Abstract
Supported lipid bilayers are a well-developed model system for the study of membranes and their associated proteins, such as membrane channels, enzymes, and receptors. These versatile model membranes can be made from various components, ranging from simple synthetic phospholipids to complex mixtures of constituents, mimicking the cell membrane with its relevant physiochemical and molecular phenomena. In addition, the high stability of supported lipid bilayers allows for their study via a wide array of experimental probes. In this work, we describe a platform for supported lipid bilayers that is accessible both electrically and optically, and demonstrate direct optical observation of the transmembrane potential of supported lipid bilayers. We show that the polarization of the supported membrane can be electrically controlled and optically probed using voltage-sensitive dyes. Membrane polarization dynamics is understood through electrochemical impedance spectroscopy and the analysis of an equivalent electrical circuit model. In addition, we describe the effect of the conducting electrode layer on the fluorescence of the optical probe through metal-induced energy transfer, and show that while this energy transfer has an adverse effect on the voltage sensitivity of the fluorescent probe, its strong distance dependency allows for axial localization of fluorescent emitters with ultrahigh accuracy. We conclude with a discussion on possible applications of this platform for the study of voltage-dependent membrane proteins and other processes in membrane biology and surface science.
Original language | English |
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Pages (from-to) | 2624-2637 |
Number of pages | 14 |
Journal | Biophysical Journal |
Volume | 121 |
Issue number | 13 |
DOIs | |
State | Published - 5 Jul 2022 |
Bibliographical note
Publisher Copyright:© 2022 Biophysical Society
Funding
We thank Dr. Shay Tirosh and Prof. Arie Zaban for potentiostat instrument access and help with EIS, Dr. Maria Tkachev and Dr. Ilana Perelshtein for performing cross-sectional HR-TEM imaging of the electrode, Dr. Ayelet Atkins for performing cryo-EM imaging of the vesicles, and Mark Oksman for performing ellipsometry measurements. This work has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program under grant agreement no. 669941, by the Human Frontier Science Program (HFSP) research grant RGP0061/2015, by the BER program of the Department of Energy Office of Science grant DE-SC0020338, by the STROBE National Science Foundation Science & Technology Center, grant no. DMR-1548924, by the Israel Science Foundation (ISF) grant 813/19, and by the Bar-Ilan Research & Development Co, the Israel Innovation Authority, grant no. 63392. The authors declare no competing interests. We thank Dr. Shay Tirosh and Prof. Arie Zaban for potentiostat instrument access and help with EIS, Dr. Maria Tkachev and Dr. Ilana Perelshtein for performing cross-sectional HR-TEM imaging of the electrode, Dr. Ayelet Atkins for performing cryo-EM imaging of the vesicles, and Mark Oksman for performing ellipsometry measurements. This work has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program under grant agreement no. 669941 , by the Human Frontier Science Program (HFSP) research grant RGP0061/2015 , by the BER program of the Department of Energy Office of Science grant DE-SC0020338 , by the STROBE National Science Foundation Science & Technology Center, grant no. DMR-1548924 , by the Israel Science Foundation (ISF) grant 813/19 , and by the Bar-Ilan Research & Development Co , the Israel Innovation Authority , grant no. 63392 .
Funders | Funder number |
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Israel Innovation Authority | 63392 |
STROBE National Science Foundation Science & Technology Center | DMR-1548924 |
Office of Science | DE-SC0020338 |
Horizon 2020 Framework Programme | |
European Commission | |
Human Frontier Science Program | RGP0061/2015 |
Israel Science Foundation | 813/19 |
Horizon 2020 | 669941 |