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/ graves-2014-2 Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. human-colo 205 human-colo 205 colo_205 Human colo 205 cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Graves J.D., Kordich J.J., Huang T.H., Piasecki J., Bush T.L., Sullivan T., Foltz I.N., Chang W., Douangpanya H., Dang T., O'Neill J.W., Mallari R., Zhao X., Branstetter D.G., Rossi J.M., Long A.M., Huang X., Holland P.M. Cancer - Interactions investigated in the context of cancer The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Cancer Cell (1535-6108) 2014 imex curation Only protein-protein interactions graves-2014-1 Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. in vitro In vitro affinity techniques affinity technology affinity techniques western blot western blot Immuno blot Immuno blot Graves J.D., Kordich J.J., Huang T.H., Piasecki J., Bush T.L., Sullivan T., Foltz I.N., Chang W., Douangpanya H., Dang T., O'Neill J.W., Mallari R., Zhao X., Branstetter D.G., Rossi J.M., Long A.M., Huang X., Holland P.M. Cancer - Interactions investigated in the context of cancer Colo-205 cell lysates were treated with exogenous AMG 655 and/or TRAIL, then Protein G was used to pull down AMG-655 and associated proteins. The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Cancer Cell (1535-6108) 2014 imex curation Only protein-protein interactions graves-2014-3 Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. human-sklu1 Human SKLU1 Lung Adenocarcincinoma cell line sklu1 SKLU1 Lung Adenocarcincinoma ATCC HTB57 http://www.lgcstandards-atcc.org/products/all/HTB-57.aspx?geo_country=gb anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Graves J.D., Kordich J.J., Huang T.H., Piasecki J., Bush T.L., Sullivan T., Foltz I.N., Chang W., Douangpanya H., Dang T., O'Neill J.W., Mallari R., Zhao X., Branstetter D.G., Rossi J.M., Long A.M., Huang X., Holland P.M. Cancer - Interactions investigated in the context of cancer The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Cancer Cell (1535-6108) 2014 imex curation Only protein-protein interactions graves-2014-4 Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. human-sklu1 Human SKLU1 Lung Adenocarcincinoma cell line sklu1 SKLU1 Lung Adenocarcincinoma ATCC HTB57 http://www.lgcstandards-atcc.org/products/all/HTB-57.aspx?geo_country=gb confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Graves J.D., Kordich J.J., Huang T.H., Piasecki J., Bush T.L., Sullivan T., Foltz I.N., Chang W., Douangpanya H., Dang T., O'Neill J.W., Mallari R., Zhao X., Branstetter D.G., Rossi J.M., Long A.M., Huang X., Holland P.M. Cancer - Interactions investigated in the context of cancer AMG 655 was detected by anti-human IgG. The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Cancer Cell (1535-6108) 2014 imex curation Only protein-protein interactions graves-2014-5 Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Graves J.D., Kordich J.J., Huang T.H., Piasecki J., Bush T.L., Sullivan T., Foltz I.N., Chang W., Douangpanya H., Dang T., O'Neill J.W., Mallari R., Zhao X., Branstetter D.G., Rossi J.M., Long A.M., Huang X., Holland P.M. Cancer - Interactions investigated in the context of cancer PDBe entry 4n90 gives sequence for the DR5 extracellular domain as aas 57-182, wheras this publication gives it as aas 54-183. The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Cancer