- Powered by the steep mitochondrial membrane potential Ca 2+ permeates into the mitochondria via the Ca 2+ uniporter and is then extruded by a mitochondrial Na + /Ca 2+ exchanger. This mitochondrial Ca 2+ shuttling regulates the rate of ATP production and participates in cellular Ca 2+ signaling. Despite the fact that the exchanger was functionally identified 40 years ago its molecular identity remained a mystery. Early studies on isolated mitochondria and intact cells characterized the functional properties of a mitochondrial Na + /Ca 2+ exchanger, and showed that it possess unique functional fingerprints such as Li + /Ca 2+ exchange and that it is displaying selective sensitivity to inhibitors. Purification of mitochondria proteins combined with functional reconstitution led to the isolation of a polypeptide candidate of the exchanger but failed to molecularly identify it. A turning point in the search for the exchanger molecule came with the recent cloning of the last member of the Na + /Ca 2+ exchanger superfamily termed NCLX (Na + /Ca 2+ /Li + exchanger). NCLX is localized in the inner mitochondria membrane and its expression is linked to mitochondria Na + /Ca 2+ exchange matching the functional fingerprints of the putative mitochondrial Na + /Ca 2+ exchanger. Thus NCLX emerges as the long sought mitochondria Na + /Ca 2+ exchanger and provide a critical molecular handle to study mitochondrial Ca 2+ signaling and transport. Here we summarize some of the main topics related to the molecular properties of the Na + /Ca 2+ exchanger, beginning with the early days of its functional identification, its kinetic properties and regulation, and culminating in its molecular identification.