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Mechanosensitive Channels Activity in a Droplet Interface Bilayer System

Published online by Cambridge University Press:  30 January 2014

Joseph Najem,
Myles Dunlap,
Sergei Sukharev and
Donald J. Leo
Joseph Najem
Affiliation:
Biomolecular Materials and Systems Laboratory, Virginia Tech, Blacksburg, VA 24061, U.S.A.
Myles Dunlap
Affiliation:
Biomolecular Materials and Systems Laboratory, Virginia Tech, Blacksburg, VA 24061, U.S.A.
Sergei Sukharev
Affiliation:
Department of Biology, University of Maryland, College Park, MD 20742, U.S.A.
Donald J. Leo
Affiliation:
College of Engineering, University of Georgia, Athens, GA 30609, U.S.A.
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Abstract

This paper presents the first attempts to study the large conductance mechano-sensitive channel (MscL) activity in an artificial droplet interface bilayer (DIB) system. A novel and simple technique is developed to characterize the behavior of an artificial lipid bilayer interface containing mechano-sensitive (MS) channels. The experimental setup is assembled on an inverted microscope and consists of two micropipettes filled with PEG-DMA hydrogel and containing Ag/AgCl wires, a cylindrical oil reservoir glued on top of a thin acrylic sheet, and a piezoelectric oscillator actuator. By using this technique, dynamic tension can be applied by oscillating axial motion of one droplet, producing deformation of both droplets and area changes of the DIB interface. The tension in the artificial membrane will cause the MS channels to gate, resulting in an increase in the conductance levels of the membrane. The results show that the MS channels are able to gate under an applied dynamic tension. Moreover, it can be concluded that the response of channel activity to mechanical stimuli is voltage-dependent and highly related to the frequency and amplitude of oscillations.

Keywords

biomaterialself-assemblyactivation analysis

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