IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. Cell (0092-8674) 2009 imex curation Wang X., Schwarz TL. Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wang X., Schwarz TL. Tagged proteins were identified by anti-tag antibodies Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu Accepted 2009-APR-4 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 9 imex curation The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. Cell (0092-8674) 2009 imex curation Wang X., Schwarz TL. Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Wang X., Schwarz TL. Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu Accepted 2009-APR-4 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 9 imex curation The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. Cell (0092-8674) 2009 imex curation Wang X., Schwarz TL. Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. solution sedimentati cosedimentation in solution solution sedimentati western blot western blot Immuno blot Immuno blot Wang X., Schwarz TL. Cell lysates were incubated with 20 mM Taxol and 0.1 mg/ml polymerized microtubules for 30 min at room temperature without ATP. The microtubule pellet and bound proteins were collected by centrifugation at 21,000 g for 30 min through a 10% sucrose cushion. Tagged proteins were identified by anti-tag antibodies Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu Accepted 2009-APR-4 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 9 imex curation The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. The mechanism of Ca2+ -dependent regulation of kinesin-mediated mitochondrial motility. Cell (0092-8674) 2009 imex curation Wang X., Schwarz TL. Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 9 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wang X., Schwarz TL. Source of KIF5C taken from PMID: 16835241 as given in the text thomas.schwarz@childrens.harvard.edu Accepted 2009-APR-4 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 9 imex curation miro1_human Mitochondrial Rho GTPase 1 Ras homolog gene family member T1 Rac-GTP-binding protein-like protein RHOT1 ARHT1 protein protein human Homo sapiens Human MKKDVRILLVGEPRVGKTSLIMSLVSEEFPEEVPPRAEEITIPADVTPERVPTHIVDYSEAEQSDEQLHQEISQANVICIVYAVNNKHSIDKVTSRWIPLINERTDKDSRLPLILVGNKSDLVEYSSMETILPIMNQYTEIETCVECSAKNLKNISELFYYAQKAVLHPTGPLYCPEEKEMKPACIKALTRIFKISDQDNDGTLNDAELNFFQRICFNTPLAPQALEDVKNVVRKHISDGVADSGLTLKGFLFLHTLFIQRGRHETTWTVLRRFGYDDDLDLTPEYLFPLLKIPPDCTTELNHHAYLFLQSTFDKHDLDRDCALSPDELKDLFKVFPYIPWGPDVNNTVCTNERGWITYQGFLSQWTLTTYLDVQRCLEYLGYLGYSILTEQESQASAVTVTRDKKIDLQKKQTQRNVFRCNVIGVKNCGKSGVLQALLGRNLMRQKKIREDHKSYYAINTVYVYGQEKYLLLHDISESEFLTEAEIICDVVCLVYDVSNPKSFEYCARIFKQHFMDSRIPCLIVAAKSDLHEVKQEYSISPTDFCRKHKMPPPQAFTCNTADAPSKDIFVKLTTMAMYPHVTQADLKSSTFWLRASFGATVFAVLGFAMYKALLKQR DIP protein Q8IXI2 original sequence version: 111 FE47AC3A4AF6F6C2 trak1_human Trafficking kinesin-binding protein 1 Protein Milton TRAK1 KIAA1042 OIP106 106 kDa O-GlcNAc transferase-interacting protein protein protein human Homo sapiens