X-ray diffraction analysis of matrix porin, an integral membrane protein from Escherichia coli outer membranes

Abstract

An integral membrane protein forming channels across Escherichia coli outer membranes, porin, has been crystallized using a polyethylene glycol or salt generated two-phase system. Monodispersity and homogeneity of proteindetergent complexes were found to be prerequisites for reproducible formation of crystals amenable to X-ray structural analysis. By varying pH, detergent and buffer type, large crystals of three different habits can be obtained, two of which are discussed in this paper. The tetragonal form (space group P4 2; unit cell dimensions, a = b = 155 A ̊ , c = 172 A ̊ ) is suitable for X-ray analysis. Low temperature induces a change of the space group to P4 222, with a single trimer in the asymmetric unit. This crystal form diffracts to a resolution beyond 2.9 Å. The hexagonal crystal form (space group P6 322; unit cell dimensions, a = b = 93 A ̊ , c = 220 A ̊ ) is limited in resolution to 4.5 Å, but reveals a packing arrangement very similar to that in two-dimensional membrane-like crystalline arrays.