Cryo-EM structure of the mitochondrial calcium uniporter

The mitochondrial calcium uniporter is a highly selective calcium channel localized to the inner mitochondrial membrane. Here, we describe the structure of an MCU ortholog from the fungus Neosartorya fischeri (NfMCU) determined to 3.8 Å resolution by phase-plate cryo-electron microscopy. The channel is a homotetramer with two-fold symmetry in its amino terminal domain (NTD) that adopts a similar structure to that of human MCU. The NTD assembles as a dimer of dimer to form a tetrameric ring that connects to the transmembrane domain through an elongated coiled-coil domain. The ion conducting pore domain maintains four-fold symmetry with the selectivity filter positioned at the start of the pore forming TM2 helix. The aspartate and glutamate sidechains of the conserved DIME motif are oriented toward the central axis and separated by one helical turn. Thus, the structure of NfMCU offers new insights into channel assembly, selective calcium permeation, and inhibitor binding.