Cell (1535-6108) 2014 imex curation Only protein-protein interactions casp8_human Caspase-8 CASP8 MCH5 ICE-like apoptotic protease 5 MORT1-associated ced-3 homolog FADD-homologous ICE/ced-3-like protease FADD-like ICE Apoptotic cysteine protease Apoptotic protease Mch-5 CAP4 protein protein human Homo sapiens Human MDFSRNLYDIGEQLDSEDLASLKFLSLDYIPQRKQEPIKDALMLFQRLQEKRMLEESNLSFLKELLFRINRLDLLITYLNTRKEEMERELQTPGRAQISAYRVMLYQISEEVSRSELRSFKFLLQEEISKCKLDDDMNLLDIFIEMEKRVILGEGKLDILKRVCAQINKSLLKIINDYEEFSKERSSSLEGSPDEFSNGEELCGVMTISDSPREQDSESQTLDKVYQMKSKPRGYCLIINNHNFAKAREKVPKLHSIRDRNGTHLDAGALTTTFEELHFEIKPHDDCTVEQIYEILKIYQLMDHSNMDCFICCILSHGDKGIIYGTDGQEAPIYELTSQFTGLKCPSLAGKPKVFFIQACQGDNYQKGIPVETDSEEQPYLEMDLSSPQTRYIPDEADFLLGMATVNNCVSYRNPAEGTWYIQSLCQSLRERCPRGDDILTILTEVNYEVSNKDDKKNMGKQMPQPTFTLRKKLVFPSD fadd_human FAS-associated death domain protein FAS-associating death domain-containing protein Mediator of receptor induced toxicity FADD MORT1 GIG3 Growth-inhibiting gene 3 protein protein protein human Homo sapiens Human MDPFLVLLHSVSSSLSSSELTELKFLCLGRVGKRKLERVQSGLDLFSMLLEQNDLEPGHTELLRELLASLRRHDLLRRVDDFEAGAAAGAAPGEEDLCAAFNVICDNVGKDWRRLARQLKVSDTKIDSIEDRYPRNLTERVRESLRIWKNTEKENATVAHLVGALRSCQMNLVADLVQEVQQARDLQNRSGAMSPMSWNSDASTSEAS tr10b_human Tumor necrosis factor receptor superfamily member 10B Death receptor 5 TNF-related apoptosis-inducing ligand receptor 2 UNQ160/PRO186 TNFRSF10B DR5 KILLER TRAILR2 TRICK2 ZTNFR9 protein protein human Homo sapiens Human MEQRGQNAPAASGARKRHGPGPREARGARPGPRVPKTLVLVVAAVLLLVSAESALITQQDLAPQQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKHSGEVPAVEETVTSSPGTPASPCSLSGIIIGVTVAAVVLIVAVFVCKSLLWKKVLPYLKGICSGGGGDPERVDRSSQRPGAEDNVLNEIVSILQPTQVPEQEMEVQEPAEPTGVNMLSPGESEHLLEPAEAERSQRRRLLVPANEGDPTETLRQCFDDFADLVPFDSWEPLMRKLGLMDNEIKVAKAEAAGHRDTLYTMLIKWVNKTGRDASVHTLLDALETLGERLAKQKIEDHLLSSGKFMYLEGNADSAMS tnf10_human Tumor necrosis factor ligand superfamily member 10 TNF-related apoptosis-inducing ligand Apo-2 ligand TNFSF10 APO2L TRAIL protein protein human Homo sapiens Human MAMMEVQGGPSLGQTCVLIVIFTVLLQSLCVAVTYVYFTNELKQMQDKYSKSGIACFLKEDDSYWDPNDEESMNSPCWQVKWQLRQLVRKMILRTSEETISTVQEKQQNISPLVRERGPQRVAAHITGTRGRSNTLSSPNSKNEKALGRKINSWESSRSGHSFLSNLHLRNGELVIHEKGFYYIYSQTYFRFQEEIKENTKNDKQMVQYIYKYTSYPDPILLMKSARNSCWSKDAEYGLYSIYQGGIFELKENDRIFVSVTNEHLIDMDHEASFFGAFLVG conatumumab/amg_655 AMG 655/conatumumab protein protein human Homo sapiens Human QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGDYFWSWIRQLPGKGLEWIGHIHNSGTTYYNPSLKSRVTISVDTSKKQFSLRLSSVTAADTAVYYCARDRGGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKCONATUMUMABLIGHTCHAINEIVLTQSPGTLSLSPGERATLSCRASQGISRSYLAWYQQKPGQAPSLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQFGSSPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC A humanised Monoclonal antibody. The sequence entered has been taken from Chembl entry 1743003; one Human IgG1 heavy and one light chain. While no exact sequence match can be found in any protein database, the heavy chain component (aas123-452) has a 100% match with the complete sequence (aas 1-330) from UniProt P01857 -IgG1- C region, (Gene ID IGHG1), and the AMG_655 light chain (aas 110-215) has 100% match with the complete sequence from P01834 (aas 1-106) -Ig kappa chain C region, (Gene ID IGKC). fab_light_chain_amg655 Fab fragment-light chain,AMG 655/conatumumab protein protein human Homo sapiens Human