Biophys J, July 2000, p. 496-500, Vol. 79, No. 1

Synchrotron Radiation Diffraction from Two-Dimensional Protein Crystals at the Air/Water Interface

Pierre-François Lenne,^* Bruno Berge,^* Anne Renault,^* Cécile Zakri,^* Catherine Vénien-Bryan, Sébastien Courty, Fabrice Balavoine, W. Bergsma-Schutter,^§ Alain Brisson,^§ Gerhard Grübel,^¶ Nathalie Boudet,^¶ Oleg Konovalov,^¶ and Jean-François Legrand

 ^*Laboratoire de Spectrométrie Physique, UMR Centre National de la Recherche Scientifique-Université J. Fourier, 38041 Grenoble, France;  Institut de Biologie Structurale, CEA-Centre National de la Recherche Scientifique, 38027 Grenoble, France;  Département de Biologie Cellulaire et Moléculaire, CEA Saclay, 91191 Gif sur Yvette, France;  ^§Department of Biophysical Chemistry, University of Groningen, 9747 AG Groningen, the Netherlands;  ^¶European Synchrotron Radiation Facility, 38043 Grenoble, France; and  UMR SPrAM, CEA-Centre National de la Recherche Scientifique-Université J. Fourier, Département de Recherches Fondamentales sur la Matière Condensée, CEA Grenoble, 38054 Grenoble, France

Protein structure determination by classical x-ray crystallography requires three-dimensional crystals that are difficult to obtain for most proteins and especially for membrane proteins. An alternative is to grow two-dimensional (2D) crystals by adsorbing proteins to ligand-lipid monolayers at the surface of water. This confined geometry requires only small amounts of material and offers numerous advantages: self-assembly and ordering over micrometer scales is easier to obtain in two dimensions; although fully hydrated, the crystals are sufficiently rigid to be investigated by various techniques, such as electron crystallography or micromechanical measurements. Here we report structural studies, using grazing incidence synchrotron x-ray diffraction, of three different 2D protein crystals at the air-water interface, namely streptavidine, annexin V, and the transcription factor HupR. Using a set-up of high angular resolution, we observe narrow Bragg reflections showing long-range crystalline order in two dimensions. In the case of streptavidin the angular range of the observed diffraction corresponds to a resolution of 10 Å in plane and 14 Å normal to the plane. We show that this approach is complementary to electron crystallography but without the need for transfer of the monolayer onto a grid. Moreover, as the 2D crystals are accessible from the buffer solution, the formation and structure of protein complexes can be investigated in situ.

Biophys J, July 2000, p. 496-500, Vol. 79, No. 1
© 2000 by the Biophysical Society   0006-3495/00/07/496/05  $2.00

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