Two-dimensional crystallisation of soluble protein complexes

Patrick Schultz (1, 2) , Corinne Crucifix (2, 1) , Luc Lebeau (3)

1 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire
2 IGBMC - Institut de génétique et biologie moléculaire et cellulaire
3 CAMB - Conception et application de molécules bioactives

This method aims at providing structural information on protein or nucleoprotein complexes by high-resolution electron microscopy. The objective is to promote the self-assembly of the macromolecules into two-dimensional crystals in order to use electron crystallography methods. When combined with observations in the frozen hydrated states and dedicated image processing software these methods can provide detailed 3-D models of the complex. The 2-D crystals of soluble nucleoprotein complexes are formed on lipid monolayers spread at the air-water interface. The macromolecule of interest is targeted to the monolayer by either electrostatic or ligand-induced interactions with the hydrophilic head group of the lipid. Upon interaction with the lipids, the nucleoprotein complex is concentrated at the vicinity of the lipid layer whose in-plane mobility facilitates the contacts between macromolecules and the formation of ordered arrays.

Domaines: Biochimie, Biologie Moléculaire

Patrick Schultz, Corinne Crucifix, Luc Lebeau. Two-dimensional crystallisation of soluble protein complexes. Methods in Molecular Biology, 2009, Methods in Molecular Biology, 543, pp.353-367. ⟨10.1007/978-1-60327-015-1_22⟩. ⟨hal-04127597⟩