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/ http://www.ebi.ac.uk/ wang-2008f-4 Myosin Vb mobilizes recycling endosomes and AMPA receptors for postsynaptic plasticity. rat-hippocampus Rattus norvegicus hippocampal neurons hippocampus Hippocampus [Brain hippocampus] fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Wang Z., Edwards JG., Riley N., Provance DW., Karcher R., Li XD., Davison IG., Ikebe M., Mercer JA., Kauer JA., Ehlers MD. Synapse - Interactions of proteins with an established role in the presynapse. ehlers@neuro.duke.edu Cell (0092-8674) 2008 Only protein-protein interactions 7 imex curation wang-2008f-3 Myosin Vb mobilizes recycling endosomes and AMPA receptors for postsynaptic plasticity. rat-brain Rattus norvegicus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wang Z., Edwards JG., Riley N., Provance DW., Karcher R., Li XD., Davison IG., Ikebe M., Mercer JA., Kauer JA., Ehlers MD. Synapse - Interactions of proteins with an established role in the presynapse. ehlers@neuro.duke.edu Anti-Rab11 identified as anti-Rab11a in PMID: 12376546 Cell (0092-8674) 2008 Only protein-protein interactions 7 imex curation wang-2008f-1 Myosin Vb mobilizes recycling endosomes and AMPA receptors for postsynaptic plasticity. rat-hippocampus Rattus norvegicus hippocampal neurons hippocampus Hippocampus [Brain hippocampus] fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Wang Z., Edwards JG., Riley N., Provance DW., Karcher R., Li XD., Davison IG., Ikebe M., Mercer JA., Kauer JA., Ehlers MD. Synapse - Interactions of proteins with an established role in the presynapse. ehlers@neuro.duke.edu Cell (0092-8674) 2008 Only protein-protein interactions 7 imex curation wang-2008f-2 Myosin Vb mobilizes recycling endosomes and AMPA receptors for postsynaptic plasticity. in vitro In vitro atpase assay atpase assay predetermined predetermined participant predetermined Wang Z., Edwards JG., Riley N., Provance DW., Karcher R., Li XD., Davison IG., Ikebe M., Mercer JA., Kauer JA., Ehlers MD. Synapse - Interactions of proteins with an established role in the presynapse. ehlers@neuro.duke.edu Cell (0092-8674) 2008 Only protein-protein interactions 7 imex curation wang-2008f-5 Myosin Vb mobilizes recycling endosomes and AMPA receptors for postsynaptic plasticity. 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 anti tag western anti tag western blot anti tag western Wang Z., Edwards JG., Riley N., Provance DW., Karcher R., Li XD., Davison IG., Ikebe M., Mercer JA., Kauer JA., Ehlers MD. Synapse - Interactions of proteins with an established role in the presynapse. Source of RAB11FIP2 taken from PMID: 15173169. ehlers@neuro.duke.edu Cell (0092-8674) 2008 Only protein-protein interactions 7 imex curation myo5b_rat Unconventional myosin-Vb Myosin heavy chain myr 6 Myo5b protein protein rat