title: High-Throughput Simulations Reveal Membrane-Mediated Effects of Alcohols on MscL Gating.
abstract: The mechanosensitive channels of large conductance (MscL) are bacterial membrane proteins that serve as last resort emergency release valves in case of severe osmotic downshock. Sensing bilayer tension, MscL channels are sensitive to changes in the bilayer environment and are, therefore, an ideal test case for exploring membrane protein coupling. Here, we use high-throughput coarse-grained molecular dynamics simulations to characterize MscL gating kinetics in different bilayer environments under the influence of alcohols. We performed over five hundred simulations to obtain sufficient statistics to reveal the subtle effects of changes in the membrane environment on MscL gating. MscL opening times were found to increase with the addition of the straight-chain alcohols ethanol, octanol, and to some extent dodecanol but not with hexadecanol. Increasing concentration of octanol increased the impeding effect, but only up to 10-20 mol %. Our in silico predictions were experimentally confirmed using reconstituted MscL in a liposomal fluorescent efflux assay. Our combined data reveal that the effect of alcohols on MscL gating arises not through specific binding sites but through a combination of the alcohol-induced changes to a number of bilayer properties and their alteration of the MscL-bilayer interface. Our work provides a key example of how extensive molecular simulations can be used to predict the functional modification of membrane proteins by subtle changes in their bilayer environment.
authors.affiliation: Groningen Biomolecular Science and Biotechnology Institute, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.|Zernike Institute for Advanced Materials, University of Groningen , Nijenborgh 4, 9747 AG, Groningen, The Netherlands.
authors.forename: Manuel N
authors.initials: MN
authors.lastname: Melo
authors.affiliation: Groningen Biomolecular Science and Biotechnology Institute, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.|Zernike Institute for Advanced Materials, University of Groningen , Nijenborgh 4, 9747 AG, Groningen, The Netherlands.
authors.forename: Clément
authors.initials: C
authors.lastname: Arnarez
authors.affiliation: Groningen Biomolecular Science and Biotechnology Institute, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.|Zernike Institute for Advanced Materials, University of Groningen , Nijenborgh 4, 9747 AG, Groningen, The Netherlands.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Sikkema
authors.affiliation: Groningen Institute for Evolutionary Life Sciences, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.
authors.forename: Neeraj
authors.initials: N
authors.lastname: Kumar
authors.affiliation: Groningen Biomolecular Science and Biotechnology Institute, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.
authors.forename: Martin
authors.initials: M
authors.lastname: Walko
authors.affiliation: Groningen Biomolecular Science and Biotechnology Institute, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.|Zernike Institute for Advanced Materials, University of Groningen , Nijenborgh 4, 9747 AG, Groningen, The Netherlands.
authors.forename: Herman J C
authors.initials: HJ
authors.lastname: Berendsen
authors.affiliation: Department of Neuroscience, University Medical Center Groningen, University of Groningen , Antonius Deusinglaan 1, 99713 AV, Groningen, The Netherlands.
authors.forename: Armagan
authors.initials: A
authors.lastname: Kocer
authors.affiliation: Groningen Biomolecular Science and Biotechnology Institute, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.|Zernike Institute for Advanced Materials, University of Groningen , Nijenborgh 4, 9747 AG, Groningen, The Netherlands.
authors.forename: Siewert J
authors.initials: SJ
authors.lastname: Marrink
authors.affiliation: Groningen Biomolecular Science and Biotechnology Institute, University of Groningen , Nijenborgh 7, 9747 AG, Groningen, The Netherlands.|Zernike Institute for Advanced Materials, University of Groningen , Nijenborgh 4, 9747 AG, Groningen, The Netherlands.|Biosciences and Biotechnology Division, Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory , Livermore, California.
authors.forename: Helgi I
authors.identifier: 0000-0002-7613-9143
authors.initials: HI
authors.lastname: Ingólfsson
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000502:physiology; D002462:Cell Membrane / Q000737:chemistry* / Q000502:physiology; D000431:Ethanol; D015640:Ion Channel Gating / Q000502:physiology; D007473:Ion Channels / Q000502:physiology*; D055595:Mechanical Phenomena; D056004:Molecular Dynamics Simulation; D000442:Octanols
chemical_list: D001426:Bacterial Proteins; D007473:Ion Channels; D000442:Octanols; D000431:Ethanol