Zinc transporter 10 (ZnT10)-dependent extrusion of cellular Mn2+ is driven by an active Ca2+-coupled exchange

Abstract

Manganese (Mn²⁺) is extruded from the cell by the zinc transporter 10 (ZnT10). Loss of ZnT10 expression caused by autosomal mutations in the ZnT10 gene leads to hypermanganesemia in multiple organs. Here, combining fluorescent monitoring of cation influx in HEK293-T cells expressing human ZnT10 with molecular modeling of ZnT10 cation selectivity, we show that ZnT10 is exploiting the transmembrane Ca²⁺ inward gradient for active cellular exchange of Mn²⁺ In analyzing ZnT10 activity we used the ability of Fura-2 to spectrally distinguish between Mn²⁺ and Ca²⁺ fluxes. We found that (a) application of Mn²⁺-containing Ca²⁺-free solution to ZnT10-expressing cells triggers an influx of Mn²⁺, (b) reintroduction of Ca²⁺ leads to cellular Mn²⁺ extrusion against an inward Mn²⁺ gradient, and (c) the cellular transport of Mn²⁺ by ZnT10 is coupled to a reciprocal movement of Ca²⁺ Remarkably, replacing a single asparagine residue in ZnT10 (Asp-43) with threonine (ZnT10 N43T) converted the Mn²⁺/Ca²⁺ exchange to an uncoupled channel mode, permeable to both Ca²⁺ and Mn²⁺ The findings in our study identify the first ion transporter that uses the Ca²⁺ gradient for active counter-ion exchange. They highlight a remarkable versatility in metal selectivity and mode of transport controlled by the tetrahedral metal transport site of ZnT proteins.

Keywords: Ca2+ coupled exchange; active transport; anion transport; calcium transport; cation diffusion facilitator; hypermanganesemia; ion transport; manganese; manganese extrusion; membrane transport; solute carrier family 30 member 10 (SLC30A10); zinc; zinc transport; zinc transporter (ZnT).