Rattus norvegicus (Rat) Rat MTYSELYSRYTRVWIPDPDEVWRSAELTKDYKDGDESLQLRLEDDTILDYPIDVQNNQVPFLRNPDILVGENDLTALSHLHEPAVLHNLKVRFLESNHIYTYCGIVLVAINPYEQLPIYGQDVIYAYSGQNMGDMDPHIFAVAEEAYKQMARDEKNQSIIVSGESGAGKTVSAKYAMRYFATVGGSASDTNIEEKVLASSPIMEAIGNAKTTRNDNSSRFGKYIEIGFDKKYHIIGANMRTYLLEKSRVVFQADDERNYHIFYQLCAAASLPEFKELALTCAEDFFYTAHGGNTTIEGVDDAEDFEKTRQALTLLGVRESHQISIFKIIASILHLGSVEIQAERDGDSCSISPQDEHLSNFCRLLGIEHSQMEHWLCHRKLVTTSETYVKTMSLQQVVNARNALAKHIYAQLFSWIVEHINKALQTSLKQHSFIGVLDIYGFETFEINSFEQFCINYANEKLQQQFNSHVFKLEQEEYMKEQIPWTLIDFYDNQPCIDLIEAKLGILDLLDEECKVPKGTDQNWAQKLYERHSNSQHFQKPRMSNTAFIVIHFADKVEYLSDGFLEKNRDTVYEEQINILKASKFPLVADLFRDDEDSVPATNTAKSRSSSKINVRSSRPLMKAPNKEHKKSVGYQFRTSLNLLMETLNATTPHYVRCIKPNDEKLPFHFDPKRAVQQLRACGVLETIRISAAGYPSRWTYHDFFNRYRVLMKKRELANTTDKKNICKSVLESLIKDPDKFQFGRTKIFFRAGQVAYLEKLRADKFREATIMIQKTVRGWLQRVKYRRLRAATLTLQRFCRGYLARRLTEHLRRTRAAIVFQKQYRMLKARRAYCRVRRAAVIIQSYTRGHVCTQKLPPVLTEHKATIIQKYARGWMARRHFQRQRDAAIVIQCAFRRLKARQALKALKIEARSAEHLKRLNVGMENKVVQLQRKIDDQNKEFKTLSEQLSAVTSTHAMEVEKLKKELARYQQNQEADPSLQLQEEVQSLRTELQKAHSERRVLEDAHNRENGELRKRVADLEHENALLKDEKEHLNHQILRQSKAESSQSSVEENLLIKKELEEERSRYQNLVKEYSQLEQRYENLRDEQQTPGHRKNPSNQSSLESDSNYPSISTSEIGDTEDALQQVEEIGIEKAAMDMTVFLKLQKRVRELEQERKKLQVQLEKEQQDSKKVQVEQQNNGLDVDQDADIAYNSLKRQELESENKKLKNDLNERWKAVADQAMQDNSTHSSPDSYSLLLNQLKLANEELEVRKEEVLILRTQIMNADQRRLSGKNMEPNINARTSWPNSEKHVDQEDAIEAYHGVCQTNSQTEDWGYLNEDGELGLAYQGLKQVARLLEAQLQAQNLKHEEEVEHLKAQVEAMKEEMDKQQQTFCQTLLLSPEAQVEFGVQQEISRLTNENLDFKELVEKLEKNEKKLKKQLKIYMKKVQDLEAAQALAQSDRRHHELTRQVTVQRKEKDFQGMLEYHKEDEALLIRNLVTDLKPQMLSGTVPCLPAYILYMCIRHADYTNDDLKVHSLLSSTINGIKKVLKKHNEDFEMTSFWLSNTCRLLHCLKQYSGDEGFMTQNTAKQNEHCLKNFDLTEYRQVLSDLSIQIYQQLIKIAEGLLQPMIVSAMLENESIQGLSGVRPTGYRKRSSSMVDGENSYCLEAIIRQMNFFHTVLCDQGLDPEIILQVFKQLFYMINAVTLNNLLLRKDACSWSTGMQLRYNISQLEEWLRGKNLQQSGAVQTMEPLIQAAQLLQLKKKTQEDAEAICSLCTSLSTQQIVKILNLYTPLNGFEERVTVSFIRTIQAQLQERSDPQQLLLDSKHMFPVLFPFNPSALTMDSIHIPACLNLEFLNEV tfr1_rat Transferrin receptor protein 1 Tfrc Trfr protein protein rat Rattus norvegicus (Rat) Rat MMDQARSAFSNLFGGEPLSYTRFSLARQVDGDNSHVEMKLAADEEENADSNMKASVRKPKRFNGRLCFATIAVVIFFLIGFMIGYLGYCKRVEQKEECVRLAEAEEADKSENDETEYVPKSSRLFWADLKTLLSEKLNSIEFTDIIKQLSQNTYTPREAGSQKDENLAYYIENLFHDFKFSKVWRDEHYVKIQVKNSVSQNLVTINSGSNIDPVEAPEGYVAFSKAGEVTGKLVHANFGTKKDFEELNYSVNGSLVIVRAGKITFAEKVANAQSFNAIGVLIYMDRNTFPVVEADLQFFGHAHLGTGDPYTPGFPSFNHTQFPPSQSSGLPSIPVQTISRAAAEKLFKNMEGNCPPSWNIDSSCKLELSQNQNVKLTVNNVLKETRILNIFGVIKGYEEPDRYIVVGAQRDAWGPGVAKSSVGTGLLLKLAQVFSDMISKDGFRPSRSIIFASWTAGDYGAVGATEWLEGYLSSLHLKAFTYINLDKVVLGTSNFKVSASPLLYTLMGKIMQDVKHPIDGKYLYRDSNWISKIEELSLDNAAFPFLAYSGIPAVSFCFCEDEDYPYLGTKLDTYEILIQKVPQLNQMVRTAAEVAGQFIIKLTHDIELTLDYEMYNSKLLSFMKDLNQFKADIKDMGLSLQWLYSARGDYFRATSRLTTDFHNAEKTNRFVMREINDRIMKVEYHFLSPYVSPRESPFRHIFWGSGSHTLSALVENLRLRQKNITAFNETLFRNQLALATWTIQGVANALSGDIWNIDNEF d3zd48_rat Rab11fip2 Rab11fip2_predicted rCG_57662 protein protein rat Rattus norvegicus (Rat) Rat