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/ Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. J. Biol. Chem. (0021-9258) 2006 imex curation Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro competition binding competition binding predetermined predetermined participant predetermined Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg Accepted 2006-SEP-11 by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. J. Biol. Chem. (0021-9258) 2006 imex curation Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro biochemical biochemical predetermined predetermined participant predetermined Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology It was assumed that the alpha beta integrin used in this experiment was human as this is the purified integrin supplied by Chemicon. The organism origin of the alpha beta integrin was not stated buy the authors. 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg Accepted 2006-SEP-11 by JYOTI. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. J. Biol. Chem. (0021-9258) 2006 imex curation Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-cs Homo sapiens fibroblast from patients with Cockayne syndrome cs Fibroblast from patients with Cockayne syndrome transformed with SV40 anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg Accepted 2006-SEP-11 by JYOTI. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin. J. Biol. Chem. (0021-9258) 2006 imex curation Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro 3d-em 3D electron microscopy 3D-EM electron tomog predetermined predetermined participant predetermined Lee JW., Chu JJ., Ng ML. BioCreative - Critical Assessment of Information Extraction systems in Biology It was assumed that the alpha beta integrin used in this experiment was human as this is the purified integrin supplied by Chemicon. The organism origin of the alpha beta integrin was not stated buy the authors. Will request term 'AFM atomic force microscope'. 96-Well plates were coated with Beta 3 integrin (150 ng/well) and incubated with WNV E DIII protein (various concentrations) for 1 h at 37C, before the addition of radiolabeled WNV particles micngml@nus.edu.sg Accepted 2006-SEP-11 by JYOTI. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation polg_wnv Genome polyprotein protein protein wnv West Nile virus (WNV) MSKKPGGPGKNRAVNMLKRGMPRGLSLIGLKRAMLSLIDGKGPIRFVLALLAFFRFTAIAPTRAVLDRWRGVNKQTAMKHLLSFKKELGTLTSAINRRSTKQKKRGGTAGFTILLGLIACAGAVTLSNFQGKVMMTVNATDVTDVITIPTAAGKNLCIVRAMDVGYLCEDTITYECPVLAAGNDPEDIDCWCTKSSVYVRYGRCTKTRHSRRSRRSLTVQTHGESTLANKKGAWLDSTKATRYLVKTESWILRNPGYALVAAVIGWMLGSNTMQRVVFAILLLLVAPAYSFNCLGMSNRDFLEGVSGATWVDLVLEGDSCVTIMSKDKPTIDVKMMNMEAANLADVRSYCYLASVSDLSTRAACPTMGEAHNEKRADPAFVCKQGVVDRGWGNGCGLFGKGSIDTCAKFACTTKATGWIIQKENIKYEVAIFVHGPTTVESHGKIGATQAGRFSITPSAPSYTLKLGEYGEVTVDCEPRSGIDTSAYYVMSVGEKSFLVHREWFMDLNLPWSSAGSTTWRNRETLMEFEEPHATKQSVVALGSQEGALHQALAGAIPVEFSSNTVKLTSGHLKCRVKMEKLQLKGTTYGVCSKAFKFARTPADTGHGTVVLELQYTGTDGPCKVPISSVASLNDLTPVGRLVTVNPFVSVATANSKVLIELEPPFGDSYIVVGRGEQQINHHWHKSGSSIGKAFTTTLRGAQRLAALGDTAWDFGSVGGVFTSVGKAIHQVFGGAFRSLFGGMSWITQGLLGALLLWMGINARDRSIAMTFLAVGGVLLFLSVNVHADTGCAIDIGRQELRCGSGVFIHNDVEAWMDRYKFYPETPQGLAKIIQKAHAEGVCGLRSVSRLEHQMWEAIKDELNTLLKENGVDLSVVVEKQNGMYKAAPKRLAATTEKLEMGWKAWGKSIIFAPELANNTFVIDGPETEECPTANRAWNSMEVEDFGFGLTSTRMFLRIRETNTTECDSKIIGTAVKNNMAVHSDLSYWIESGLNDTWKLERAVLGEVKSCTWPETHTLWGDGVLESDLIIPITLAGPRSNHNRRPGYKTQNQGPWDEGRVEIDFDYCPGTTVTISDSCEHRGPAARTTTESGKLITDWCCRSCTLPPLRFQTENGCWYGMEIRPTRHDEKTLVQSRVNAYNADMIDPFQLGLMVVFLATQEVLRKRWTAKISIPAIMLALLVLVFGGITYTDVLRYVILVGAAFAEANSGGDVVHLALMATFKIQPVFLVASFLKARWTNQESILLMLAAAFFQMAYYDAKNVLSWEVPDVLNSLSVAWMILRAISFTNTSNVVVPLLALLTPGLKCLNLDVYRILLLMVGVGSLIKEKRSSAAKKKGACLICLALASTGVFNPMILAAGLMACDPNRKRGWPATEVMTAVGLMFAIVGGLAELDIDSMAIPMTIAGLMFAAFVISGKSTDMWIERTADITWESDAEITGSSERVDVRLDDDGNFQLMNDPGAPWKIWMLRMACLAISAYTPWAILPSVIGFWITLQYTKRGGVLWDTPSPKEYKKGDTTTGVYRIMTRGLLGSYQAGAGVMVEGVFHTLWHTTKGAALMSGEGRLDPYWGSVKEDRLCYGGPWKLQHKWNGHDEVQMIVVEPGKNVKNVQTKPGVFKTPEGEIGAVTLDYPTGTSGSPIVDKNGDVIGLYGNGVIMPNGSYISAIVQGERMEEPAPAGFEPEMLRKKQITVLDLHPGAGKTRKILPQIIKEAINKRLRTAVLAPTRVVAAEMSEALRGLPIRYQTSAVHREHSGNEIVDVMCHATLTHRLMSPHRVPNYNLFIMDEAHFTDPASIAARGYIATKVELGEAAAIFMTATPPGTSDPFPESNAPISDMQTEIPDRAWNTGYEWITEYVGKTVWFVPSVKMGNEIALCLQRAGKKVIQLNRKSYETEYPKCKNDDWDFVITTDISEMGANFKASRVIDSRKSVKPTIIEEGDGRVILGEPSAITAASAAQRRGRIGRNPSQVGDEYCYGGHTNEDDSNFAHWTEARIMLDNINMPNGLVAQLYQPEREKVYTMDGEYRLRGEERKNFLEFLRTADLPVWLAYKVAAAGISYHDRKWCFDGPRTNTILEDNNEVEVITKLGERKILRPRWADARVYSDHQALKSFKDFASGKRSQIGLVEVLGRMPEHFMVKTWEALDTMYVVATAEKGGRAHRMALEELPDALQTIVLIALLSVMSLGVFFLLMQRKGIGKIGLGGVILGAATFFCWMAEVPGTKIAGMLLLSLLLMIVLIPEPEKQRSQTDNQLAVFLICVLTLVGAVAANEMGWLDKTKNDIGSLLGHRPEARETTLGVESFLLDLRPATAWSLYAVTTAVLTPLLKHLITSDYINTSLTSINVQASALFTLARGFPFVDVGVSALLLAVGCWGQVTLTVTVTAAALLFCHYAYMVPGWQAEAMRSAQRRTAAGIMKNVVVDGIVATDVPELERTTPVMQKKVGQIILILVSMAAVVVNPSVRTVREAGILTTAAAVTLWENGASSVWNATTAIGLCHIMRGGWLSCLSIMWTLIKNMEKPGLKRGGAKGRTLGEVWKERLNHMTKEEFTRYRKEAITEVDRSAAKHARREGNITGGHPVSRGTAKLRWLVERRFLEPVGKVVDLGCGRGGWCYYMATQKRVQEVKGYTKGGPGHEEPQLVQSYGWNIVTMKSGVDVFYRPSEASDTLLCDIGESSSSAEVEEHRTVRVLEMVEDWLHRGPKEFCIKVLCPYMPKVIEKMETLQRRYGGGLIRNPLSRNSTHEMYWVSHASGNIVHSVNMTSQVLLGRMEKKTWKGPQFEEDVNLGSGTRAVGKPLLNSDTSKIKNRIERLKKEYSSTWHQDANHPYRTWNYHGSYEVKPTGSASSLVNGVVRLLSKPWDTITNVTTMAMTDTTPFGQQRVFKEKVDTKAPEPPEGVKYVLNETTNWLWAFLARDKKPRMCSREEFIGKVNSNAALGAMFEEQNQWKNAREAVEDPKFWEMVDEEREAHLRGECNTCIYNMMGKREKKPGEFGKAKGSRAIWFMWLGARFLEFEALGFLNEDHWLGRKNSGGGVEGLGLQKLGYILKEVGTKPGGKVYADDTAGWDTRITKADLENEAKVLELLDGEHRRLARSIIELTYRHKVVKVMRPAADGKTVMDVISREDQRGSGQVVTYALNTFTNLAVQLVRMMEGEGVIGPDDVEKLGKGKGPKVRTWLFENGEERLSRMAVSGDDCVVKPLDDRFATSLHFLNAMSKVRKDIQEWKPSTGWYDWQQVPFCSNHFTELIMKDGRTLVVPCRGQDELIGRARISPGAGWNVRDTACLAKSYAQMWLLLYFHRRDLRLMANAICSAVPANWVPTGRTTWSIHAKGEWMTTEDMLAVWNRVWIEENEWMEDKTPVERWSDVPYSGKREDIWCGSLIGTRTRATWAENIHVAINQVRSVIGEEKYVDYMSSLRRYEDTIVVEDTVL 42D71B7CB12DC45B itb3_human Integrin beta-3 Platelet membrane glycoprotein IIIa ITGB3 GP3A protein protein human Homo sapiens Human MRARPRPRPLWATVLALGALAGVGVGGPNICTTRGVSSCQQCLAVSPMCAWCSDEALPLGSPRCDLKENLLKDNCAPESIEFPVSEARVLEDRPLSDKGSGDSSQVTQVSPQRIALRLRPDDSKNFSIQVRQVEDYPVDIYYLMDLSYSMKDDLWSIQNLGTKLATQMRKLTSNLRIGFGAFVDKPVSPYMYISPPEALENPCYDMKTTCLPMFGYKHVLTLTDQVTRFNEEVKKQSVSRNRDAPEGGFDAIMQATVCDEKIGWRNDASHLLVFTTDAKTHIALDGRLAGIVQPNDGQCHVGSDNHYSASTTMDYPSLGLMTEKLSQKNINLIFAVTENVVNLYQNYSELIPGTTVGVLSMDSSNVLQLIVDAYGKIRSKVELEVRDLPEELSLSFNATCLNNEVIPGLKSCMGLKIGDTVSFSIEAKVRGCPQEKEKSFTIKPVGFKDSLIVQVTFDCDCACQAQAEPNSHRCNNGNGTFECGVCRCGPGWLGSQCECSEEDYRPSQQDECSPREGQPVCSQRGECLCGQCVCHSSDFGKITGKYCECDDFSCVRYKGEMCSGHGQCSCGDCLCDSDWTGYYCNCTTRTDTCMSSNGLLCSGRGKCECGSCVCIQPGSYGDTCEKCPTCPDACTFKKECVECKKFDRGALHDENTCNRYCRDEIESVKELKDTGKDAVNCTYKNEDDCVVRFQYYEDSSGKSILYVVEEPECPKGPDILVVLLSVMGAILLIGLAALLIWKLLITIHDRKEFAKFEEERARAKWDTANNPLYKEATSTFTNITYRGT DIP protein P05106 original sequence version: 202 DIP protein P05106-1 original sequence version: 158 DIP protein P05106-1 original