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bioRxiv · 10.1101/2022.04.01.486690

A co-operative knock-on mechanism underpins Ca2+-selective cation permeation in TRPV channels

Ives, C. M.·Thomson, N. J.·Zachariae, U.
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Abstract

The selective exchange of ions across cellular membranes is a vital biological process. Ca2+-mediated signalling is implicated in a broad array of physiological processes in cells, whilst elevated intracellular concentrations of Ca2+ are cytotoxic. Due to the significance of this cation, strict Ca2+ concentration gradients are maintained across the plasma and organelle membranes. Therefore, Ca2+ signalling relies on permeation through selective ion channels that control the flux of Ca2+ ions. A key family of Ca2+-permeable membrane channels are the polymodal signal-detecting Transient Receptor Potential (TRP) ion channels. TRP channels are activated by a wide variety of cues including temperature, small molecules, transmembrane voltage and mechanical stimuli. Whilst most members of this family permeate a broad range of cations non-selectively, TRPV5 and TRPV6 are unique due to their strong Ca2+-selectivity. Here, we address the question of how some members of the TRPV subfamily show a high degree of Ca2+-selectivity whilst others conduct a wider spectrum of cations. We present results from all-atom molecular dynamics simulations of ion permeation through two Ca2+-selective and two non-selective TRPV channels. Using a new method to quantify permeation co-operativity based on mutual information, we show that Ca2+-selective TRPV channel permeation occurs by a three binding site knock-on mechanism, whereas a two binding site knock-on mechanism is observed in non-selective TRPV channels. Each of the ion binding sites involved displayed greater affinity for Ca2+ over Na+. As such, our results suggest that coupling to an extra binding site in the Ca2+-selective TRPV channels underpins their increased selectivity for Ca2+ over Na+ ions. Furthermore, analysis of all available TRPV channel structures shows that the selectivity filter entrance region is wider for the non-selective TRPV channels, slightly destabilising ion binding at this site, which is likely to underlie mechanistic decoupling.

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Ives, C. M., Thomson, N. J., Zachariae, U.. 2022-04-03. A co-operative knock-on mechanism underpins Ca2+-selective cation permeation in TRPV channels. https://doi.org/10.1101/2022.04.01.486690

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