Publication
553
Langmuir, 2002,
18 (8), 3263 -3272
DOI: 10.1021/la011585t S0743-7463(01)01585-2 |
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Formation
of Tethered and Streptavidin-Supported Lipid
Bilayers on a Microporous Electrode for the
Reconstitution of Membranes of Large Surface
Area
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Vanessa
Proux-Delrouyre, Jean-Marc Laval,, Jacques Moiroux and
Christian Bourdillon
Laboratoire de Technologie Enzymatique,
UMR du CNRS No. 6022, Université de Technologie
de Compiègne, B.P. 20529, 60205 Compiègne
Cedex, France, and Laboratoire d'Electrochimie Moléculaire,
UMR du CNRS N° 7591, Université Paris
7-Denis Diderot, 75251 Paris Cedex 05, France
A new approach for the self-assembly of supported
and tethered lipid membranes of large surface area
is proposed. The template is a microporous electrode
made by anodic etching of aluminum and covered with
a monolayer of streptavidin. We show that spontaneous
fusion of biotinylated lipid vesicles on the affinity
layer is a slow process despite abundant accumulation
of lipid material at the template surface. To increase
dramatically the efficiency of the self-assembly,
fast fusion is provoked with the help of a fusogen
solution of poly(ethylene glycol). The extent of
fusion is assessed by electrochemical monitoring
of the long-range lateral mobility of ubiquinone
(coenzyme Q10) in the supported bilayer.
Finally, the geometrical characterization of the
honeycomb structure at key steps of the self-assembly
procedure is performed by electrochemical measurement
of the porosity. As expected, the formation of the
supported bilayer causes a decrease in the apparent
inner diameter of the pores. It is expected that
the type of supported lipid membrane built according
to the present approach can be adequate for the incorporation
of transmembrane proteins in structures that would
mimic the membrane stacking found in chloroplasts
or mitochondria. |