Two-step purification of outer membrane proteins.
Here, we describe a simple and efficient method for the purification of Escherichia coli outer membrane proteins. We have tested this protocol for the purification of Wza and Osmoporin C (OmpC) proteins. Both proteins were purified to homogeneity, in two steps, by anion exchange and size exclusion chromatography with a final yield of 92.5 mg for the Wza protein and 291.5 mg for the OmpC protein. The purity of the samples was judged by electrophoretic analysis, mass spectrometry, single particle analysis, three-dimensional (3D) crystallisation and X-ray diffraction.
Beis
Konstantinos
K
Centre for Biomolecular Sciences, University of St. Andrews, St. Andrews, Fife KY16 9ST, UK. kbeis@scripps.edu
Whitfield
Chris
C
Booth
Ian
I
Naismith
James H
JH
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000302:isolation & purification*; D018272:Porins / Q000737:chemistry / Q000302:isolation & purification*
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C113724:OmpC protein; D018272:Porins; C404158:Wza protein, E coli