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/ Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Cancer Cell (1535-6108) 2014 imex curation 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com 20-Oct-2014: Contacted by IntAct-Help. Only protein-protein interactions 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. Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Accepted 2014-SEP-24 by ORCHARD Cancer Cell (1535-6108) 2014 imex curation Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Cancer Cell (1535-6108) 2014 imex curation 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com 20-Oct-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? AMG 655 was detected by anti-human IgG. The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Accepted 2014-SEP-24 by ORCHARD Cancer Cell (1535-6108) 2014 imex curation Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Cancer Cell (1535-6108) 2014 imex curation 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com 20-Oct-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Accepted 2014-SEP-24 by ORCHARD Cancer Cell (1535-6108) 2014 imex curation Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Cancer Cell (1535-6108) 2014 imex curation 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com 20-Oct-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? 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 Accepted 2014-SEP-24 by ORCHARD Cancer Cell (1535-6108) 2014 imex curation Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Apo2L/TRAIL and the Death Receptor 5 Agonist Antibody AMG 655 Cooperate to Promote Receptor Clustering and Antitumor Activity. Cancer Cell (1535-6108) 2014 imex curation 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com 20-Oct-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Fig 3E not added as only a cell lysate used. Cannot find the myeloma line/expression system used (in SLAM) to produce the humanised mAb AMG655- it must be Human? The authors have confirmed that Full length Apo2L/TRAIL is of Human origin. hollandp@amgen.com Accepted 2014-SEP-24 by ORCHARD Cancer Cell (1535-6108) 2014 imex curation 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 60358EAF2A835870 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 DIP protein P50591 original sequence version: 150 DDAAAF78DAAB2F6D 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_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 DIP protein Q13158 original sequence version: 155 DIP protein Q13158 original sequence version: 130 DIP protein Q13158 original sequence version: 110 DIP protein Q13158 original sequence version: 118 DIP protein Q13158 original sequence version: 141 DIP protein Q13158 original sequence version: 166 0E65E2F852E83507 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 DIP protein Q14790 original sequence version: 184 DIP protein Q14790 original sequence version: 135 DIP protein Q14790 original sequence version: 136 DIP protein Q14790 original sequence version: 147 DIP protein Q14790 original sequence version: 146 DIP protein Q14790 original sequence version: 170 DIP protein Q14790 original sequence version: 198 DIP protein Q14790-1 original sequence version: 197 DIP protein Q14790 original sequence version: 158 7A5FEAA6B39B582F 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 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). conatumumab_amg_655-tnfrsf10b-1 4 6 unspecified role unspecified role prey prey human-colo 205 human-colo 205 colo_205 Human colo 205 cells 7 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait association association 3B conatumumab_amg_655-tnfrsf10b-2 4 6 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait physical association physical association 3Bi fadd-casp8-12 3 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey association association 3Ci Cleavage products of Caspase 8 were detected. Apo2L/TRAIL in combination with AMG 655 fadd-casp8-13 3 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 3Cii fadd-casp8-14 5 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey association association 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 5 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association 3Ciii casp8-conatumumab_amg_655-1 2 6 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component 12 unspecified role unspecified role neutral component neutral component colocalization colocalization 3D fab_heavy_chain_amg655-fab_light_chain_amg655 1 11 unspecified role unspecified role neutral component neutral component 6 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 partial_ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component association association Table1 3.3A Working: 22.9%, Free: 28.6% fadd-casp8-17 3 6 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey association association Supp 3C Cleavage products of Caspase 8 were detected. Apo2L/TRAIL in combination with AMG 655