Two-dimensional crystallization of a bacterial surface protein on lipid vesicles under controlled conditions

Biophys J. 1992 Jan;61(1):172-88. doi: 10.1016/S0006-3495(92)81825-8.

Abstract

The solubilized surface protein of the Gram-negative bacterium Comamonas acidovorans was reconstituted on lipid vesicles by means of controlled dialysis. To this end, a multichamber dialysis apparatus was built which allows one to control the temperature and the dialysis rate, to apply various temperatures or buffer systems and sample conditions in a single experiment, and to monitor the turbidity of the sample by means of light scattering. The reconstitution conditions were optimized such that the surface protein formed two-dimensional crystals suitable for electron crystallography. The recrystallized surface protein arrays gave a resolution of approximately 1.3 nm in projection after correlation averaging of negatively stained preparations. The surface protein assembled into partially self-contained two-dimensional crystals which possess a strong shape-determining effect and formed cylinders and various cone-shaped vesicles. The development of the various vesicle forms is described in a model.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Outer Membrane Proteins / isolation & purification
  • Bacterial Outer Membrane Proteins / ultrastructure*
  • Crystallization
  • Dimyristoylphosphatidylcholine*
  • Light
  • Liposomes*
  • Microscopy, Electron
  • Models, Structural
  • Protein Conformation
  • Pseudomonas
  • Scattering, Radiation
  • Thermodynamics

Substances

  • Bacterial Outer Membrane Proteins
  • Liposomes
  • Dimyristoylphosphatidylcholine