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dc.contributor.authorNajem, Joseph S.
dc.contributor.authorDunlap, Myles D.
dc.contributor.authorRowe, Ian D.
dc.contributor.authorFreeman, Eric C.
dc.contributor.authorGrant, John W.
dc.contributor.authorSukharev, Sergei
dc.contributor.authorLeo, Donald J.
dc.date.accessioned2019-01-25T15:47:45Z
dc.date.available2019-01-25T15:47:45Z
dc.date.issued2015-09-08
dc.identifier.issn2045-2322
dc.identifier.other13726
dc.identifier.urihttp://hdl.handle.net/10919/86890
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_US
dc.description.sponsorshipAir Force Office of Scientific Research Basic Research Initiative Grant [FA9550-12-1-0464]
dc.format.extent11
dc.format.mimetypeapplication/pdf
dc.language.isoen_US
dc.publisherSpringer Nature
dc.rightsCreative Commons Attribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectprokaryotic mechanosensitive channels
dc.subjectescherichia-coli
dc.subjectgating mechanism
dc.subjectelectric-field
dc.subjection-channel
dc.subjectmembrane
dc.subjectprotein
dc.titleActivation of bacterial channel MscL in mechanically stimulated droplet interface bilayersen_US
dc.typeArticle - Refereed
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.
dc.title.serialScientific Reports
dc.identifier.doihttps://doi.org/10.1038/srep13726
dc.identifier.volume5
dc.type.dcmitypeText
dc.identifier.pmid26348441


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