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/ An Fcγ receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. An Fcγ receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. Cancer Cell (1535-6108) 2011 imex curation Wilson NS., Yang B., Yang A., Loeser S., Marsters S., Lawrence D., Li Y., Pitti R., Totpal K., Yee S., Ross S., Vernes JM., Lu Y., Adams C., Offringa R., Kelley B., Hymowitz S., Daniel D., Meng G., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis Rag2−/− mice (no mature B or T lymphocytes and SCID phenotype) with Colo205 tumors. aa@gene.com 06-Jun-2011: Contacted by JYOTI. jyoti Only protein-protein interactions mouse Mus musculus Mouse anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wilson NS., Yang B., Yang A., Loeser S., Marsters S., Lawrence D., Li Y., Pitti R., Totpal K., Yee S., Ross S., Vernes JM., Lu Y., Adams C., Offringa R., Kelley B., Hymowitz S., Daniel D., Meng G., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis Rag2−/− mice (no mature B or T lymphocytes and SCID phenotype) with Colo205 tumors. aa@gene.com Accepted 2011-FEB-18 by JYOTI 06-Jun-2011: Contacted by JYOTI. Cancer Cell (1535-6108) 2011 jyoti Only protein-protein interactions imex curation An Fcγ receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. An Fcγ receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. Cancer Cell (1535-6108) 2011 imex curation Wilson NS., Yang B., Yang A., Loeser S., Marsters S., Lawrence D., Li Y., Pitti R., Totpal K., Yee S., Ross S., Vernes JM., Lu Y., Adams C., Offringa R., Kelley B., Hymowitz S., Daniel D., Meng G., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis Rag2−/− mice (no mature B or T lymphocytes and SCID phenotype) with Colo205 tumors. aa@gene.com 06-Jun-2011: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa predetermined predetermined participant predetermined Wilson NS., Yang B., Yang A., Loeser S., Marsters S., Lawrence D., Li Y., Pitti R., Totpal K., Yee S., Ross S., Vernes JM., Lu Y., Adams C., Offringa R., Kelley B., Hymowitz S., Daniel D., Meng G., Ashkenazi A. Apoptosis - Interactions involving proteins with a function related to apoptosis mgt has contacted the authors and not had any response Rag2−/− mice (no mature B or T lymphocytes and SCID phenotype) with Colo205 tumors. details for R2A/3A were obtained from Shileds -2001. Genbank BC027310 (UniProt Q8R2R4) - is the receptor 4 type, FcγRIV - Mancardi-2008. aa@gene.com Accepted 2011-FEB-18 by JYOTI 06-Jun-2011: Contacted by JYOTI. Cancer Cell (1535-6108) 2011 jyoti Only protein-protein interactions imex curation fcgr3_mouse Low affinity immunoglobulin gamma Fc region receptor III Fcgr3 Fc-gamma RIII protein protein mouse Mus musculus Mouse MFQNAHSGSQWLLPPLTILLLFAFADRQSAALPKAVVKLDPPWIQVLKEDMVTLMCEGTHNPGNSSTQWFHNGRSIRSQVQASYTFKATVNDSGEYRCQMEQTRLSDPVDLGVISDWLLLQTPQRVFLEGETITLRCHSWRNKLLNRISFFHNEKSVRYHHYKSNFSIPKANHSHSGDYYCKGSLGSTQHQSKPVTITVQDPATTSSISLVWYHTAFSLVMCLLFAVDTGLYFYVRRNLQTPREYWRKSLSIRKHQAPQDK 