Salting out the ionic selectivity of a wide channel: the asymmetry of OmpF.
Although the crystallographic structure of the bacterial porin OmpF has been known for a decade, the physical mechanisms of its ionic selectivity are still under investigation. We address this issue in a series of experiments with varied pH, salt concentrations, inverted salt gradient, and charged and uncharged lipids. Measuring reversal potential, we show that OmpF selectivity (traditionally regarded as slightly cationic) depends strongly on pH and salt concentration and is conditionally asymmetric, that is, the calculated selectivity is sensitive to the direction of salt concentration gradient. At neutral pH and subdecimolar salt concentrations the channel exhibits nearly ideal cation selectivity (t(G)(+)=0.98+/-0.01). Substituting neutral DPhPC with DPhPS, we demonstrate that the fixed charge of the host lipid has a small but measurable effect on the channel reversal potential. The available structural information allows for a qualitative explanation of our experimental findings. These findings now lead us to re-examine the ionization state of 102 titratable sites of the OmpF channel. Using standard methods of continuum electrostatics tailored to our particular purpose, we find the charge distribution in the channel as a function of solution acidity and relate the pH-dependent asymmetry in channel selectivity to the pH-dependent asymmetry in charge distribution. In an attempt to find a simple phenomenological description of our results, we also discuss different macroscopic models of electrodiffusion through large channels.
Departamento de Ciencias Experimentales, Universidad Jaume I, Castellón, Spain.
Antonio
A
Alcaraz
Ekaterina M
EM
Nestorovich
Marcel
M
Aguilella-Arzo
Vicente M
VM
Aguilella
Sergey M
SM
Bezrukov
D016880:Anisotropy; D003198:Computer Simulation; D006863:Hydrogen-Ion Concentration; D015640:Ion Channel Gating; D007477:Ions; D008051:Lipid Bilayers / Q000737:chemistry*; D008560:Membrane Fluidity; D008564:Membrane Potentials; D008567:Membranes, Artificial; D008956:Models, Chemical; D008958:Models, Molecular; D010713:Phosphatidylcholines / Q000737:chemistry*; D012492:Salts / Q000737:chemistry*; D055672:Static Electricity
D007477:Ions; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D012492:Salts; C021016:1,2-diphytanoylphosphatidylcholine