, to track labelled molecules of interest in real time. We use as example the aquaglyceroporin Fps1 in the yeast Saccharomyces cerevisiae to dissect and correlate its stoichiometry and molecular turnover kinetics with various extracellular conditions. We show that Fps1 resides in multi tetrameric clusters while hyperosmotic and oxidative stress conditions cause Fps1 reorganization. Moreover, we demonstrate that rapid exposure to hydrogen peroxide causes Fps1 degradation. In this way we shed new light on aspects of architecture and dynamics of glycerol-permeable plasma membrane channels. Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.

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