Correlating single-molecule characteristics of the yeast aquaglyceroporin Fps1 with environmental perturbations directly in living cells
Artikel i vetenskaplig tidskrift, 2020

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.

Single-molecule

Plasma membrane channels

Osmotic condition

Super-resolution

Cell stress

Living cell

Författare

Sviatlana Shashkova

University of York

Mikael Andersson

Göteborgs universitet

Stefan Hohmann

Chalmers, Biologi och bioteknik

Mark C. Leake

University of York

Methods

1046-2023 (ISSN) 1095-9130 (eISSN)

Vol. In Press

Ämneskategorier

Cellbiologi

Biofysik

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

DOI

10.1016/j.ymeth.2020.05.003

PubMed

32387484

Mer information

Senast uppdaterat

2020-12-29