Activation of bacterial channel MscL in mechanically stimulated droplet interface bilayers

Scientific Reports
Joseph S NajemDonald J Leo

Abstract

MscL, 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...Continue Reading

References

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Citations

Aug 3, 2017·Molecular BioSystems·Michael J BoothHagan Bayley
Mar 15, 2019·Biomicrofluidics·Elio J ChallitaEric C Freeman
Aug 2, 2017·Scientific Reports·Masayuki Iwamoto, Shigetoshi Oiki
Apr 25, 2018·Scientific Reports·Elio J ChallitaEric C Freeman
Mar 27, 2020·Frontiers in Bioengineering and Biotechnology·Ignacio Poblete-CastroJosé Manuel Borrero-de Acuña
Feb 4, 2021·Journal of the Royal Society, Interface·Y HuangG G Fuller
Mar 10, 2021·Langmuir : the ACS Journal of Surfaces and Colloids·Michelle M Makhoul-Mansour, Eric C Freeman
Feb 23, 2021·Chembiochem : a European Journal of Chemical Biology·Tsoi IpYuval Elani
Apr 8, 2021·Langmuir : the ACS Journal of Surfaces and Colloids·Megan WoodSunghee Lee
Jun 12, 2021·Communications Chemistry·Stuart HaylockOscar Ces
Apr 14, 2021·Bioinspiration & Biomimetics·Michelle M Makhoul-MansourEric C Freeman
Oct 19, 2017·Langmuir : the ACS Journal of Surfaces and Colloids·Michelle Makhoul-MansourEric C Freeman
May 20, 2020·The Journal of Physical Chemistry. B·William T McClinticC Patrick Collier
May 2, 2018·Langmuir : the ACS Journal of Surfaces and Colloids·Peter J BeltramoJan Vermant
Aug 28, 2021·International Journal of Molecular Sciences·Miklós LengyelGábor Czirják

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Methods Mentioned

BETA
chemical modifications
PCR

Software Mentioned

MscL

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