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 amyotrophic lateral sclerosis 8 protein VAPB is cleaved, secreted, and acts as a ligand for Eph receptors. The amyotrophic lateral sclerosis 8 protein VAPB is cleaved, secreted, and acts as a ligand for Eph receptors. Cell (0092-8674) 2008 imex curation Tsuda H., Han SM., Yang Y., Tong C., Lin YQ., Mohan K., Haueter C., Zoghbi A., Harati Y., Kwan J., Miller MA., Bellen HJ. Synapse - Interactions of proteins with an established role in the presynapse. hbellen@bcm.tmc.edu 27-Aug-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Tsuda H., Han SM., Yang Y., Tong C., Lin YQ., Mohan K., Haueter C., Zoghbi A., Harati Y., Kwan J., Miller MA., Bellen HJ. Synapse - Interactions of proteins with an established role in the presynapse. hbellen@bcm.tmc.edu Accepted 2008-AUG-16 by CDEROW. Cell (0092-8674) 2008 Only protein-protein interactions 5 imex curation The amyotrophic lateral sclerosis 8 protein VAPB is cleaved, secreted, and acts as a ligand for Eph receptors. The amyotrophic lateral sclerosis 8 protein VAPB is cleaved, secreted, and acts as a ligand for Eph receptors. Cell (0092-8674) 2008 imex curation Tsuda H., Han SM., Yang Y., Tong C., Lin YQ., Mohan K., Haueter C., Zoghbi A., Harati Y., Kwan J., Miller MA., Bellen HJ. Synapse - Interactions of proteins with an established role in the presynapse. hbellen@bcm.tmc.edu 27-Aug-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro competition binding competition binding anti tag western anti tag western blot anti tag western Tsuda H., Han SM., Yang Y., Tong C., Lin YQ., Mohan K., Haueter C., Zoghbi A., Harati Y., Kwan J., Miller MA., Bellen HJ. Synapse - Interactions of proteins with an established role in the presynapse. hbellen@bcm.tmc.edu Accepted 2008-AUG-16 by CDEROW. 27-Aug-2008: Contacted by JYOTI. Cell (0092-8674) 2008 Only protein-protein interactions 5 imex curation epha4_mouse Ephrin type-A receptor 4 Tyrosine-protein kinase receptor SEK-1 Tyrosine-protein kinase receptor MPK-3 Epha4 Sek Sek1 protein protein mouse Mus musculus Mouse MAGIFYFILFSFLFGICDAVTGSRVYPANEVTLLDSRSVQGELGWIASPLEGGWEEVSIMDEKNTPIRTYQVCNVMEASQNNWLRTDWITREGAQRVYIEIKFTLRDCNSLPGVMGTCKETFNLYYYESDNDKERFIRESQFGKIDTIAADESFTQVDIGDRIMKLNTEIRDVGPLSKKGFYLAFQDVGACIALVSVRVFYKKCPLTVRNLAQFPDTITGADTSSLVEVRGSCVNNSEEKDVPKMYCGADGEWLVPIGNCLCNAGHEEQNGECQACKIGYYKALSTDASCAKCPPHSYSVWEGATSCTCDRGFFRADNDAASMPCTRPPSAPLNLISNVNETSVNLEWSSPQNTGGRQDISYNVVCKKCGAGDPSKCRPCGSGVHYTPQQNGLKTTRVSITDLLAHTNYTFEIWAVNGVSKYNPSPDQSVSVTVTTNQAAPSSIALVQAKEVTRYSVALAWLEPDRPNGVILEYEVKYYEKDQNERSYRIVRTAARNTDIKGLNPLTSYVFHVRARTAAGYGDFSEPLEVTTNTVPSRIIGDGANSTVLLVSVSGSVVLVVILIAAFVISRRRSKYSKAKQEADEEKHLNQGVRTYVDPFTYEDPNQAVREFAKEIDASCIKIEKVIGVGEFGEVCSGRLKVPGKREICVAIKTLKAGYTDKQRRDFLSEASIMGQFDHPNIIHLEGVVTKCKPVMIITEYMENGSLDAFLRKNDGRFTVIQLVGMLRGIGSGMKYLSDMSYVHRDLAARNILVNSNLVCKVSDFGMSRVLEDDPEAAYTTRGGKIPIRWTAPEAIAYRKFTSASDVWSYGIVMWEVMSYGERPYWDMSNQDVIKAIEEGYRLPPPMDCPIALHQLMLDCWQKERSDRPKFGQIVNMLDKLIRNPNSLKRTGSESSRPNTALLDPSSPEFSAVVSVGDWLQAIKMDRYKDNFTAAGYTTLEAVVHMSQDDLARIGITAITHQNKILSSVQAMRTQMQQMHGRMVPV