sequence version: 164 DIP protein P05106 original sequence version: 200 DIP protein P05106-1 original sequence version: 213 DIP protein P05106-1 original sequence version: 175 DIP protein P05106-1 original sequence version: 192 DIP protein P05106-1 original sequence version: 171 F246623608E05F9E itgb3-e_protein 1 6 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey region Domain III of E protein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chlae-vero Chlorocebus (Cercopithecus) aethiops kidney cells Grivet vero Chlorocebus (Cercopithecus) aethiops kidney cells similar to cos-7 cells but these are not trasnformed physical association physical association Figure 1 A Dosage-dependent inhibition of virus binding to V3 integrin is observed with increasing concentrations of WNV E DIII proteins. Coordinates for domain III of the WNV E protein were obtained from Rey, F. A., Heinz, F. X., Mandl, C., Kunz, C., & Harrison, S. C. (1995) Nature 375, 291�298 a reference provided by the authors. Recombinant WNV E DIII Protein Interacts with V 3 Integrin itgb3-e_protein-1 3 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region Domain III of E protein 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 physical association physical association Figure 1B Coordinates for domain III of the WNV E protein were obtained from Rey, F. A., Heinz, F. X., Mandl, C., Kunz, C., & Harrison, S. C. (1995) Nature 375, 291�298 a reference provided by the authors. Recombinant WNV E DIII Protein Interacts with V 3 Integrin itgb3-e_protein-2 2 6 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey region Domain III of E protein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chlae-vero Chlorocebus (Cercopithecus) aethiops kidney cells Grivet vero Chlorocebus (Cercopithecus) aethiops kidney cells similar to cos-7 cells but these are not trasnformed physical association physical association Figure 1C Coordinates for domain III of the WNV E protein were obtained from Rey, F. A., Heinz, F. X., Mandl, C., Kunz, C., & Harrison, S. C. (1995) Nature 375, 291�298 a reference provided by the authors. Inhibition of WNV Entry by Soluble WNV E DIII Protein itgb3-e_protein-3 4 5 unspecified role unspecified role neutral component neutral component region Domain III of E protein sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chlae-vero Chlorocebus (Cercopithecus) aethiops kidney cells Grivet vero Chlorocebus (Cercopithecus) aethiops kidney cells similar to cos-7 cells but these are not trasnformed 6 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Figure 3 Figure 7. Coordinates for domain III of the WNV E protein were obtained from Rey, F. A., Heinz, F. X., Mandl, C., Kunz, C., & Harrison, S. C. (1995) Nature 375, 291�298 a reference provided by the authors. Binding force between WNV E DIII protein interactions with the V 3 integrin molecule. A force of 45 pN was measured when interaction between ITGB3 protein and domain III E protein was measured by AFM. Occasionally there were bindings at abnormally large magnitude, (see Fig. 5a).In the example given in the text the force curve was measured to be 83 pN. Thre authors state that these may arise from multiple protein interactions between wafer and probe during AFM analysis. Addition of beta3 integrin to the assay medium inhibited interaction between probe bound beta 3 integrin and wafer bound E protein (domain III).