757FB83668E41069 fcgr2_mouse Low affinity immunoglobulin gamma Fc region receptor II Fcgr2 Fcgr2b Ly-17 Fc gamma receptor IIB IgG Fc receptor II beta Lymphocyte antigen 17 protein protein mouse Mus musculus Mouse MESNWTVHVFSRTLCHMLLWTAVLNLAAGTHDLPKAVVKLEPPWIQVLKEDTVTLTCEGTHNPGNSSTQWFHNGRSIRSQVQASYTFKATVNDSGEYRCQMEQTRLSDPVDLGVISDWLLLQTPQLVFLEGETITLRCHSWRNKLLNRISFFHNEKSVRYHHYSSNFSIPKANHSHSGDYYCKGSLGRTLHQSKPVTITVQGPKSSRSLPVLTIVAAVTGIAVAAIVIILVSLVYLKKKQVPALPGNPDHREMGETLPEEVGEYRQPSGGSVPVSPGPPSGLEPTSSSPYNPPDLEEAAKTEAENTITYSLLKHPEALDEETEHDYQNHI A463B9EFF2717511 drozitumab drozitumab-anti-DR5(CD262)Human IgG1 heavy chain, disulfide linked to human λ3-chain, dimer. protein protein human Homo sapiens Human fcgr4_mouse Low affinity immunoglobulin gamma Fc region receptor IV Fcgr4 CD16-2 FcgammaRIV protein protein mouse Mus musculus Mouse MWQLLLPTALVLTAFSGIQAGLQKAVVNLDPKWVRVLEEDSVTLRCQGTFSPEDNSIKWFHNESLIPHQDANYVIQSARVKDSGMYRCQTALSTISDPVQLEVHMGWLLLQTTKWLFQEGDPIHLRCHSWQNRPVRKVTYLQNGKGKKYFHENSELLIPKATHNDSGSYFCRGLIGHNNKSSASFRISLGDPGSPSMFPPWHQITFCLLIGLLFAIDTVLYFSVRRGLQSPVADYEEPKIQWSKEPQDK 99BBA7992862E80C fcgr1_mouse High affinity immunoglobulin gamma Fc receptor I Fcgr1 Fcg1 Fc-gamma RI protein protein mouse Mus musculus Mouse MILTSFGDDMWLLTTLLLWVPVGGEVVNATKAVITLQPPWVSIFQKENVTLWCEGPHLPGDSSTQWFINGTAVQISTPSYSIPEASFQDSGEYRCQIGSSMPSDPVQLQIHNDWLLLQASRRVLTEGEPLALRCHGWKNKLVYNVVFYRNGKSFQFSSDSEVAILKTNLSHSGIYHCSGTGRHRYTSAGVSITVKELFTTPVLRASVSSPFPEGSLVTLNCETNLLLQRPGLQLHFSFYVGSKILEYRNTSSEYHIARAEREDAGFYWCEVATEDSSVLKRSPELELQVLGPQSSAPVWFHILFYLSVGIMFSLNTVLYVKIHRLQREKKYNLEVPLVSEQGKKANSFQQVRSDGVYEEVTATASQTTPKEAPDGPRSSVGDCGPEQPEPLPPSDSTGAQTSQS 1C4F0033842767E7 fcg2a_human Low affinity immunoglobulin gamma Fc region receptor II-a FCGR2A CD32 FCG2 FCGR2A1 IGFR2 Fc-gamma RII-a CDw32 protein protein human Homo sapiens Human MTMETQMSQNVCPRNLWLLQPLTVLLLLASADSQAAAPPKAVLKLEPPWINVLQEDSVTLTCQGARSPESDSIQWFHNGNLIPTHTQPSYRFKANNNDSGEYTCQTGQTSLSDPVHLTVLSEWLVLQTPHLEFQEGETIMLRCHSWKDKPLVKVTFFQNGKSQKFSHLDPTFSIPQANHSHSGDYHCTGNIGYTLFSSKPVTITVQVPSMGSSSPMGIIVAVVIATAVAAIVAAVVALIYCRKKRISANSTDPVKAAQFEPPGRQMIAIRKRQLEETNNDYETADGGYMTLNPRAPTDDDKNIYLTLPPNDHVNSNN 07F73F3BB282DFF6 casp8_mouse Caspase-8 Casp8 protein protein mouse Mus musculus Mouse MDFQSCLYAIAEELGSEDLAALKFLCLDYIPHKKQETIEDAQKLFLRLREKGMLEEGNLSFLKELLFHISRWDLLVNFLDCNREEMVRELRDPDNAQISPYRVMLFKLSEEVSELELRSFKFLLNNEIPKCKLEDDLSLLEIFVEMEKRTMLAENNLETLKSICDQVNKSLLGKIEDYERSSTERRMSLEGREELPPSVLDEMSLKMAELCDSPREQDSESRTSDKVYQMKNKPRGYCLIINNHDFSKAREDITQLRKMKDRKGTDCDKEALSKTFKELHFEIVSYDDCTANEIHEILEGYQSADHKNKDCFICCILSHGDKGVVYGTDGKEASIYDLTSYFTGSKCPSLSGKPKIFFIQACQGSNFQKGVPDEAGFEQQNHTLEVDSSSHKNYIPDEADFLLGMATVKNCVSYRDPVNGTWYIQSLCQSLRERCPQGDDILSILTGVNYDVSNKDDRRNKGKQMPQPTFTLRKKLFFPP DIP protein O89110 original sequence version: 82 DIP protein O89110 original sequence version: 143 DIP protein O89110 original sequence version: 151 045268AE3DE5ED4F fcg3a_human Low affinity immunoglobulin gamma Fc region receptor III-A FCGR3A CD16A FCG3 FCGR3 IGFR3 CD16a antigen Fc-gamma RIII-alpha