Intracellular Protein-Lipid Interactions Studied by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy

Theresa S. Braun; Juliane Stehle; Sylwia Kacprzak; Patrick Carl; Peter Höfer; Vinod Subramaniam; Malte Drescher

doi:10.1021/acs.jpclett.0c03583

Intracellular Protein-Lipid Interactions Studied by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy

Theresa S. Braun, Juliane Stehle, Sylwia Kacprzak, Patrick Carl, Peter Höfer, Vinod Subramaniam, Malte Drescher^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

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.

Original language	English
Pages (from-to)	2471-2475
Number of pages	5
Journal	Journal of Physical Chemistry Letters
Volume	12
Issue number	9
Early online date	5 Mar 2021
DOIs	https://doi.org/10.1021/acs.jpclett.0c03583
Publication status	Published - 11 Mar 2021

Bibliographical note

Funding Information:
We gratefully acknowledge the experimental contributions of Martina Adam-Wels, including protein expression and purification. Plasmid pET11C_ASYN_Δ2–11 encoding αS Δ2–11 was kindly provided by Christiaan Karreman from the group of Marcel Leist, University of Konstanz. This work was supported financially by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (Grant Agreement No. 772027-SPICEERC-2017-COG) and from the Deutsche Forschungsgemeinschaft (SFB 969, Project C03).

Publisher Copyright:
©

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Funding

We gratefully acknowledge the experimental contributions of Martina Adam-Wels, including protein expression and purification. Plasmid pET11C_ASYN_Δ2–11 encoding αS Δ2–11 was kindly provided by Christiaan Karreman from the group of Marcel Leist, University of Konstanz. This work was supported financially by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (Grant Agreement No. 772027-SPICEERC-2017-COG) and from the Deutsche Forschungsgemeinschaft (SFB 969, Project C03).

Funders	Funder number
Horizon 2020 Framework Programme	772027
European Research Council
Deutsche Forschungsgemeinschaft	SFB 969

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title = "Intracellular Protein-Lipid Interactions Studied by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy",

abstract = "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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N2 - 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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Intracellular Protein-Lipid Interactions Studied by Rapid-Scan Electron Paramagnetic Resonance Spectroscopy

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