Hierarchical approach to predicting permeation in ion channels

A hierarchical computational strategy combining molecular modeling, electro statics calculations, molecular dynamics, and Brownian dynamics simulations is developed and implemented to compute electrophysiologically measurable properties of the KcsA potassium channel. Models for a series of channels w ith different pore sizes are developed from the known x-ray structure, usin g Insights into the gating conformational changes as suggested by a variety of published experiments. Information on the pH dependence of the channel gating is incorporated into the calculation of potential profiles for K+ io ns inside the channel, which are then combined with K+ ion mobilities insid e the channel, as computed by molecular dynamics simulations, to provide in puts into Brownian dynamics simulations for computing ion fluxes. The open model structure has a conductance of similar to 110 pS under symmetric 250 mM K+ conditions, in reasonable agreement with experiments for the largest conducting substate. The dimensions of this channel are consistent with ele ctrophysiologically determined size dependence of quaternary ammonium ion b locking from the intracellular end of this channel as well as with direct s tructural evidence that tetrabutylammonium ions can enter into the interior cavity of the channel. Realistic values of Ussing flux ratio exponents, di stribution of ions within the channel, and shapes of the current-voltage an d current-concentration curves are obtained. The Brownian dynamics calculat ions suggest passage of ions through the selectivity filter proceeds by a " knock-off" mechanism involving three ions, as has been previously inferred from functional and structural studies of barium ion blocking. These result s suggest that the present calculations capture the essential nature of Kion permeation in the KcsA channel and provide a proof-of-concept for the i ntegrated microscopic/mesoscopic multitiered approach for predicting ion ch annel function from structure, which can be applied to other channel struct ures.