Study of Selectivity and Permeation In Voltage-Gated Ion Channels

TL;DR

This study investigates ion selectivity and permeation in voltage-gated ion channels using reduced models, simulations, and experiments, revealing key determinants of selectivity and the effects of sialic acid on bacterial channels.

Contribution

It introduces a simplified model for calcium channel selectivity, explores the role of side chain flexibility, and characterizes NanC channel behavior and sialic acid specificity in E. coli.

Findings

Charge density in the selectivity filter is the primary determinant of calcium channel selectivity.

Flexibility of side chains has a secondary effect on calcium channel selectivity.

Sialic acid enhances NanC conductance and facilitates translocation in E. coli.

Abstract

Ion channels are membrane proteins responsible for an enormous range of biological functions. Ion selectivity and permeation are based on simple laws of physics and chemistry. Ion channels are therefore ideal candidates for physical investigation. A reduced model generates the selectivity of voltage-gated L-type calcium channel under a wide range of ionic conditions using only two parameters with unchanging values. The reasons behind the success of this reduced model are investigated. Monte Carlo simulations are performed investigating the role of flexibility of the side chains in the selectivity of calcium channels under a wide range of ionic conditions. Results suggest that the exact location and mobility of oxygen ions have little effect on the selectivity behavior of calcium channels. The first order determinant of selectivity in calcium channels is the density of charge in the selectivity filter. Flexibility seems a second order determinant. Single channel planar lipid bilayer experiments are performed to determine the selectivity, permeation, and sialic acid specificity of a bacterial outer-membrane channel NanC of Ecoli. NanC exhibits a large unit conductance varying with the applied voltage polarity, anion selectivity, and voltage gating. Unitary conductance of NanC decreases significantly in presence of the buffer HEPES. Alternate buffers are tested. Results suggest that the sialic acid specificity of NanC should be performed in low concentration salt solutions without pH buffers adjusted to neutral pH 7.0. Neu5Ac changes the gating and considerably increases the ionic conductance of NanC. The effect of Neu5Ac on the unitary current through NanC saturates at higher Neu5Ac concentrations. Interestingly, Neu5Ac reduces the ionic conductance of OmpF: frequent, long closures are seen. Thus, in E.coli sialic acid translocation is specifically facilitated by NanC, and not by OmpF.