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Biophys J, May 1998, p. 2649-2657, Vol. 74, No. 5

Characterization of the Growth of 2D Protein Crystals on a Lipid Monolayer by Ellipsometry and Rigidity Measurements Coupled to Electron Microscopy

Catherine Vénien-Bryan,* Pierre-François Lenne,# Cécile Zakri,# Anne Renault,# Alain Brisson,§ Jean-François Legrand, and Bruno Berge#

 *Institut de Biologie Structurale Jean-Pierre Ebel (CEA-CNRS), 38027 Grenoble cedex 1, France;  #Laboratoire de Spectrométrie Physique, Université J. Fourier Grenoble I, BP 87, 38402 St Martin d'Hères, France;  §Department of Chemistry/Biophysical Chemistry, University of Groningen, NL-9747 AG Groningen, The Netherlands; and  SI3M (UMR 585 CEA-CNRS-Université J. Fourier), DRFMC, 38054 Grenoble cedex 9, France

We present here some sensitive optical and mechanical experiments for monitoring the process of formation and growth of two-dimensional (2D) crystals of proteins on a lipid monolayer at an air-water interface. The adsorption of proteins on the lipid monolayer was monitored by ellipsometry measurements. An instrument was developed to measure the shear elastic constant (in plane rigidity) of the monolayer. These experiments have been done using cholera toxin B subunit (CTB) and annexin V as model proteins interacting with a monosialoganglioside (GM1) and dioleoylphosphatidylserine (DOPS), respectively. Electron microscopy observations of the protein-lipid layer transferred to grids were systematically used as a control. We found a good correlation between the measured in-plane rigidity of the monolayer and the presence of large crystalline domains observed by electron microscopy grids. Our interpretation of these data is that the crystallization process of proteins on a lipid monolayer passes through at least three successive stages: 1) molecular recognition between protein and lipid-ligand, i.e., adsorption of the protein on the lipid layer; 2) nucleation and growth of crystalline patches whose percolation is detected by the appearance of a non-zero in-plane rigidity; and 3) annealing of the layer producing a slower increase of the lateral or in-plane rigidity.

Biophys J, May 1998, p. 2649-2657, Vol. 74, No. 5
© 1998 by the Biophysical Society   0006-3495/98/05/2649/09  $2.00


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