Elucidating the H⁺ coupled Zn²⁺ transport mechanism of ZIP4; implications in acrodermatitis enteropathica

Cellular Zn²⁺ homeostasis is tightly regulated and primarily mediated by designated Zn²⁺ transport proteins, namely zinc transporters (ZnTs; SLC30) that shuttle Zn²⁺ efflux, and ZRT-IRT-like proteins (ZIPs; SLC39) that mediate Zn²⁺ influx. While the functional determinants of ZnT-mediated Zn²⁺ efflux are elucidated, those of ZIP transporters are lesser understood. Previous work has suggested three distinct molecular mechanisms: (I) HCO3⁻ or (II) H⁺ coupled Zn²⁺ transport, or (III) a pH regulated electrodiffusional mode of transport. Here, using live-cell fluorescent imaging of Zn²⁺ and H⁺, in cells expressing ZIP4, we set out to interrogate its function. Intracellular pH changes or the presence of HCO3⁻ failed to induce Zn²⁺ influx. In contrast, extracellular acidification stimulated ZIP4 dependent Zn²⁺ uptake. Furthermore, Zn²⁺ uptake was coupled to enhanced H⁺ influx in cells expressing ZIP4, thus indicating that ZIP4 is not acting as a pH regulated channel but rather as an H⁺ powered Zn²⁺ co-transporter. We further illustrate how this functional mechanism is affected by genetic variants in SLC39A4 that in turn lead to Acrodermatitis enteropathica, a rare condition of Zn²⁺ deficiency.