Human MALVFQFGQPVRAQPLPGLCHGKLIRTNACDVCNSTDLPEVEIISLLEEQLPHYKLRADTIYGYDHDDWLHTPLISPDANIDLTTEQIEETLKYFLLCAERVGQMTKTYNDIDAVTRLLEEKERDLELAARIGQSLLKKNKTLTERNELLEEQVEHIREEVSQLRHELSMKDELLQFYTSAAEESEPESVCSTPLKRNESSSSVQNYFHLDSLQKKLKDLEEENVVLRSEASQLKTETITYEEKEQQLVNDCVKELRDANVQIASISEELAKKTEDAARQQEEITHLLSQIVDLQKKAKACAVENEELVQHLGAAKDAQRQLTAELRELEDKYAECMEMLHEAQEELKNLRNKTMPNTTSRRYHSLGLFPMDSLAAEIEGTMRKELQLEEAESPDITHQKRVFETVRNINQVVKQRSLTPSPMNIPGSNQSSAMNSLLSSCVSTPRSSFYGSDIGNVVLDNKTNSIILETEAADLGNDERSKKPGTPGTPGSHDLETALRRLSLRRENYLSERRFFEEEQERKLQELAEKGELRSGSLTPTESIMSLGTHSRFSEFTGFSGMSFSSRSYLPEKLQIVKPLEGSATLHHWQQLAQPHLGGILDPRPGVVTKGFRTLDVDLDEVYCLNDFEEDDTGDHISLPRLATSTPVQHPETSAHHPGKCMSQTNSTFTFTTCRILHPSDELTRVTPSLNSAPTPACGSTSHLKSTPVATPCTPRRLSLAESFTNTRESTTTMSTSLGLVWLLKERGISAAVYDPQSWDRAGRGSLLHSYTPKMAVIPSTPPNSPMQTPTSSPPSFEFKCTSPPYDNFLASKPASSILREVREKNVRSSESQTDVSVSNLNLVDKVRRFGVAKVVNSGRAHVPTLTEEQGPLLCGPPGPAPALVPRGLVPEGLPLRCPTVTSAIGGLQLNSGIRRNRSFPTMVGSSMQMKAPVTLTSGILMGAKLSKQTSLR E834BE07CF41D9A4 kif5c_rat Kinesin heavy chain isoform 5C Kinesin heavy chain neuron-specific 2 Kif5c Nkhc2 protein protein rat Rattus norvegicus (Rat) Rat MADPAECSIKVMCRFRPLNEAEILRGDKFIPKFKGEETVVIGQGKPYVFDRVLPPNTTQEQVYNACAKQIVKDVLEGYNGTIFAYGQTSSGKTHTMEGKLHDPQLMGIIPRIAHDIFDHIYSMDENLEFHIKVSYFEIYLDKIRDLLDVSKTNLAVHEDKNRVPYVKGCTERFVSSPEEVMDVIDEGKANRHVAVTNMNEHSSRSHSIFLINIKQENVETEKKLSGKLYLVDLAGSEKV C96C5FFE4F8E1D7E kinh_rat Kinesin-1 heavy chain Kif5b Ubiquitous kinesin heavy chain Conventional kinesin heavy chain Khc protein protein rat Rattus norvegicus (Rat) Rat MADPAECNIKVMCRFRPLNESEVNRGDKYVAKFQGEDTVMIASKPYAFDRVFQSSTSQEQVYNDCAKKIVKDVLEGYNGTIFAYGQTSSGKTHTMEGKLHDPEGMGIIPRIVQDIFNYIYSMDENLEFHIKVSYFEIYLDKIRDLLDVSKTNLSVHEDKNRVPYVKGCTERFVCSPDEVMDTIDEGKSNRHVAVTNMNEHSSRSHSIFLINVKQENTQTEQKLSGKLYLVDLAGSEKVSKTGAEGAVLDEAKNINKSLSALGNVISALAEGSTYVPYRDSKMTRILQDSLGGNCRTTIVICCSPSSYNESETKSTLLFGQRAKTIKNTVCVNVELTAEQWKKKYEKEKEKNKTLRNTIQWLENELNRWRNGETVPIDEQFDKEKANLEAFTADKDVAITNDKPAAAIGMAGSFTDAERRKCEEEIAKLYKQLDDKDEEINQQSQLVEKLKTQMLDQEELLASTRRDQDNMQAELNRLQAENDASKEEVKEVLQALEELAVNYDQKSQEVEDKTKEYELLSDELNQKSATLASIDAELQKLKEMTNHQKKRAAEMMASLLKDLAEIGIAVGNNDVKQPEGTGMIDEEFTVARLYISKMKSEVKTMVKRCKQLESTQTESNKKMEENEKELAACQLRISQHEAKIKSLTEYLQNVEQKKRQLEESVDSLGEELVQLRAQEKVHEMEKEHLNKVQTANEVKQAVEQQIQSHRETHQKQISSLRDEVEAKEKLITDLQDQNQKMVLEQERLRVEHERLKAVDQEKSRKLHELTVMQDRREQARQDLKGLEETVAKELQTLHNLRKLFVQDLATRVKKSAEVDSDDTGGSAAQKQKISFLENNLEQLTKVHKQLVRDNADLRCELPKLEKRLRATAERVKALESALKEAKENASRDRKRYQQEVDRIKEAVRSKNMARRGHSAQIAKPIRPGQHPAASPTHPGAVRGGGSFVQNNQPVGLRGGGGKQA DIP protein Q2PQA9 original sequence version: 30 0A460B2CBA47EC30 kif5c_human Kinesin heavy chain isoform 5C Kinesin heavy chain neuron-specific 2 KIF5C KIAA0531 NKHC2 protein protein human Homo sapiens Human