MMLSEQAQKWFPTHVQVTVLQAKDLKPKGKSGTNDTYTIIQLGKEKYSTSVAEKTLEPVWKEEASFELPGLLMQGNPEKYILFLIVMHRSLVGLDKFLGQVAINLNDIFEDKQRRKTEWFRLESKQGKRIKNRGEIKVNIQFMRNNMTASMFDLSMKDKTRSPFAKLKDKMKGRKSDGVFSDTSSAIVPSTHMPDANPEFSSGEMQMKSKPKKPFLLGPQRLSSAHSMSDLTGSHLSSEKLKSSTVGPTHLLSHQINSFGVVPESGSLKSPHRRTLSFDTSKLNQPGSIVDEGEHCFGRQSDPFTNVTASLPQKFATLPRKKNPFEESSEPWDSSTNLFSKPIEVRKESKREKREKVSLFERVTGKRDSRRSDKLNNGGSDSPCDLKSPSAFNENRQDYFEYESTNPFTAKFRASTIMPSSSFHMNPTSSDDLRKIPDSNPFDATAGYRSLTYEEVLQELVKHKELLRRKDTHIRELEDYIDNLLVRVMEETPSILRVPYEPSRKAGKFTNS rb11a_rat Ras-related protein Rab-11A Rab11a Rab11 24KG protein protein rat Rattus norvegicus (Rat) Rat MGTRDDEYDYLFKVVLIGDSGVGKSNLLSRFTRNEFNLESKSTIGVEFATRSIQVDGKTIKAQIWDTAGQERYRAITSAYYRGAVGALLVYDIAKHLTYENVERWLKELRDHADSNIVIMLVGNKSDLRHLRAVPTDEARAFAEKNGLSFIETSALDSTNVEAAFQTILTEIYRIVSQKQMSDRRENDMSPSNNVVPIHVPPTTENKPKVQCCQNI dlg4_rat Disks large homolog 4 Dlg4 Dlgh4 Postsynaptic density protein 95 Synapse-associated protein 90 Psd95 protein protein rat Rattus norvegicus (Rat) Rat MDCLCIVTTKKYRYQDEDTPPLEHSPAHLPNQANSPPVIVNTDTLEAPGYELQVNGTEGEMEYEEITLERGNSGLGFSIAGGTDNPHIGDDPSIFITKIIPGGAAAQDGRLRVNDSILFVNEVDVREVTHSAAVEALKEAGSIVRLYVMRRKPPAEKVMEIKLIKGPKGLGFSIAGGVGNQHIPGDNSIYVTKIIEGGAAHKDGRLQIGDKILAVNSVGLEDVMHEDAVAALKNTYDVVYLKVAKPSNAYLSDSYAPPDITTSYSQHLDNEISHSSYLGTDYPTAMTPTSPRRYSPVAKDLLGEEDIPREPRRIVIHRGSTGLGFNIVGGEDGEGIFISFILAGGPADLSGELRKGDQILSVNGVDLRNASHEQAAIALKNAGQTVTIIAQYKPEEYSRFEAKIHDLREQLMNSSLGSGTASLRSNPKRGFYIRALFDYDKTKDCGFLSQALSFRFGDVLHVIDAGDEEWWQARRVHSDSETDDIGFIPSKRRVERREWSRLKAKDWGSSSGSQGREDSVLSYETVTQMEVHYARPIIILGPTKDRANDDLLSEFPDKFGSCVPHTTRPKREYEIDGRDYHFVSSREKMEKDIQAHKFIEAGQYNSHLYGTSVQSVREVAEQGKHCILDVSANAVRRLQAAHLHPIAIFIRPRSLENVLEINKRITEEQARKAFDRATKLEQEFTECFSAIVEGDSFEEIYHKVKRVIEDLSGPYIWVPARERL atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) rfip2_human Rab11 family-interacting protein 2 NRip11 RAB11FIP2 KIAA0941 protein protein human Homo sapiens Human MMLSEQAQKWFPTHVQVTVLQAKDLKPKGKSGTNDTYTIIQLGKEKYSTSVAEKTLEPVWKEEASFELPGLLIQGSPEKYILFLIVMHRSLVGLDKFLGQVAINLNDIFEDKQRRKTEWFRLESKQGKRIKNRGEIKVNIQFMRNNMTASMFDLSMKDKTRSPFAKLKDKMKGRKNDGTFSDTSSAIIPSTHMPDANSEFSSGEIQMKSKPKKPFLLGPQRLSSAHSMSDLSGSHMSSEKLKAGTIGQTHLLGHQLDSFGTVPESGSLKSPHRRTLSFDTSKMNQPDSIVDEGELCFGRQNDPFTNVTASLPQKFATLPRKKNPFEESSETWDSSMNLFSKPIEIRKENKREKREKVSLFERVTGKKDSRRSDKLNNGGSDSPCDLKSPNAFSENRQDYFDYESTNPFTAKFRASNIMPSSSFHMSPTSNEDLRKIPDSNPFDATAGYRSLTYEEVLQELVKHKELLRRKDTHIRELEDYIDNLLVRVMEETPSILRVPYEPSRKAGKFSNS tfrc-myo5b-1 Colocalisation of TfRC and Myo5B 777859 n/a 777864 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 777865 unspecified role unspecified role neutral component neutral component region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 1C, 1E A C-terminal domain of MyoVb was shown to be required for the interaction, coordinates not given. rab11fip2-myo5b-2 Colocalisation of Rab11fip2 and Myo5B 777859 n/a 777866 unspecified role