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Issue 20, 2014
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Voltage-dependent structural changes of the membrane-bound anion channel hVDAC1 probed by SEIRA and electrochemical impedance spectroscopy

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Abstract

The voltage-dependent anion channel (VDAC) is a transmembrane protein that regulates the transfer of metabolites between the cytosol and the mitochondrium. Opening and partial closing of the channel is known to be driven by the transmembrane potential via a mechanism that is not fully understood. In this work, we employed a spectroelectrochemical approach to probe the voltage-induced molecular structure changes of human VDAC1 (hVDAC1) embedded in a tethered bilayer lipid membrane on a nanostructured Au electrode. The model membrane consisted of a mixed self-assembled monolayer of 6-mercaptohexanol and (cholesterylpolyethylenoxy)thiol, followed by the deposition of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine vesicles including hVDAC1. The stepwise assembly of the model membrane and the incorporation of hVDAC1 were monitored by surface enhanced infrared absorption and electrochemical impedance spectroscopy. Difference spectra allowed for identifying the spectral changes which may be associated with the transition from the open to the “closed” states by shifting the potential above or below the transmembrane potential determined to be ca. 0.0 V vs. the open circuit potential. These spectral changes were interpreted on the basis of the orientation- and distance-dependent IR enhancement and indicate alterations of the inclination angle of the β-strands as crucial molecular events, reflecting an expansion or contraction of the β-barrel pore. These protein structural changes that do not confirm nor exclude the reorientation of the α-helix are either directly induced by the electric field or a consequence of a potential-dependent repulsion or attraction of the bilayer.

Graphical abstract: Voltage-dependent structural changes of the membrane-bound anion channel hVDAC1 probed by SEIRA and electrochemical impedance spectroscopy

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Supplementary files

Article information


Submitted
13 Jan 2014
Accepted
31 Mar 2014
First published
03 Apr 2014

Phys. Chem. Chem. Phys., 2014,16, 9546-9555
Article type
Paper
Author version available

Voltage-dependent structural changes of the membrane-bound anion channel hVDAC1 probed by SEIRA and electrochemical impedance spectroscopy

J. Kozuch, C. Weichbrodt, D. Millo, K. Giller, S. Becker, P. Hildebrandt and C. Steinem, Phys. Chem. Chem. Phys., 2014, 16, 9546
DOI: 10.1039/C4CP00167B

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