Free-standing lipid films stabilized by Annexin-A5.
Free-standing lipid bilayers in nano- and micro-pores are interesting membrane models and attractive for biotechnological applications. We describe here the controlled preparation of proteo-lipid mono- and bilayers using the Langmuir-Schaefer transfer or Langmuir-Blodgett technique, respectively on hydrophobic and hydrophilic surfaces. We demonstrate the formation of suspended proteo-lipid layers by Transmission Electron Microscopy (TEM) and in situ Atomic Force Microscopy (AFM) imaging. Using Annexin-A5 as a membrane-associated protein, continuous proteo-lipid mono- and bilayers were formed, which span pore arrays over areas of several square-micrometers. The 2D organization of proteins associated to lipid monolayer is well preserved during the transfer process and the protein association is Ca(2+)-dependent and therefore reversible. The simple formation and reliable transfer of stabilized free-standing lipid films is a first crucial step to create biomimetic membranes for biotechnological applications and membrane protein research.
Université de Lyon 1, CNRS, INSA de Lyon, CPE Lyon, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, UMR5246, F-69622, Villeurbanne, France; Laboratoire Chimie & Biologie des Membranes & Nanonoobjets, Université Bordeaux, CBMN, UMR 5248, Allée G. Saint Hilaire, F-33600 Pessac, France. Electronic address: Anne.Simon@univ-lyon1.fr.
Anne
A
Simon
Céline
C
Gounou
Sisareuth
S
Tan
Louis
L
Tiefenauer
Marco
M
Di Berardino
Alain R
AR
Brisson
D017304:Annexin A5 / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008051:Lipid Bilayers / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission
D017304:Annexin A5; D008051:Lipid Bilayers
2D crystal; AFM; Annexin-A5; AnxA5; Atomic Force Microscopy; Free-standing; Lipid bilayer; Membrane protein; TEM; Transmission Electron Microscopy