Opening a hydrophobic gate: the nicotinic acetylcholine receptor as an example

TL;DR

This study investigates how much the pore of the nicotinic acetylcholine receptor must expand to open, using computational models to determine the minimal expansion needed for ion permeation without requiring helix rotation.

Contribution

It introduces a simple pore expansion method to quantify the minimal structural change needed to open the nAChR channel, challenging previous models requiring helix rotation.

Findings

A pore radius increase of about 1.5 Å opens the channel.

Pore expansion alone can account for channel opening without helix rotation.

Results align with mutational and structural data.

Abstract

To what extent must a hydrophobic gate expand for the channel to count as open? We address this question using the nicotinic acetylcholine receptor (nAChR) as the exemplar. The nAChR is an integral membrane protein which forms a cation selective channel gated by neurotransmitter binding to its extracellular domain. A hydrophobic gating model has been proposed for the nAChR, whereby the pore is incompletely occluded in the closed state channel, with a narrow hydrophobic central gate region which presents an energetic barrier to ion permeation. The nAChR pore is lined by a parallel bundle of five M2 alpha-helices, with the gate formed by three rings of hydrophobic sidechains (9', 13', and 17' of M2). A number of models have been proposed to describe the nature of the conformational change underlying the closed to open transition of the nAChR. These models involve different degrees of M2 helix displacement, rotation, and/or kinking. In this study, we use a simple pore expansion method (previously used to model opening of potassium channels) to generate a series of progressively wider models of the nAChR transmembrane domain. Continuum electrostatics calculations are used to assess the change in the barrier height of the hydrophobic gate as a function of pore expansion. The results suggest that an increase in radius of Delta r ~ 1.5 angstrom is sufficient to functionally open the pore without, for example, a requirement for rotation of the M2 helices. This is evaluated in the context of current mutational and structural data on the nAChR and its homologues.