DIP protein Q03137 original sequence version: 161 DIP protein Q03137 original sequence version: 101 89BEB2C7CDB54A55 efnb2_mouse Ephrin-B2 Elf2 Epl5 Eplg5 Htkl Lerk5 Efnb2 EPH-related receptor tyrosine kinase ligand 5 ELF-2 HTK ligand protein protein mouse Mus musculus Mouse MAMARSRRDSVWKYCWGLLMVLCRTAISRSIVLEPIYWNSSNSKFLPGQGLVLYPQIGDKLDIICPKVDSKTVGQYEYYKVYMVDKDQADRCTIKKENTPLLNCARPDQDVKFTIKFQEFSPNLWGLEFQKNKDYYIISTSNGSLEGLDNQEGGVCQTRAMKILMKVGQDASSAGSARNHGPTRRPELEAGTNGRSSTTSPFVKPNPGSSTDGNSAGHSGNNLLGSEVALFAGIASGCIIFIVIIITLVVLLLKYRRRHRKHSPQHTTTLSLSTLATPKRGGNNNGSEPSDVIIPLRTADSVFCPHYEKVSGDYGHPVYIVQEMPPQSPANIYYKV DIP protein P52800 original sequence version: 59 DIP protein P52800 original sequence version: 98 DIP protein P52800 original sequence version: 118 D08894996E399554 vab1_caeel Ephrin receptor 1 vab-1 M03A1.1 Tyrosine-protein kinase Eph receptor Variable abnormal protein 1 protein protein caeel Caenorhabditis elegans MRLYNSRILNPHQSIFILVLQCLITIVTSHQEVLFDLSKVGSDLKWDQVSLRHDRDDVWMEETWRNPAATDEKHANQRAYVTCNYDMINPSNWLFSHFIEVKTARRIYIELLFNTRDCDAYLNPKSCKETFSVYLKQFKTSRPGSTKIEKERFSEDIDNWKNIGRLARSNSNMTTETLGMEIDSDTKTIRIAFEEQGICLSLLNVKIYYRICDEFTDQLVYFRPQVTGPKETDMVRMNGSCIPNASKKIPGVDLIGLCMSTGSGIKTSGECVCDSGYSQIADSNGARCESCPTNTYKPKGQSLCKSCPSNSISSEAASSCRCLNGYFRAEDELISMPCTQPPSRPIKLVANAITATSTRLSWNEPSSLGGRPEIWYEVKCSGRGECGTVVMTPGDKKLSTRSVQINGLRPSSDYTFLVFAKNKVSAQFPEFSEKNAVIDIRTRSEEEDVPPVSHLRVDASQSDGITIAWSVSDSDVSDFEVEVRPAIVKKRTFETRHVNMTYTTFIGLNPETVYQFRVRIRDDLRWSQPISYQLGRGLMSSPSSNEVEESQFLNQTGSALLIIIALILIVIAVALCMIVVQKKSKNRKQMSDLDVLDTYKQDSMTPDYHTTSRHHHHQGNLPATLHEQLRSTTKLNAPLIPSFGSPISQPPPYYGGVHPNSGKYKTYVDPTTYEDPYQALIEFTFDISPNDVFITQVIGGGEFGDVCLGGLSKNSPAAAKWSVSNTTMGRGGGGGGYESEPYETVAIKTLKSGSSAKAKAEFLTEATIMGQFSHPNVIRLIGVVTSAEPVMIVAEYMANGSLDQFLRNTDQRGEKVHWEKITEMLYGIASGMKYLTDMGYVHRVSFLRDLAARNVLLDMELRCKIADFGLSRGVRSEGSSVEPEYTTNGGKIPVRWTAPEAITHRKFTPSSDVWSFGVVIWEVCSFGERPYWDWTNQKVISEVMIGYRLPPPMDCPMGLYRIAQWCWKMERHERPTFTQLLATFHKYILQPTLIEHDPGELPRRVQSQSALNTYGSVNVGVVPTPPSSAAPMPSLDDFLRQIGLNHVYGQLVSNNIHSVSDLANTSHLDLLAYGLMSAECSTVRDGLNGRISGSPPGSSGTIHATTRGTRTTRPPREEGFFV 6E4A3037BB92A1D0 o44782_caeel Major sperm protein vpr-1 CELE_F33D11.11 F33D11.11 protein protein caeel Caenorhabditis elegans MSEKHSLLQVTPNRELVFTGPFSDVVTSHMTLKNTSTNPVCFKVKTTAPKQYCVRPNSGLLKSGDSKQITVMLQPLEGIPSDAGRHKFMVQSCVAPSEDLPDLESVWKIIDPAELTYSKLMVTFVDKRDPASGDDTKTFAVRNEDSFASSGQAQELGSSYSGTPSHDGTVNSLRKSLKSTVDEKEELQKKVHGLEQEIEVMLKKNRKLQQSHSDGALVDGAYPTLQIFLIAVAALLIGLIVGRLF 5619C7921986991E vapb_human Vesicle-associated membrane protein-associated protein B/C VAPB UNQ484/PRO983 protein protein human Homo sapiens Human MAKVEQVLSLEPQHELKFRGPFTDVVTTNLKLGNPTDRNVCFKVKTTAPRRYCVRPNSGIIDAGASINVSVMLQPFDYDPNEKSKHKFMVQSMFAPTDTSDMEAVWKEAKPEDLMDSKLRCVFELPAENDKPHDVEINKIISTTASKTETPIVSKSLSSSLDDTEVKKVMEECKRLQGEVQRLREENKQFKEEDGLRMRKTVQSNSPISALAPTGKEEGLSTRLLALVVLFFIVGVIIGKIAL DIP protein O95292 original sequence version: 141 22AEEF9EC7FC0B3F vpr1-vab1-1 1 5 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 extracellular region sufficient to bind sufficient binding region sufficient to bind range range range range 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. 