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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...

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Autores principales: Braun, Theresa S., Stehle, Juliane, Kacprzak, Sylwia, Carl, Patrick, Höfer, Peter, Subramaniam, Vinod, Drescher, Malte
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
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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.
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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
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