Cargando…
Intracellular Protein–Lipid Interactions Studied by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy
[Image: see text] Protein–membrane interactions play key roles in essential cellular processes; studying these interactions in the cell is a challenging task of modern biophysical chemistry. A prominent example is the interaction of human α-synuclein (αS) with negatively charged membranes. It has be...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2021
|
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7957861/ https://www.ncbi.nlm.nih.gov/pubmed/33663214 http://dx.doi.org/10.1021/acs.jpclett.0c03583 |
_version_ | 1783664744395177984 |
---|---|
author | Braun, Theresa S. Stehle, Juliane Kacprzak, Sylwia Carl, Patrick Höfer, Peter Subramaniam, Vinod Drescher, Malte |
author_facet | Braun, Theresa S. Stehle, Juliane Kacprzak, Sylwia Carl, Patrick Höfer, Peter Subramaniam, Vinod Drescher, Malte |
author_sort | Braun, Theresa S. |
collection | PubMed |
description | [Image: see text] Protein–membrane interactions play key roles in essential cellular processes; studying these interactions in the cell is a challenging task of modern biophysical chemistry. A prominent example is the interaction of human α-synuclein (αS) with negatively charged membranes. It has been well-studied in vitro, but in spite of the huge amount of lipid membranes in the crowded environment of biological cells, to date, no interactions have been detected in cells. Here, we use rapid-scan (RS) electron paramagnetic resonance (EPR) spectroscopy to study αS interactions with negatively charged vesicles in vitro and upon transfection of the protein and lipid vesicles into model cells, i.e., oocytes of Xenopus laevis. We show that protein–vesicle interactions are reflected in RS spectra in vitro and in cells, which enables time-resolved monitoring of protein–membrane interaction upon transfection into cells. Our data suggest binding of a small fraction of αS to endogenous membranes. |
format | Online Article Text |
id | pubmed-7957861 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-79578612021-03-16 Intracellular Protein–Lipid Interactions Studied by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy Braun, Theresa S. Stehle, Juliane Kacprzak, Sylwia Carl, Patrick Höfer, Peter Subramaniam, Vinod Drescher, Malte J Phys Chem Lett [Image: see text] Protein–membrane interactions play key roles in essential cellular processes; studying these interactions in the cell is a challenging task of modern biophysical chemistry. A prominent example is the interaction of human α-synuclein (αS) with negatively charged membranes. It has been well-studied in vitro, but in spite of the huge amount of lipid membranes in the crowded environment of biological cells, to date, no interactions have been detected in cells. Here, we use rapid-scan (RS) electron paramagnetic resonance (EPR) spectroscopy to study αS interactions with negatively charged vesicles in vitro and upon transfection of the protein and lipid vesicles into model cells, i.e., oocytes of Xenopus laevis. We show that protein–vesicle interactions are reflected in RS spectra in vitro and in cells, which enables time-resolved monitoring of protein–membrane interaction upon transfection into cells. Our data suggest binding of a small fraction of αS to endogenous membranes. American Chemical Society 2021-03-05 2021-03-11 /pmc/articles/PMC7957861/ /pubmed/33663214 http://dx.doi.org/10.1021/acs.jpclett.0c03583 Text en © 2021 The Authors. Published by American Chemical Society This is an open access article published under an ACS AuthorChoice License (https://creativecommons.org/licenses/by/4.0/) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes. |
spellingShingle | Braun, Theresa S. Stehle, Juliane Kacprzak, Sylwia Carl, Patrick Höfer, Peter Subramaniam, Vinod Drescher, Malte Intracellular Protein–Lipid Interactions Studied by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy |
title | Intracellular Protein–Lipid Interactions Studied
by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy |
title_full | Intracellular Protein–Lipid Interactions Studied
by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy |
title_fullStr | Intracellular Protein–Lipid Interactions Studied
by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy |
title_full_unstemmed | Intracellular Protein–Lipid Interactions Studied
by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy |
title_short | Intracellular Protein–Lipid Interactions Studied
by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy |
title_sort | intracellular protein–lipid interactions studied
by rapid-scan electron paramagnetic resonance spectroscopy |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7957861/ https://www.ncbi.nlm.nih.gov/pubmed/33663214 http://dx.doi.org/10.1021/acs.jpclett.0c03583 |
work_keys_str_mv | AT brauntheresas intracellularproteinlipidinteractionsstudiedbyrapidscanelectronparamagneticresonancespectroscopy AT stehlejuliane intracellularproteinlipidinteractionsstudiedbyrapidscanelectronparamagneticresonancespectroscopy AT kacprzaksylwia intracellularproteinlipidinteractionsstudiedbyrapidscanelectronparamagneticresonancespectroscopy AT carlpatrick intracellularproteinlipidinteractionsstudiedbyrapidscanelectronparamagneticresonancespectroscopy AT hoferpeter intracellularproteinlipidinteractionsstudiedbyrapidscanelectronparamagneticresonancespectroscopy AT subramaniamvinod intracellularproteinlipidinteractionsstudiedbyrapidscanelectronparamagneticresonancespectroscopy AT dreschermalte intracellularproteinlipidinteractionsstudiedbyrapidscanelectronparamagneticresonancespectroscopy |