Architecture of the Mitochondrial Calcium Uniporter

Mitochondria from multiple, eukaryotic clades uptake and buffer large amounts of calcium (Ca(2+)) via an inner membrane transporter called the uniporter. Early studies demonstrated that this transport requires a mitochondrial membrane potential and that the uniporter is itself Ca(2+) activated, and blocked by ruthenium red or Ru360(1). Later, electrophysiological studies demonstrated that the uniporter is an ion channel with remarkably high conductance and selectivity(2). Ca(2+) entry into mitochondria is also known to activate the TCA cycle and appears to be critical for matching ATP production in mitochondria with its cytosolic demand(3). MCU (mitochondrial calcium uniporter) is the pore forming and Ca(2+) conducting subunit of the uniporter, but its primary sequence does not resemble any calcium channel known to date. Here, we report the structure of the core region of MCU, determined using nuclear magnetic resonance (NMR) and electron microscopy (EM). MCU is a homo-oligomer with the second transmembrane helix forming a hydrophilic pore across the membrane. The channel assembly represents a new solution of ion channel architecture and is stabilized by a coiled coil motif protruding in the mitochondrial matrix. The critical DxxE motif forms the pore entrance featuring two carboxylate rings, which appear to be the selectivity filter based on the ring dimensions and functional mutagenesis. To our knowledge, this is one of the largest structures characterized by NMR, which provides a structural blueprint for understanding the function of this channel.