Tracking Global and Local Changes in Membrane Fluidity Through Fluorescence Spectroscopy and Microscopy

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2022 Nov 29

PMID 36445586

Authors

Madeleine Humphrey

Ireny Abdelmesseh Nekhala

Kathi Scheinpflug

Oxana Krylova

Ann-Britt Schafer

Jessica A Buttress

Michaela Wenzel

Henrik Strahl

Affiliations

Soon will be listed here.

Abstract

Membrane fluidity is a critical parameter of cellular membranes, which cells continuously strive to maintain within a viable range. Interference with the correct membrane fluidity state can strongly inhibit cell function. Triggered changes in membrane fluidity and associated impacts on lipid domains have been postulated to contribute to the mechanism of action of membrane targeting antimicrobials, but the corresponding analyses have been hampered by the absence of readily available analytical tools. Here, we expand upon the protocols outlined in the first edition of this book, providing further and alternative protocols that can be used to measure changes in membrane fluidity. We provide detailed protocols, which allow straightforward in vivo and in vitro measurement of antibiotic compound-triggered changes in membrane fluidity and fluid membrane microdomains. Furthermore, we summarize useful strains constructed by us and others to characterize and confirm lipid specificity of membrane antimicrobials directly in vivo.

Citing Articles

Lipid phase separation impairs membrane thickness sensing by the sensor kinase DesK.

Sidarta M, Lorente Martin A, Monsalve A, Marinho Righetto G, Schafer A, Wenzel M Microbiol Spectr. 2024; 12(6):e0392523.

PMID: 38717171 PMC: 11237406. DOI: 10.1128/spectrum.03925-23.

Dissecting antibiotic effects on the cell envelope using bacterial cytological profiling: a phenotypic analysis starter kit.

Schafer A, Sidarta M, Abdelmesseh Nekhala I, Marinho Righetto G, Arshad A, Wenzel M Microbiol Spectr. 2024; :e0327523.

PMID: 38289933 PMC: 10913488. DOI: 10.1128/spectrum.03275-23.

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