unspecified role neutral component neutral component n/a 777864 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Figs. 4A, 4C, 5, Supp. Fig. 3F A C-terminal domain of MyoVb was shown to be required for the interaction, coordinates not given. Myo5B mutants lacking the coiled-coil region (949-1042) and locked in an open conformation demonstrated increased colocalisation Ca2+ Glycine tfrc-rab11fip2-1 Colocalisation of Rab11fip2 and TFRC 777859 n/a 777865 unspecified role unspecified role neutral component neutral component region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 777866 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Supp. Fig. 3C-F myo5b-rab11-1 Interaction between Rab11, Rabllfip2 and Myo5b determined by coimmunoprecipitation 777860 n/a 777866 unspecified role unspecified role prey prey n/a 777867 unspecified role unspecified role prey prey n/a 777864 unspecified role unspecified role bait bait physical association physical association false false false Coimmunoprecipitation (co-IP) revealed that MyoVb forms a complex with Rab11 and Rab11-FIP2 in brain (Figure 3E). Fig. 3E myo5b-rab11-2 Interaction between Rab11, Rab11fip2 and Myo5b determined by coimmunoprecipitation 777860 n/a 777866 unspecified role unspecified role prey prey n/a 777867 unspecified role unspecified role bait bait n/a 777864 unspecified role unspecified role prey prey physical association physical association false false false Coimmunoprecipitation (co-IP) revealed that MyoVb forms a complex with Rab11 and Rab11-FIP2 in brain (Figure 3E). Fig. 3E dlg4-myo5b-1 Colocalisation of Dlg4 and Myo5B 777861 n/a 777864 unspecified role unspecified role neutral component neutral component n/a 777868 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 1B, Supp. Fig. 1C myo5b-atp-1 ATPase activity of Myo5b 777862 n/a 777869 enzyme target enzyme target substrate substrate neutral component neutral component n/a 777864 enzyme enzyme neutral component neutral component n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection atpase reaction atpase reaction false false false Ca2+ The calculated Vmax = 1.17 s1motor-1 and and Kactin = 12.58 uM in the absence of Ca2+ and 12.07 s-1motor-1 and 7.11 uM in the presence of 100 uM Ca2+ Fig. 3A rab11fip2-myo5b-1 Interaction between RAB11FIP2 and Myo5b identified by coimmunoprecipitation 777863 n/a 777864 unspecified role unspecified role prey prey region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n/a 777870 unspecified role unspecified role bait bait region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 3F, 3H Ca2+ A version of Myo5B locked in an open conformation due to deletion of the coiled coil region (949-1042) was able to bind RAB11FIP2 in the absence of Ca2+ suggesting that the Ca2+-dependent extension of MyoVb enables its association with cargo