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/ SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. Cell (0092-8674) 2007 imex curation Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 7 mouse-brain Mus musculus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp Accepted 2007-DEC-21 by DAVET. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 7 imex curation SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. Cell (0092-8674) 2007 imex curation Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 7 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. pull down pull down western blot western blot Immuno blot Immuno blot Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 7 imex curation SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. Cell (0092-8674) 2007 imex curation Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 7 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 Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 7 imex curation SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. SCRAPPER-dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. Cell (0092-8674) 2007 imex curation Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 7 mouse-brain-hippocampus Mus musculus brain_hippocampus brain hippocampus fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Yao I., Takagi H., Ageta H., Kahyo T., Sato S., Hatanaka K., Fukuda Y., Chiba T., Morone N., Yuasa S., Inokuchi K., Ohtsuka T., Macgregor GR., Tanaka K., Setou M. Synapse - Interactions of proteins with an established role in the presynapse. Authors have identified a new isoform of Fbxll20 - this is currently being added to the existing UniProt entry after which the entry will be remapped to a more granular protein description. setou@nips.ac.jp Accepted 2007-DEC-21 by DAVET. 22-Jan-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 7 imex curation cul1_mouse Cullin-1 Cul1 protein protein mouse Mus musculus Mouse MSSNRSQNPHGLKQIGLDQIWDDLRAGIQQVYTRQSMAKSRYMELYTHVYNYCTSVHQSNQARGAGVPPSKSKKGQTPGGAQFVGLELYKRLKEFLKNYLTNLLKDGEDLMDESVLKFYTQQWEDYRFSSKVLNGICAYLNRHWVRRECDEGRKGIYEIYSLALVTWRDCLFRPLNKQVTNAVLKLIEKERNGETINTRLISGVVQSYVELGLNEDDAFAKGPTLTVYKESFESQFLADTERFYTRESTEFLQQNPVTEYMKKAEARLLEEQRRVQVYLHESTQDELARKCEQVLIEKHLEIFHTEFQNLLDADKNEDLGRMYNLVSRIQDGLGELKKLLETHIHNQGLAAIEKCGEAALNDPKMYVQTVLDVHKKYNALVMSAFNNDAGFVAALDKACGRFINNNAVTKMAQSSSKSPELLARYCDSLLKKSSKNPEEAELEDTLNQVMVVFKYIEDKDVFQKFYAKMLAKRLVHQNSASDDAEASMISKLKQACGFEYTSKLQRMFQDIGVSKDLNEQFKKHLTNSEPLDLDFSIQVLSSGSWPFQQSCTFALPSELERSYQRFTAFYASRHSGRKLTWLYQLSKGELVTNCFKNRYTLQASTFQMAILLQYNTEDAYTVQQLTDSTQIKMDILAQVLQILLKSKLLVLEDENANVDEVELKPDTLIKLYLGYKNKKLRVNINVPMKTEQKQEQETTHKNIEEDRKLLIQAAIVRIMKMRKVLKHQQLLGEVLTQLSSRFKPRVPVIKKCIDILIEKEYLERVDGEKDTYSYLA 3E1F8B63C6E498F4 3E1F8B63C6E498F4 fxl20_mouse F-box/LRR-repeat protein 20 Fbxl20 Fbl2 F-box and leucine-rich repeat protein 20 F-box/LRR-repeat protein 2-like protein protein mouse Mus musculus Mouse MRRDVNGVTKSRFEMFSNSDEAVINKKLPKELLLRIFSFLDVVTLCRCAQVSRAWNVLALDGSNWQRIDLFDFQRDIEGRVVENISKRCGGFLRKLSLRGCLGVGDNALRTFAQNCRNIEVLSLNGCTKTTDATCTSLSKFCSKLRHLDLASCTSITNMSLKALSEGCPLLEQLNISWCDQVTKDGIQALVRGCGGLKALFLKGCTQLEDEALKYIGAHCPELVTLNLQTCLQITDEGLITICRGCHKLQSLCASGCSNITDAILNALGQNCPRLRILEVARCSQLTDVGFTTLARNCHELEKMDLEECVQITDSTLIQLSIHCPRLQVLSLSHCELITDDGIRHLGNGACAHDQLEVIELDNCPLITDASLEHLKSCHSLERIELYDCQQITRAGIKRLRTHLPNIKVHAYFAPVTPPPSVGGSRQRFCRCCIIL 3218F38C02EA1BCE ubb_human Polyubiquitin-B UBB protein protein human Homo sapiens Human MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGMQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGGC 33011162F1C48BB1 rims1_mouse Regulating synaptic membrane exocytosis protein 1 Rab-3-interacting molecule 1 Rab-3-interacting protein 1 Rims1 Rab3ip1 Rim1 Kiaa0340 protein protein mouse Mus musculus Mouse