EIVLTQSPGTLSLSPGERATLSCRASQGISRSYLAWYQQKPGQAPSLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQFGSSPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC sequence taken from CHEMBL1743003 from complete light chain sequence given: this appears to be the same sequence as given in PDBe entry 4n90 fab_heavy_chain_amg655 Fab fragment-heavy chain,AMG 655/conatumumab protein protein human Homo sapiens Human QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGDYFWSWIRQLPGKGLEWIGHIHNSGTTYYNPSLKSRVTISVDTSKKQFSLRLSSVTAADTAVYYCARDRGGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS Sequence is given in PDBe entry 4n90. fadd-casp8-12 1566612 n/a 1566617 unspecified role unspecified role prey prey n/a 1566618 unspecified role unspecified role bait bait n/a 1566619 unspecified role unspecified role prey prey association association false false false 3Ci Cleavage products of Caspase 8 were detected. Apo2L/TRAIL in combination with AMG 655 fadd-casp8-13 1566612 n/a 1566618 unspecified role unspecified role bait bait n/a 1566617 unspecified role unspecified role prey prey physical association physical association false false false 3Cii fadd-casp8-17 1566612 n/a 1566619 unspecified role unspecified role prey prey n/a 1566617 unspecified role unspecified role bait bait n/a 1566618 unspecified role unspecified role prey prey association association false false false Supp 3C Cleavage products of Caspase 8 were detected. Apo2L/TRAIL in combination with AMG 655 conatumumab_amg_655-tnfrsf10b-1 1566613 n/a 1566619 unspecified role unspecified role prey prey human-colo 205 human-colo 205 colo_205 Human colo 205 cells n/a 1566620 unspecified role unspecified role prey prey n/a 1566621 unspecified role unspecified role bait bait association association false false false 3B conatumumab_amg_655-tnfrsf10b-2 1566613 n/a 1566619 unspecified role unspecified role prey prey n/a 1566621 unspecified role unspecified role bait bait physical association physical association false false false 3Bi fadd-casp8-14 1566614 n/a 1566619 unspecified role unspecified role prey prey n/a 1566618 unspecified role unspecified role bait bait n/a 1566617 unspecified role unspecified role prey prey association association false false false 3Ci Cleavage products of Caspase 8 were detected. In the absence of AMG-655 the interaction was much weaker. Apo2L/TRAIL in combination with AMG 655 fadd-casp8-16 1566614 n/a 1566617 unspecified role unspecified role prey prey n/a 1566618 unspecified role unspecified role bait bait physical association physical association false false false 3Ciii casp8-conatumumab_amg_655-1 1566615 n/a 1566619 unspecified role unspecified role neutral component neutral component n/a 1566617 unspecified role unspecified role neutral component neutral component n/a 1566621 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 3D fab_heavy_chain_amg655-fab_light_chain_amg655 1566616 n/a 1566622 unspecified role unspecified role neutral component neutral component Stoichiometry: 3.0 n/a 1566619 unspecified role unspecified role neutral component neutral component c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false partial_ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 3.0 n/a 1566620 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 3.0 n/a 1566623 unspecified role unspecified role neutral component neutral component Stoichiometry: 3.0 association association false false false Table1 3.3A Working: 22.9%, Free: 28.6%