UniProt UniProt http://www.uniprot.org Structure and mechanism of the mitochondrial Ca2+ uniporter holocomplex. Structure and mechanism of the mitochondrial Ca2+ uniporter holocomplex. Nature 2020 imex curation Fan M., Zhang J., Tsai CW., Orlando BJ., Rodriguez M., Xu Y., Liao M., Tsai MF., Feng L. 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 weight by comassie molecular weight estimation by coomasie staining weight by comassie Fan M., Zhang J., Tsai CW., Orlando BJ., Rodriguez M., Xu Y., Liao M., Tsai MF., Feng L. Accepted 2020-OCT-01 AT 09:33 BST AT 09:33 BST by MGT Nature 2020 Only protein-protein interactions imex curation imex curation Structure and mechanism of the mitochondrial Ca2+ uniporter holocomplex. Structure and mechanism of the mitochondrial Ca2+ uniporter holocomplex. Nature 2020 imex curation Fan M., Zhang J., Tsai CW., Orlando BJ., Rodriguez M., Xu Y., Liao M., Tsai MF., Feng L. 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-01 AT 09:33 BST AT 09:33 BST by MGT Nature 2020 Only protein-protein interactions imex curation imex curation Structure and mechanism of the mitochondrial Ca2+ uniporter holocomplex. Structure and mechanism of the mitochondrial Ca2+ uniporter holocomplex. Nature 2020 imex curation Fan M., Zhang J., Tsai CW., Orlando BJ., Rodriguez M., Xu Y., Liao M., Tsai MF., Feng L. 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. Accepted 2020-OCT-01 AT 09:33 BST AT 09:33 BST by MGT 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 37F31B7BE2DE80E7 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 4E00824D322AE99D mcu_human Calcium uniporter protein, mitochondrial MCU C10orf42 CCDC109A Coiled-coil domain-containing protein 109A protein protein human Homo sapiens Human MAAAAGRSLLLLLSSRGGGGGGAGGCGALTAGCFPGLGVSRHRQQQHHRTVHQRIASWQNLGAVYCSTVVPSDDVTVVYQNGLPVISVRLPSRRERCQFTLKPISDSVGVFLRQLQEEDRGIDRVAIYSPDGVRVAASTGIDLLLLDDFKLVINDLTYHVRPPKRDLLSHENAATLNDVKTLVQQLYTTLCIEQHQLNKERELIERLEDLKEQLAPLEKVRIEISRKAEKRTTLVLWGGLAYMATQFGILARLTWWEYSWDIMEPVTYFITYGSAMAMYAYFVMTRQEYVYPEARDRQYLLFFHKGAKKSRFDLEKYNQLKDAIAQAEMDLKRLRDPLQVHLPLRQIGEKD 50205D01055D66E4 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 DIP protein Q9BPX6 original sequence version: 90 DIP protein Q9BPX6 original sequence version: 135 D83C3A38F4B28FDF mcu-micu1-6 3 6 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 5 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 c-terminal strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 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 4 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 physical association physical association Low Calcium Extended Data Table 1 mcu-micu1-7 3 6 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 5 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 c-terminal strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 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 4 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 physical association physical association High Calcium (2mM). The structure forms a back-to-back dimer. Extended Data Table 1 mcu-micu1-8 1 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp 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 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp 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 c-terminal strep tag strep tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp 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 4 weight by comassie molecular weight estimation by coomasie staining weight by comassie structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp 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 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 K A 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 R E certain certain sequence position certain certain certain sequence position certain RK EE certain certain sequence position certain certain certain sequence position certain D R Wed Sep 16 10:52:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'Q8IYU8:p.[Arg107Glu;Arg120Glu;Lys121Glu:Asp154Arg]' physical association physical association 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 micu1-mcu-4 2 4 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 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 K E certain certain sequence position certain certain certain sequence position certain R E 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 K A certain certain sequence position certain certain certain sequence position certain R A Wed Sep 16 10:43:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys126Ala;Arg129Ala]' 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 K A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain Y A Thu Sep 17 11:15:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys126Ala;Arg129Ala:Tyr114Ala]' p.[Lys126Ala;Arg129Ala:Tyr114Ala;Tyr121Ala] mutation with complex effect mutation with complex effect certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain Y A certain certain sequence position certain certain certain sequence position certain Y A 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 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 Y A Wed Sep 16 11:03:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr114Ala' 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 K E Wed Sep 16 11:02:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys126Glu' 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 R E Fri Sep 18 23:17:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg129Glu' 6 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 association association Supp. Fig 1C, 1D micu1-micu2-2 2 4 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 R E Wed Sep 16 10:16:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg352Glu' 7 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 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 C S Wed Sep 16 10:10:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys463Ser' physical association physical association Supp. Fig 1F The inhibitory effect of the MICU2 Arg352Glu mutant was only examined in a MICU1 Cys463Ser background mcu-micu1-9 2 5 antibody detection identification by antibody antibody detection western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 4 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 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 6 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 association association Supp. Figs 1E, 7A, 7B Interaction observed in low calcium conditions mcu-smdt1-1 2 5 antibody detection identification by antibody antibody detection western blot western blot Immuno blot Immuno blot 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 6 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 physical association physical association Supp. Fig 1E, 7A, 7B Interaction observed in high (2mM) calcium conditions. mcu-smdt1-2 2 5 antibody detection identification by antibody antibody detection western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 6 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 physical association physical association 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 2 4 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 5 antibody detection identification by antibody antibody detection western blot western blot Immuno blot Immuno blot 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 6 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 association association Supp. Fig 7A Interaction observed in low calcium conditions. micu1-micu2-3 2 4 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Supp. Fig 1D