MADPAECSIKVMCRFRPLNEAEILRGDKFIPKFKGDETVVIGQGKPYVFDRVLPPNTTQEQVYNACAKQIVKDVLEGYNGTIFAYGQTSSGKTHTMEGKLHDPQLMGIIPRIAHDIFDHIYSMDENLEFHIKVSYFEIYLDKIRDLLDVSKTNLAVHEDKNRVPYVKGCTERFVSSPEEVMDVIDEGKANRHVAVTNMNEHSSRSHSIFLINIKQENVETEKKLSGKLYLVDLAGSEKVSKTGAEGAVLDEAKNINKSLSALGNVISALAEGTKTHVPYRDSKMTRILQDSLGGNCRTTIVICCSPSVFNEAETKSTLMFGQRAKTIKNTVSVNLELTAEEWKKKYEKEKEKNKTLKNVIQHLEMELNRWRNGEAVPEDEQISAKDQKNLEPCDNTPIIDNIAPVVAGISTEEKEKYDEEISSLYRQLDDKDDEINQQSQLAEKLKQQMLDQDELLASTRRDYEKIQEELTRLQIENEAAKDEVKEVLQALEELAVNYDQKSQEVEDKTRANEQLTDELAQKTTTLTTTQRELSQLQELSNHQKKRATEILNLLLKDLGEIGGIIGTNDVKTLADVNGVIEEEFTMARLYISKMKSEVKSLVNRSKQLESAQMDSNRKMNASERELAACQLLISQHEAKIKSLTDYMQNMEQKRRQLEESQDSLSEELAKLRAQEKMHEVSFQDKEKEHLTRLQDAEEMKKALEQQMESHREAHQKQLSRLRDEIEEKQKIIDEIRDLNQKLQLEQEKLSSDYNKLKIEDQEREMKLEKLLLLNDKREQAREDLKGLEETVSRELQTLHNLRKLFVQDLTTRVKKSVELDNDDGGGSAAQKQKISFLENNLEQLTKVHKQLVRDNADLRCELPKLEKRLRATAERVKALESALKEAKENAMRDRKRYQQEVDRIKEAVRAKNMARRAHSAQIAKPIRPGHYPASSPTAVHAIRGGGGSSSNSTHYQK A9F25BB1C994322A miro_drome Mitochondrial Rho GTPase CG5410 Miro protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MGQYTASQRKNVRILLVGDAGVGKTSLILSLVSEEYPEEVPPRAEEITIPANVTPEQVPTSIVDFSAVEQSEDALAAEINKAHVVCIVYAVDDDDTLDRITSHWLPLVRAKCNPSLDGEGDAEAEAEGDTQREPIRKPIVLVGNKIDLIEYSTMDSVLAIMEDYPEIESCVECSAKSLHNISEMFYYAQKAVLHPTSPLYMMEEQELTSACKKSLVRIFKICDIDGDNLLNDYELNLFQRRCFNTPLQPQILDEVKAVIQKNVPDGIYNDAVTLKGFLFLHCLFIQRGRNETTWAVLRRFGYNDQLEMCQEYLRPPLKIPPGSSTELSHRGQQFLIAVFERYDRDGDGALSPEEHKMLFSTCPAAPWSYSTDIRKSCPINETTGWVTLHGWLCRWTLMTLIDVVKTMEYLAYLGFNVHENDSQLAAIHVTRERRIDLAKRQSSRSVYKCHVIGPKGSGKTGMCRGFLVEDMHKLIGKEFKTNVVNCINSVQVYGQEKHLILRDIDVRHALDPLQPQEVNCDVACLVYDSSNPRSFEYVARIYIKYYAESKIPVMIVGTKCDMDERRQDYLMQPSEFCDKYKLLPPHLFSLKTNKKELYTKLATMAAFPHLRQFGLMTEDPKLWLKAGLGVAAATMLGFIVLKTISAAGAHTR 5F2F9623EF6CCB19 milt_drome Trafficking kinesin-binding protein milt milt Protein milton CG43227 protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MTHVNNGEVMEKEMEPTGERERDRETAAGSGVHHRFLASASERDANSGNALEAAATKTKTGTTITNLEDLAFEACQNWSDLHQDFFITDDELEYEDELSLGSSIGGNIATTTTADAAVEGLITGEHQNEQLLMEVLCGNRVSQMTRAYDDIEAVTRLLEEKEKDLELTVQIGKELLTQNNALEARVADLETDLKASNDDRAQLVHELHKKNELISVLTNDADDGTDTDTPTMSKSITLDLLQRKVNSLLDENKSLKCEATQLAHQTDEVEEHERQLMADISAQLNDANSQYDNLSLELERQREENRLQHEQIVNLTARLAEAEMRLHQLTQDNDEHLSLLHVTKENQNALALELVEFKQRYEEVLALLHSAQDQLKQQRKRSQPQARSSFLGGLGTSGAGMGGSLFPPDSLHCELMESSLYSENSLDSGISGDSQRSADRISRMMMHMPSGGMSSSTMGGSVYAGAGNVPPYKRVFDTVRCAGKSGNYMDSGNVSMTQLGAMSMSSSSGPRMASMAYPAGSYYRGGSNQSLGVKTLSSESLNSQSDDGYPAQPSGVPGAPGAKELEAALKRLTPAEVLARRAMLSYAPAGTYNYDEPMGHGTGNVRNSDLPLGVRTPDSIMSTGSSGMSGSTNHMSASMTHQWRLPEKLQIIKPMEGSQTLHHWSRLATPTLSGLLDERPGVTIRGGRGLDDLGMQIYTLSDVEEDVSDDLPGKQFEAPGCTFTYTNSMVMHPDDGFVNDLSFLSQSQMSSRMASTSTSRQPSCPATPRAGLSRKNSCSTFSVNLGLAGMLNERGIKAVTPSALNTPAGPNFSPTVTPCNSPEGSPPRAQSPEPLFGLLSCGADLIRRKIVGDQHQQQQQKQRSSLSKQQQQKIMLSHLERRALRSLNLIEKVESIGLENIISAQRGLGSGIANRSSSPLSSGSLQSLHTSSNSIVDDIHFDRAQIKGVLHRGLKSPTPATASTSAAAAASGPGISTSSSTSAYNSDDSDDQGLVMKIKPSKSATPTTTGAAQSQPSSATTSNGTASETRLKQMQRQKSRRQLKNGMANQRPDLGTISGAGGGGRVRPDLGKVADSGSSSKLSTKRSEAKPAEEEEATPQTITQAFVGSVSSLLFGRKGGWL 7BBE5E474C8112BC rhot1-trak1-4 4 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey association association Kinesin heavy chains were also identified in the immunoprecipitate but could not be ascribed to a specific gene product The endogenous motor/adaptor complex from HEK293T cells was immunoprecipitated with anti-hMiro1 and probed with