Two-dimensional crystallisation of soluble protein complexes.
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.
Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, 1, rue Laurent Fries, BP163, 67404 Illkirch-Graffentaden, France.
Patrick
P
Schultz
Corinne
C
Crucifix
Luc
L
Lebeau
D003460:Crystallization / Q000379:methods*; D004247:DNA / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000648:ultrastructure; D008055:Lipids / Q000737:chemistry; D008854:Microscopy, Electron; D011506:Proteins / Q000737:chemistry* / Q000648:ultrastructure; D012995:Solubility; D019809:Streptavidin / Q000737:chemistry / Q000648:ultrastructure
D008055:Lipids; D011506:Proteins; D004247:DNA; D019809:Streptavidin; D012321:DNA-Directed RNA Polymerases