Human genetic variants in SLC39A8 impact uptake and steady-state metal levels within the cell

<jats:p>The human<jats:italic>SLC39A8</jats:italic>(<jats:italic>hSLC39A8</jats:italic>) gene encodes a plasma membrane protein SLC39A8 (ZIP8) that mediates the specific uptake of the metals Cd<jats:sup>2+</jats:sup>, Mn<jats:sup>2+</jats:sup>, Zn<jats:sup>2+</jats:sup>, Fe<jats:sup>2+</jats:sup>, Co<jats:sup>2+</jats:sup>, and Se<jats:sup>4+</jats:sup>. Pathogenic variants within<jats:italic>hSLC39A8</jats:italic>are associated with congenital disorder of glycosylation type 2 (CDG type II) or Leigh-like syndrome. However, numerous mutations of uncertain significance are also linked to different conditions or benign traits. Our study characterized 21<jats:italic>hSLC39A8</jats:italic>variants and measured their impact on protein localization and intracellular levels of Cd<jats:sup>2+</jats:sup>, Zn<jats:sup>2+</jats:sup>, and Mn<jats:sup>2+</jats:sup>. We identified four variants that disrupt protein expression, five variants with high retention in the endoplasmic reticulum, and 12 variants with localization to the plasma membrane. From the 12 variants with plasma membrane localization, we identified three with complete loss of detectable ion uptake by the cell and five with differential uptake between metal ions. Further in silico analysis on protein stability identified variants that may affect the stability of homodimer interfaces. This study elucidates the variety of effects of<jats:italic>hSLC39A8</jats:italic>variants on ZIP8 and on diseases involving disrupted metal ion homeostasis.</jats:p>