anti-KHC, anti-hMiro1, and anti-OIP106 (one of two human homologs of milton). Fig. 1B rhot1-trak1-3 1 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey association association The co-immunoprecipitation of TRAK1 (MILTON) and KHC from HEK cells requires the presence of RHOT1 (MIRO). The co-immunoprecipitation of Miro-Myc and KHC-ECFP from HEK cells requires milton. Figs. 1D, 4G, 6A The interaction is not calcium dependent rhot1-trak1-5 3 5 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IXI2:p.[Glu208Lys;Glu328Lys] EF-hand-disruption mutant mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain E K certain certain sequence position certain certain certain sequence position certain E K Fri Jul 22 11:04:36 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu328lys,glu208lys' Wed May 02 15:01:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu208lys,glu328lys' Tue Jun 30 16:51:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu208Lys;Glu328Lys]' 11 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization When KHC was present without cotransfected milton and Miro, it was exclusively found in the microtubule pellet, whether or not Ca2+ was present. When KHC was coexpressed with milton and Miro, however, Ca2+ caused a substantial shift of KHC, together with the adaptor proteins, to the soluble fraction. When KHC was coexpressed with milton and Miro, however, Ca2+ caused a substantial shift of KHC, together with the adaptor proteins, to the soluble fraction. Fig. 5 rhot1-kif5c-2 1 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IXI2:p.[Glu208Lys;Glu328Lys] EF-hand-disruption mutant mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain E K certain certain sequence position certain certain certain sequence position certain E K Wed May 02 20:12:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu208lys,glu328lys' Tue Jun 30 16:09:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu208Lys;Glu328Lys]' 7 unspecified role unspecified role bait bait region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The co-immunoprecipitation of RHOT1 (MIRO) and KHC from HEK cells requires the presence of calcium. We investigated the possibility that Ca2+ induced a direct interaction of Miro and KHC, and found that Miro and KHC would indeed co-precipitate in elevated Ca2+ Figs. 6A rhot1-kif5b-1 1 8 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role prey prey region transmembrane domain deleted sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range certain certain sequence position certain certain certain sequence position certain range range region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IMX7:p.