MSSAVGPRGPRPPTVPPPMQELPDLSHLTEEERNIIMAVMDRQKEEEEKEEAMLKCVVRDMAKPAACKTPRNAESQPHQPPLNIFRCVCVPRKPSSEEGGPDRNWRLHQQFESYKEQVRKIGEEARRYQGEHKDDAPTCGICHKTKFADGCGHLCSYCRTKFCARCGGRVSLRSNNEDKVVMWVCNLCRKQQEILTKSGAWFFGSGPQQPSQDGTLSDTATGAGSEVPREKKARLQERSRSQTPLSTAAVSSQDTASHGAPLDRNKGAEPSQQALGPEQKQASRSRSEPPRERKKAPGLSEQNGKGGQKSERKRVPKSVVQPGEGTADERERKERRETRRLEKGRSQDYPDRLEKREDGRVAEDEKQRKEEEGVSTPEYTSCEDVELESESVSEKGDLDYWLDPATWHSRETSPISSHPVTWQPSKEGDRLIGRVILNKRTTMPKESGALLGLKVVGGKMTDLGRLGAFITKVKKGSLADVVGHLRAGDEVLEWNGKPLPGATNEEVYNIILESKSEPQVEIIVSRPIGDIPRIPESSHPPLESSSSSFESQKMERPSISVISPTSPGALKDAPQVLPGQLSVKLWYDKVGHQLIVNVLQATDLPPRVDGRPRNPYVKMYFLPDRSDKSKRRTKTVKKLLEPKWNQTFVYSHVHRRDFRERMLEITVWDQPRVQDEESEFLGEILIELETALLDDEPHWYKLQTHDESSLPLPQPSPFMPRRHIHGESSSKKLQRSQRISDSDISDYEVDDGIGVVPPVGYRASARESKATTLTVPEQQRTTHHRSRSVSPHRGDDQGRPRSRLPNVPLQRSLDEIHPTRRSRSPTRHHDASRSLADHRSRHAESQYSSEPDSELLMLPRAKRGRSAECLHMTSELQPSLDRARSASTNCLRPDTSLHSPERERGRWSPSLARRRPASPRIQIQHASPENDRHSRKSERSSIQKQSRKGTASDADRVLPPCLSRRGYAIPRATDQPVIRGKHTTRSRSSEHSSIRTLCSMHHLAPGGSAPPSPLLTRTHRQGSPTQSPPADTSFGSRRGRQLPQVPVRSGSIEQASLVVEERTRQMKMKVHRFKQTTGSGSSQELDHEQYSKYNIHKDQYRSCDNASAKSSDSDVSDVSAISRASSTSRLSSTSFMSEQSERPRGRISSFTPKMQGRRMGTSGRAIIKSTSVSGEIYTLEHNDGSQSDTAVGTVGAGGKKRRSSLSAKVVAIVSRRSRSTSQLSQTESGHKKLKSTIQRSTETGMAAEMRKMVRQPSRESTDGSINSYSSEGNLIFPGVRVGPDSQFSDFLDGLGPAQLVGRQTLATPAMGDIQIGMEDKKGQLEVEVIRARSLTQKPGSKSTPAPYVKVYLLENGACIAKKKTRIARKTLDPLYQQSLVFDESPQGKVLQVIVWGDYGRMDHKCFMGVAQILLEELDLSSMVIGWYKLFPPSSLVDPTLTPLTRRASQSSLESSSGPPCIRS 1E0BE5D522924738 syph_mouse Synaptophysin Major synaptic vesicle protein p38 BM89 antigen Syp protein protein mouse Mus musculus Mouse MLLLADMDVVNQLVAGGQFRVVKEPLGFVKVLQWVFAIFAFATCGSYTGELRLSVECANKTESALNIEVEFEYPFRLHQVYFDAPSCVKGGTTKIFLVGDYSSSAEFFVTVAVFAFLYSMGALATYIFLQNKYRENNKGPMMDFLATAVFAFMWLVSSSAWAKGLSDVKMATDPENIIKEMPMCRQTGNTCKELRDPVTSGLNTSVVFGFLNLVLWVGNLWFVFKETGWAAPFMRAPPGAPEKQPAPGDAYGDAGYGQGPGGYGPQDSYGPQGGYQPDYGQPASGGGGGYGPQGDYGQQGYGQQGAPTSFSNQM F6AC6E30AECE10CE skp1_mouse S-phase kinase-associated protein 1 Skp1 Skp1a S-phase kinase-associated protein 1A Cyclin-A/CDK2-associated protein p19 p19skp1 p19A protein protein mouse Mus musculus Mouse MPTIKLQSSDGEIFEVDVEIAKQSVTIKTMLEDLGMDDEGDDDPVPLPNVNAAILKKVIQWCTHHKDDPPPPEDDENKEKRTDDIPVWDQEFLKVDQGTLFELILAANYLDIKGLLDVTCKTVANMIKGKTPEEIRKTFNIKNDFTEEEEAQVRKENQWCEEK DIP protein Q9WTX5 original sequence version: 100 DIP protein Q9WTX5 original sequence version: 108 AE184C00BE6556DB fbxl20-skp1-1 1 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey association association Similarly, SCRAPPER formed an SCF complex with Skp1 and Cullin1 in mouse brain lysates (Figure 2A). Figure 2A fxl20-rps27a-1 3 6 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Consistent with a function as a ubiquitin ligase, ubiquitinated proteins coimmunoprecipitated (co-IPed) with FLAG-tagged SCRAPPER (FLAG-SCR) from lysates of MG132-treated (a proteasome inhibitor) cells (Figure 2B). Figure 2B fxl20-rps27a-2 4 9 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component colocalization colocalization Immunofluorescence analysis revealed a punctate distribution of SCRAPPER (Figure 1F) that predominantly colocalized with synaptophysin, a known synaptic vesicle protein and presynapse marker. Figure 1F fbxl20-rims1-1 1 6 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Endogenous RIM1 was coimmunoprecipitated with SCRAPPER. Figure 2C fbxl20-rims1-3 4 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization SCRAPPER partially colocalized with RIM1 in cultured neurons (Figure 2D) Figure 2D fbxl20-rims1-2 2 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey c2 region sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) physical association physical association SCRAPPER interacted with the C2B domain of RIM1. Figure 2E fbxl20-rims1-4 2 8 unspecified role unspecified role bait bait c2 region sufficient to bind sufficient binding region sufficient to bind range range range range region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association SCRAPPER interacted with the C2B domain of RIM1. Figure 2E