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Activation of bacterial channel MscL in mechanically stimulated droplet interface bilayers

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dc.contributor.authorNajem, Joseph S.en
dc.contributor.authorDunlap, Myles D.en
dc.contributor.authorRowe, Ian D.en
dc.contributor.authorFreeman, Eric C.en
dc.contributor.authorGrant, John Wallaceen
dc.contributor.authorSukharev, Sergeien
dc.contributor.authorLeo, Donald J.en
dc.contributor.departmentMechanical Engineeringen
dc.contributor.departmentBiomedical Engineering and Mechanicsen
dc.contributor.departmentSchool of Biomedical Engineering and Sciencesen
dc.date.accessioned2019-01-25T15:47:45Zen
dc.date.available2019-01-25T15:47:45Zen
dc.date.issued2015-09-08en
dc.description.abstractMscL, a stretch-activated channel, saves bacteria experiencing hypo-osmotic shocks from lysis. Its high conductance and controllable activation makes it a strong candidate to serve as a transducer in stimuli-responsive biomolecular materials. Droplet interface bilayers (DIBs), flexible insulating scaffolds for such materials, can be used as a new platform for incorporation and activation of MscL. Here, we report the first reconstitution and activation of the low-threshold V23T mutant of MscL in a DIB as a response to axial compressions of the droplets. Gating occurs near maximum compression of both droplets where tension in the membrane is maximal. The observed 0.1-3 nS conductance levels correspond to the V23T-MscL sub-conductive and fully open states recorded in native bacterial membranes or liposomes. Geometrical analysis of droplets during compression indicates that both contact angle and total area of the water-oil interfaces contribute to the generation of tension in the bilayer. The measured expansion of the interfaces by 2.5% is predicted to generate a 4-6 mN/m tension in the bilayer, just sufficient for gating. This work clarifies the principles of interconversion between bulk and surface forces in the DIB, facilitates the measurements of fundamental membrane properties, and improves our understanding of MscL response to membrane tension.en
dc.description.notesWe would like to acknowledge A. Yasmann for the technical assistance and the financial support provided by the Air Force Office of Scientific Research Basic Research Initiative Grant FA9550-12-1-0464.en
dc.description.sponsorshipAir Force Office of Scientific Research Basic Research Initiative Grant [FA9550-12-1-0464]en
dc.format.extent11en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1038/srep13726en
dc.identifier.issn2045-2322en
dc.identifier.other13726en
dc.identifier.pmid26348441en
dc.identifier.urihttp://hdl.handle.net/10919/86890en
dc.identifier.volume5en
dc.language.isoenen
dc.publisherSpringer Natureen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectprokaryotic mechanosensitive channelsen
dc.subjectescherichia-colien
dc.subjectgating mechanismen
dc.subjectelectric-fielden
dc.subjection-channelen
dc.subjectmembraneen
dc.subjectproteinen
dc.titleActivation of bacterial channel MscL in mechanically stimulated droplet interface bilayersen
dc.title.serialScientific Reportsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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