Phosphoporin PhoE of Escherichia coli: Pore formation and function are determined by the core sugar moiety of lipopolysaccharide

doi:10.1074/jbc.M201950200

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Papers In Press, published online ahead of print June 28, 2002
J. Biol. Chem, 10.1074/jbc.M201950200
Submitted on February 27, 2002
Revised on June 11, 2002
Accepted on June 27, 2002

Phosphoporin PhoE of Escherichia coli: Pore formation and function are determined by the core sugar moiety of lipopolysaccharide

Sven O. Hagge, Hans de Cock, Thomas Gutsmann, Frank Beckers, Ulrich Seydel, and Andre Wiese

Division of Biophysics, Research Center Borstel, Borstel D-23845

Corresponding Author: awiese{at}fz-borstel.de

The lipid matrix of the outer membrane of Gram-negative bacteria is an asymmetric bilayer composed of a phospholipid inner leaflet and a lipopolysaccharide outer leaflet. Incorporated into this lipid matrix are, among other macromolecules, the porins, which have a sieve-like function for the transport or exclusion of hydrophilic substances. It is known that a reduced amount of porins is found in the outer membrane of rough mutants as compared to wild-type bacteria. This observation was discussed to be due to a reduced number of insertion sites in the former. We performed electrical measurements on reconstituted planar bilayers composed of lipopolysaccharide on the one and a phospholipid mixture on the other side, using lipopolysaccharide from various rough mutant strains of Salmonella enterica serovar Minnesota. We found that pore formation by PhoE trimers added to the phospholipid side of the bilayers increased with increasing length of the lipopolysaccharide core sugar moiety. These results allow to conclude that the length of the sugar moiety of lipopolysaccharide is the parameter governing pore formation and that no particular insertion sites are required. We, furthermore, found that also the voltage gating of the porin channels is strongly dependent on the composition of the lipid matrix.

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