Biophysical Analysis of Lipid Domains in Mammalian and Yeast Membranes by Fluorescence Spectroscopy
| Autor: | Andreia Bento-Oliveira, Beatriz Mestre, Rodrigo F.M. de Almeida, Carla A. Sousa, Liana C. Silva, Filipa C. Santos |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
030103 biophysics Liposome biology Chemistry Saccharomyces cerevisiae biology.organism_classification Fluorescence spectroscopy Yeast 03 medical and health sciences 030104 developmental biology Membrane Förster resonance energy transfer Biophysics Fluorescence microscope lipids (amino acids peptides and proteins) Sphingomyelin |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071608135 |
| DOI: | 10.1007/978-1-0716-0814-2_14 |
| Popis: | The use of steady-state and time-resolved fluorescence spectroscopy to study sterol and sphingolipid-enriched lipid domains as diverse as the ones found in mammalian and fungal membranes is herein described. We first address how to prepare liposomes that mimic raft-containing membranes of mammalian cells and how to use fluorescence spectroscopy to characterize the biophysical properties of these membrane model systems. We further illustrate the application of Forster resonance energy transfer (FRET) to study nanodomain reorganization upon interaction with small bioactive molecules, phenolic acids, an important group of phytochemical compounds. This methodology overcomes the resolution limits of conventional fluorescence microscopy allowing for the identification and characterization of lipid domains at the nanoscale.We continue by showing how to use fluorescence spectroscopy in the biophysical analysis of more complex biological systems, namely the plasma membrane of Saccharomyces cerevisiae yeast cells and the necessary adaptations to the filamentous fungus Neurospora crassa , evaluating the global order of the membrane, sphingolipid-enriched domains rigidity and abundance, and ergosterol-dependent properties. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|