6 unspecified role unspecified role bait bait msp region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 13 11 direct interaction direct interaction Purified FLAG-tagged VPR-1 MSP (VPR-1MSP-FLAG) and V5-tagged VAB-1 ECT domain (VAB-1 Ex-V5) were coimmunoprecipitated using FLAG antibody (C) and V5 antibody (D) Fig. 7C vab-1 (O61440) domain boundaries taken from UniprotKB vpr1-vab1-2 1 5 unspecified role unspecified role bait bait extracellular region sufficient to bind sufficient binding region sufficient to bind range range range range region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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. 6 unspecified role unspecified role prey prey 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 msp region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Purified FLAG-tagged VPR-1 MSP (VPR-1MSP-FLAG) and V5-tagged VAB-1 ECT domain (VAB-1 Ex-V5) were coimmunoprecipitated using FLAG antibody (C) and V5 antibody (D) Fig. 7D vab-1 (O61440) domain boundaries taken from UniprotKB epha4-vapb-1 1 7 unspecified role unspecified role prey prey msp region sufficient to bind sufficient binding region sufficient to bind range range range range region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 3 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ectodomain sufficient to bind sufficient binding region sufficient to bind range range range range 28 24 physical association physical association Where exact lengths of construct were not given, domain boundaries have been taken from the UniProt records We incubated mEphA4Ex-V5His together with hVAPMSP-His proteins and performed coIP assays with the V5 antibody. Fig. 7E expression details of mammalian proteins not given epha4-efnb2-1 1 3 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 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. 4 unspecified role unspecified role prey prey fc tagged region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association After incubation, we retrieved mEphrinB2-Fc and EphA4 proteins complexes and immunoblotted with the anti-FLAG antibody. Fig. 7F, 7G Conditioned medium containing hVAP (O95292, MSP domain construct) disrupts the interaction epha4-vapb-2 2 4 inhibitor inhibitor prey prey 3 unspecified role unspecified role bait bait region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ectodomain sufficient to bind sufficient binding region sufficient to bind range range range range 7 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined msp region sufficient to bind sufficient binding region sufficient to bind range range range range association association Where exact lengths of construct were not given, domain boundaries have been taken from the UniProt records To confirm competition, we incubated mEphA4Ex-V5His and hVAPMSP-His proteins together with EphrinB2 protein. Fig. 7H, 7I Expression details of mammalian proteins not given EphrinB2 is able to outcompete hVAPB MSP for binding to EphA4