FcR-10 IgG Fc receptor III-2 protein protein human Homo sapiens Human MWQLLLPTALLLLVSAGMRTEDLPKAVVFLEPQWYRVLEKDSVTLKCQGAYSPEDNSTQWFHNESLISSQASSYFIDAATVDDSGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRWVFKEEDPIHLRCHSWKNTALHKVTYLQNGKGRKYFHHNSDFYIPKATLKDSGSYFCRGLFGSKNVSSETVNITITQGLAVSTISSFFPPGYQVSFCLVMVLLFAVDTGLYFSVKTNIRSSTRDWKDHKFKWRKDPQDK D38D178D32C67337 tr10b_mouse Tumor necrosis factor receptor superfamily member 10B Tnfrsf10b Dr5 Killer Death receptor 5 MK protein protein mouse Mus musculus Mouse MEPPGPSTPTASAAARADHYTPGLRPLPKRRLLYSFALLLAVLQAVFVPVTANPAHNRPAGLQRPEESPSRGPCLAGQYLSEGNCKPCREGIDYTSHSNHSLDSCILCTVCKEDKVVETRCNITTNTVCRCKPGTFEDKDSPEICQSCSNCTDGEEELTSCTPRENRKCVSKTAWASWHKLGLWIGLLVPVVLLIGALLVWKTGAWRQWLLCIKRGCERDPESANSVHSSLLDRQTSSTTNDSNHNTEPGKTQKTGKKLLVPVNGNDSADDLKFIFEYCSDIVPFDSWNRLMRQLGLTDNQIQMVKAETLVTREALYQMLLKWRHQTGRSASINHLLDALEAVEERDAMEKIEDYAVKSGRFTYQNAAAQPETGPGGSQCV 222531758F4ADE0A q8r2r4-drozitumab 2 6 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association Fig 1D. fcgr1-drozitumab 2 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Fig 1D. fcgr2-drozitumab 2 4 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association Fig 1D. fcgr3-drozitumab 2 5 unspecified role unspecified role prey prey 3 unspecified role unspecified role bait bait physical association physical association Fig 1D. tr10b-casp8 1 11 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association Fig 1H. Caspase-8 recruitment was evident within 60 min of WT drozitumab treatment, with substantial processing and appearance of the p18 catalytic subunit by 180 min. Rather than just adding a comment, should this be 2 interactions - one at 60 and one at 180 mins? The p18 chain can be added as additional prey (?) at 180 mins? Cannot add the other multiple auto-catalysis cleavage products of caspase 8. Have not added Fig S1C as nothing states it is an IP. drozitumab and to a lesser extent DANA-drozitumab (mutant of IgG1 Fc region (D265A and N297A). fcgr2a-drozitumab 2 8 unspecified role unspecified role bait bait ecd_domain sufficient to bind sufficient binding region sufficient to bind range range range range c_terminal 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 c_terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role prey prey physical association physical association Fig 6A. anti-κ antibody drozitumab exhibited approximately 2- to 3-fold higher affinity for recombinant human FcγRIIA131H versus FcγRIIA131R -(natural variants/polymophisms). fcgr3a-drozitumab 2 10 unspecified role unspecified role bait bait c_terminal 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 ecd_domain sufficient to bind sufficient binding region sufficient to bind range range range range c_terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role prey prey physical association physical association Fig 6C. anti-κ antibody drozitumab exhibited higher affinity for recombinant FcγRIIIA158V versus FcγRIIIA158F- (natural variants/polymorphisms)