(A–B) Low [Ca2+] structures. (A) TcMCU-EMRE-MICU1-MICU2 holocomplex structure reported here. (B) Holocomplex containing human MCU, EMRE, MICU1, and MICU2 subunits (PDB # GWDN). An ordered loop of MICU1 (amino acids 258–274) is indicated, as are the C-helices of MICU1 and MICU2, which are also ordered in that structure. (C) Structure of a holocomplex comprising human MCU, EMRE, MICU1, and MICU2 that was determined at high (2 mM) [Ca2+] (PDB # GWDO). A dimer of channels present in the cryo-EM sample is formed by interactions between the NTDs. For each channel, a Ca2+-bound MICU1-MICU2 regulatory complex is located adjacent to the pore and no longer occludes it.