[Glu234Lys;Asp345Lys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D K certain certain sequence position certain certain certain sequence position certain E K Fri Jul 22 11:10:12 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu234lys_glu354lys' Wed May 02 15:15:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu234lys,asp345lys' Tue Jun 30 18:02:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu234Lys;Asp345Lys]' physical association physical association The co-immunoprecipitation of Miro and Kif5b from HEK cells requires the presence of calcium. Immunoprecipitations with anti-Myc from cells expressing Myc-tagged full length (FL, left two panels) or truncated KHC 1-682 (682, right two panels), and T7-tagged DMiroΔTM, but not milton. Figs. 6D Exact coordinates of the transmembrane domain of Miro were not given in the paper and have been inferred from the UniProtKB entry. rhot1-kif5c-3 1 5 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IXI2:p.[Glu208Lys;Glu328Lys] EF-hand-disruption mutant mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain E K certain certain sequence position certain certain certain sequence position certain E K Fri Jul 22 10:50:06 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu328lys,glu208lys' Wed May 02 19:16:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu208lys,glu328lys' Tue Jun 30 17:59:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu208Lys;Glu328Lys]' 7 unspecified role unspecified role bait bait region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The co-immunoprecipitation of RHOT1 (MIRO) and KHC from HEK cells requires the presence of calcium. We investigated the possibility that Ca2+ induced a direct interaction of Miro and KHC, and found that Miro and KHC would indeed co-precipitate in elevated Ca2+ Figs. 6A rhot1-trak1-9 3 5 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IXI2:p.[Glu208Lys;Glu328Lys] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain E K certain certain sequence position certain certain certain sequence position certain E K Wed May 02 17:49:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu208lys,glu328lys' Tue Jun 30 16:17:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu208Lys;Glu328Lys]' 9 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association The co-immunoprecipitation of RHOT1 (MIRO) and KHC from HEK cells requires the presence of calcium. Indeed, when a construct encoding only the N-terminal motor domain of KHC was coexpressed with Miro they exhibited Ca2+-dependent coprecipitation (Figures 6G and 6H). Figs. 6I, 6J, 6K rhot1-kifc-1 1 5 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The co-immunoprecipitation of RHOT1 (MIRO) and KHC from HEK cells requires the presence of calcium. Ca2+ dependence of the binding of Miro-Myc to KIF5C(1-335)-YFP, by precipitation from lysates of transfected HEK cells with anti-Myc and probing with anti-GFP (KIF5C(1-335)-YFP). Figs. 6G, 6L rhot1-if5c-2 2 5 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role neutral component neutral component region mCit tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Co-localisation requires the presence of a drsophila Milton orthologue KHC-mCit is diffusely cytoplasmic when only Miro has been expressed but colocalizes in puncta with MitoTracker and Miro when milton is also expressed. Supp. Fig. 2