UniProt UniProt http://www.uniprot.org fan-2020-3 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Fan M., Zhang J., Tsai CW., Orlando BJ., Rodriguez M., Xu Y., Liao M., Tsai MF., Feng L. Expressed in MCU/EMRE/MICU1 -/- cells Nature 2020 Only protein-protein interactions imex curation imex curation fan-2020-1 human-293s Human 293S cell line Human 293s 293S cell line 3d-em-single electron microscopy 3D single particle reconstruction 3D-EM-single structure determination identification by structure determination identification by structure determination structure determination Fan M., Zhang J., Tsai CW., Orlando BJ., Rodriguez M., Xu Y., Liao M., Tsai MF., Feng L. Nature 2020 Only protein-protein interactions imex curation imex curation fan-2020-2 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column experimental particp experimental participant identification experimental particp Fan M., Zhang J., Tsai CW., Orlando BJ., Rodriguez M., Xu Y., Liao M., Tsai MF., Feng L. Nature 2020 Only protein-protein interactions imex curation imex curation micu2_human Calcium uptake protein 2, mitochondrial MICU2 EF-hand domain-containing family member A1 EFHA1 protein protein human Homo sapiens Human MAAAAGSCARVAAWGGKLRRGLAVSRQAVRSPGPLAAAVAGAALAGAGAAWHHSRVSVAARDGSFTVSAQKNVEHGIIYIGKPSLRKQRFMQFSSLEHEGEYYMTPRDFLFSVMFEQMERKTSVKKLTKKDIEDTLSGIQTAGCGSTFFRDLGDKGLISYTEYLFLLTILTKPHSGFHVAFKMLDTDGNEMIEKREFFKLQKIISKQDDLMTVKTNETGYQEAIVKEPEINTTLQMRFFGKRGQRKLHYKEFRRFMENLQTEIQEMEFLQFSKGLSFMRKEDFAEWLLFFTNTENKDIYWKNVREKLSAGESISLDEFKSFCHFTTHLEDFAIAMQMFSLAHRPVRLAEFKRAVKVATGQELSNNILDTVFKIFDLDGDECLSHEEFLGVLKNRMHRGLWVPQHQSIQEYWKCVKKESIKGVKEVWKQAGKGLF micu1_human Calcium uptake protein 1, mitochondrial MICU1 Calcium-binding atopy-related autoantigen 1 CBARA1 CALC Atopy-related autoantigen CALC protein protein human Homo sapiens Human MFRLNSLSALAELAVGSRWYHGGSQPIQIRRRLMMVAFLGASAVTASTGLLWKRAHAESPPCVDNLKSDIGDKGKNKDEGDVCNHEKKTADLAPHPEEKKKKRSGFRDRKVMEYENRIRAYSTPDKIFRYFATLKVISEPGEAEVFMTPEDFVRSITPNEKQPEHLGLDQYIIKRFDGKKISQEREKFADEGSIFYTLGECGLISFSDYIFLTTVLSTPQRNFEIAFKMFDLNGDGEVDMEEFEQVQSIIRSQTSMGMRHRDRPTTGNTLKSGLCSALTTYFFGADLKGKLTIKNFLEFQRKLQHDVLKLEFERHDPVDGRITERQFGGMLLAYSGVQSKKLTAMQRQLKKHFKEGKGLTFQEVENFFTFLKNINDVDTALSFYHMAGASLDKVTMQQVARTVAKVELSDHVCDVVFALFDCDGNGELSNKEFVSIMKQRLMRGLEKPKDMGFTRLMQAMWKCAQETAWDFALPKQ mcu_human Calcium uniporter protein, mitochondrial MCU C10orf42 CCDC109A Coiled-coil domain-containing protein 109A protein protein human Homo sapiens Human MAAAAGRSLLLLLSSRGGGGGGAGGCGALTAGCFPGLGVSRHRQQQHHRTVHQRIASWQNLGAVYCSTVVPSDDVTVVYQNGLPVISVRLPSRRERCQFTLKPISDSVGVFLRQLQEEDRGIDRVAIYSPDGVRVAASTGIDLLLLDDFKLVINDLTYHVRPPKRDLLSHENAATLNDVKTLVQQLYTTLCIEQHQLNKERELIERLEDLKEQLAPLEKVRIEISRKAEKRTTLVLWGGLAYMATQFGILARLTWWEYSWDIMEPVTYFITYGSAMAMYAYFVMTRQEYVYPEARDRQYLLFFHKGAKKSRFDLEKYNQLKDAIAQAEMDLKRLRDPLQVHLPLRQIGEKD emre_human Essential MCU regulator, mitochondrial SMDT1 C22orf32 EMRE Single-pass membrane protein with aspartate-rich tail 1, mitochondrial protein protein human Homo sapiens Human MASGAARWLVLAPVRSGALRSGPSLRKDGDVSAAWSGSGRSLVPSRSVIVTRSGAILPKPVKMSFGLLRVFSIVIPFLYVGTLISKNFAALLEEHDIFVPEDDDDDD micu1-mcu-4 3723830 N/A 3723833 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3723834 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed p.[Lys126Glu;Arg129Glu] mutation with complex effect mutation with complex effect certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q9BPX6:p.[Lys126Ala;Arg129Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q9BPX6:p.[Tyr114Ala;Lys126Ala;Arg129Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false p.