title: Activation of the mechanosensitive ion channel MscL by mechanical stimulation of supported Droplet-Hydrogel bilayers.
abstract: The droplet on hydrogel bilayer (DHB) is a novel platform for investigating the function of ion channels. Advantages of this setup include tight control of all bilayer components, which is compelling for the investigation of mechanosensitive (MS) ion channels, since they are highly sensitive to their lipid environment. However, the activation of MS ion channels in planar supported lipid bilayers, such as the DHB, has not yet been established. Here we present the activation of the large conductance MS channel of E. coli, (MscL), in DHBs. By selectively stretching the droplet monolayer with nanolitre injections of buffer, we induced quantifiable DHB tension, which could be related to channel activity. The MscL activity response revealed that the droplet monolayer tension equilibrated over time, likely by insertion of lipid from solution. Our study thus establishes a method to controllably activate MS channels in DHBs and thereby advances studies of MS channels in this novel platform.
authors.affiliation: The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW 2010, Australia.
authors.forename: Kadla R
authors.initials: KR
authors.lastname: Rosholm
authors.affiliation: School of Medical Sciences, University of New South Wales, Kensington, NSW 2052, Australia.
authors.forename: Matthew A B
authors.initials: MA
authors.lastname: Baker
authors.affiliation: The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW 2010, Australia.
authors.forename: Pietro
authors.initials: P
authors.lastname: Ridone
authors.affiliation: The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW 2010, Australia.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Nakayama
authors.affiliation: The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW 2010, Australia.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Rohde
authors.affiliation: Department of Cell Physiology and Molecular Biophysics, Center for Membrane Protein Research, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
authors.forename: Luis G
authors.initials: LG
authors.lastname: Cuello
authors.affiliation: The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW 2010, Australia.|School of Medical Sciences, University of New South Wales, Kensington, NSW 2052, Australia.
authors.forename: Lawrence K
authors.initials: LK
authors.lastname: Lee
authors.affiliation: The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW 2010, Australia.|St Vincent's Clinical School, Darlinghurst, NSW 2010, Australia.
authors.forename: Boris
authors.initials: B
authors.lastname: Martinac
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D020100:Hydrogels; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism*; D008051:Lipid Bilayers / Q000378:metabolism; D066292:Lipid Droplets / Q000378:metabolism; D040542:Mechanotransduction, Cellular; D009154:Mutation
chemical_list: D029968:Escherichia coli Proteins; D020100:Hydrogels; D007473:Ion Channels; D008051:Lipid Bilayers; C086596:MscL protein, E coli