Increased salt concentration promotes competitive block of OmpF channel by protons.
Porins are channel-forming proteins that are located in the outer membranes (OM) of Gram-negative bacteria and allow the influx of hydrophilic nutrients and the extrusion of waste products. The fine regulation of the ion transport through these wide channels could play an important role in the survival of the bacteria in acidic media. We investigate here the mechanism responsible for the pH sensitivity of the trimeric porin OmpF, of Escherichia coli. Planar lipid bilayer electrophysiology and site-directed mutagenesis were used to study the effect of pH on the ion conductive properties of the OmpF channel in its fully open, "nongated" conformation. At low pH we observe a large drop in the OmpF open channel conductance that is accompanied by a substantial increase of the current noise. These channel features are strongly dependent on the salt concentration and we propose that they are originated by competitive binding of cations and protons occurring in the narrow central constriction of the channel. This subtle mechanism reveals to be capital for the channel function because it not only drives the channel sensitivity to pH but is also indispensable for the particularly efficient permeation mechanism of the channel at physiological conditions (~neutral pH).
Department of Physics, University Jaume, Castellon, Spain.
Antonio
A
Alcaraz
María
M
Queralt-Martín
Elena
E
García-Giménez
Vicente M
VM
Aguilella
D004926:Escherichia coli / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D018272:Porins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D011188:Potassium / Q000737:chemistry; D011522:Protons; D012492:Salts / Q000737:chemistry*
C109237:OmpF protein; D018272:Porins; D011522:Protons; D012492:Salts; D011188:Potassium