Activity Report 2010 - CNRS

SCIENTIFIC REPORT
A polyelectrolyte multilayer film was thus
developed by the team to provide
sufficient mechanical stability and ion
porosity to the bilayer membrane.
Fig. 6: Polyelectrolyte membrane (in blue)
made of 16 PAH/PSS layers constructed over
3mm diameter holes in the Delrin carrierdevice
(in grey)
Another critical aspect of the project is to
achieve a large density of ion channels in
the bilayer membrane. During the 2008-
2010 period, the team indeed studied
various membrane proteins and
developed a method to incorporate some
of them into large unilamellar vesicles
(Fig. 7).
The activity of the membrane proteins
incorporated in the large unilamellar
vesicles was checked by electrophysiological
methods. It is now
necessary to upscale the method by coincorporating
all the necessary proteins
and by fusing all these large unilamellar
vesicles into a single membrane bilayer
at the top of the polyelectrolyte
multilayer film. The resulting biomimetic
membranes will be tested in a parallel
diffusion chamber apparatus (Fig. 8).
Fig. 8: Top : Patch-clamp recording of porins
incorporated into large unilamellar vesicles by
the method explained in Fig. 7.
Bottom: diffusion chamber apparatus to
measure simultaneously ion currents across 6
different biomimetic membranes
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Fig. 7: Top: general procedure for the
formation or large unilamellar vesicles
containing membrane proteins (VDAV).
Bottom: Purified and concentrated
fluorescently labelled vesicles containing
membrane proteins (scale bar = 100 µm)