[Lys126Ala;Arg129Ala:Tyr114Ala;Tyr121Ala] mutation with complex effect mutation with complex effect certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Q9BPX6:p.Tyr114Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q9BPX6:p.Lys126Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q9BPX6:p.Arg129Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false N/A 3723835 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed c-terminal id4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Supp. Fig 1C, 1D micu1-micu2-2 3723830 N/A 3723833 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed Q8IYU8:p.Arg352Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false N/A 3723834 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Q9BPX6:p.Cys463Ser mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Supp. Fig 1F The inhibitory effect of the MICU2 Arg352Glu mutant was only examined in a MICU1 Cys463Ser background mcu-micu1-9 3723830 N/A 3723836 antibody detection identification by antibody antibody detection unspecified role unspecified role prey prey over-expressed over expressed level over-expressed N/A 3723833 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3723834 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3723835 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal id4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Supp. Figs 1E, 7A, 7B Interaction observed in low calcium conditions mcu-smdt1-1 3723830 N/A 3723836 antibody detection identification by antibody antibody detection unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3723835 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal id4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Supp. Fig 1E, 7A, 7B Interaction observed in high (2mM) calcium conditions. mcu-smdt1-2 3723830 N/A 3723836 antibody detection identification by antibody antibody detection unspecified role unspecified role prey prey over-expressed over expressed level over-expressed N/A 3723835 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal id4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Supp. Fig 1E, 7B Charge-reversal of MICU1’s KKKKR sequence to QEQEQ (EQ) greatly weakens MICU1’s association with MCU although trace amounts of MCU remain associated with the complex. Interaction observed in high salt conditions, under low salt trace amounts of charged reversed MICU1 and also MICU2 were observed in the complex. mcu-micu2-1 3723830 N/A 3723833 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3723836 antibody detection identification by antibody antibody detection unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3723835 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal id4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Supp. Fig 7A Interaction observed in low calcium conditions. micu1-micu2-3 3723830 N/A 3723833 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3723834 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Supp. Fig 1D mcu-micu1-6 3723831 N/A 3723835 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 4.0 N/A 3723836 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 4.0 N/A 3723834 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 3723833 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 physical association physical association false false false Low Calcium Extended Data Table 1 mcu-micu1-7 3723831 N/A 3723835 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 8.0 N/A 3723836 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 8.0 N/A 3723834 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 N/A 3723833 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 physical association physical association false false false High Calcium (2mM). The structure forms a back-to-back dimer. Extended Data Table 1 mcu-micu1-8 3723832 N/A 3723835 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 4.0 N/A 3723836 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Stoichiometry: 4.0 N/A 3723834 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 3723833 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q8IYU8:p.Lys121Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q8IYU8:p.[Arg107Glu;Arg120_Lys121delinsGluGlu;Asp154Arg] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 physical association physical association false false false Low Calcium. The presence of a MCU Asp123Arg mutation changes the complex from a dimeric to a monomeric assembly Supp. Fig. 1A, 1B, 7C, 7D