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/ wilson-2011-1 An Fcγ receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. 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 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 Cancer Cell (1535-6108) 2011 Only protein-protein interactions imex curation wilson-2011-2 An Fcγ receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. 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 Cancer Cell (1535-6108) 2011 Only protein-protein interactions imex curation fcgr4_mouse Low affinity immunoglobulin gamma Fc region receptor IV Fcgr4 CD16-2 FcgammaRIV protein protein mouse Mus musculus Mouse MWQLLLPTALVLTAFSGIQAGLQKAVVNLDPKWVRVLEEDSVTLRCQGTFSPEDNSIKWFHNESLIPHQDANYVIQSARVKDSGMYRCQTALSTISDPVQLEVHMGWLLLQTTKWLFQEGDPIHLRCHSWQNRPVRKVTYLQNGKGKKYFHENSELLIPKATHNDSGSYFCRGLIGHNNKSSASFRISLGDPGSPSMFPPWHQITFCLLIGLLFAIDTVLYFSVRRGLQSPVADYEEPKIQWSKEPQDK drozitumab drozitumab-anti-DR5(CD262)Human IgG1 heavy chain, disulfide linked to human λ3-chain, dimer. protein protein human Homo sapiens Human fcgr1_mouse High affinity immunoglobulin gamma Fc receptor I Fcgr1 Fcg1 Fc-gamma RI protein protein mouse Mus musculus Mouse MILTSFGDDMWLLTTLLLWVPVGGEVVNATKAVITLQPPWVSIFQKENVTLWCEGPHLPGDSSTQWFINGTAVQISTPSYSIPEASFQDSGEYRCQIGSSMPSDPVQLQIHNDWLLLQASRRVLTEGEPLALRCHGWKNKLVYNVVFYRNGKSFQFSSDSEVAILKTNLSHSGIYHCSGTGRHRYTSAGVSITVKELFTTPVLRASVSSPFPEGSLVTLNCETNLLLQRPGLQLHFSFYVGSKILEYRNTSSEYHIARAEREDAGFYWCEVATEDSSVLKRSPELELQVLGPQSSAPVWFHILFYLSVGIMFSLNTVLYVKIHRLQREKKYNLEVPLVSEQGKKANSFQQVRSDGVYEEVTATASQTTPKEAPDGPRSSVGDCGPEQPEPLPPSDSTGAQTSQS 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 fcgr3_mouse Low affinity immunoglobulin gamma Fc region receptor III Fcgr3 Fc-gamma RIII protein protein mouse Mus musculus Mouse MFQNAHSGSQWLLPPLTILLLFAFADRQSAALPKAVVKLDPPWIQVLKEDMVTLMCEGTHNPGNSSTQWFHNGRSIRSQVQASYTFKATVNDSGEYRCQMEQTRLSDPVDLGVISDWLLLQTPQRVFLEGETITLRCHSWRNKLLNRISFFHNEKSVRYHHYKSNFSIPKANHSHSGDYYCKGSLGSTQHQSKPVTITVQDPATTSSISLVWYHTAFSLVMCLLFAVDTGLYFYVRRNLQTPREYWRKSLSIRKHQAPQDK 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 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 tr10b_mouse Tumor necrosis factor receptor superfamily member 10B Tnfrsf10b Dr5 Killer Death receptor 5 MK protein protein mouse Mus musculus Mouse MEPPGPSTPTASAAARADHYTPGLRPLPKRRLLYSFALLLAVLQAVFVPVTANPAHNRPAGLQRPEESPSRGPCLAGQYLSEGNCKPCREGIDYTSHSNHSLDSCILCTVCKEDKVVETRCNITTNTVCRCKPGTFEDKDSPEICQSCSNCTDGEEELTSCTPRENRKCVSKTAWASWHKLGLWIGLLVPVVLLIGALLVWKTGAWRQWLLCIKRGCERDPESANSVHSSLLDRQTSSTTNDSNHNTEPGKTQKTGKKLLVPVNGNDSADDLKFIFEYCSDIVPFDSWNRLMRQLGLTDNQIQMVKAETLVTREALYQMLLKWRHQTGRSASINHLLDALEAVEERDAMEKIEDYAVKSGRFTYQNAAAQPETGPGGSQCV casp8_mouse Caspase-8 Casp8 protein protein mouse Mus musculus Mouse MDFQSCLYAIAEELGSEDLAALKFLCLDYIPHKKQETIEDAQKLFLRLREKGMLEEGNLSFLKELLFHISRWDLLVNFLDCNREEMVRELRDPDNAQISPYRVMLFKLSEEVSELELRSFKFLLNNEIPKCKLEDDLSLLEIFVEMEKRTMLAENNLETLKSICDQVNKSLLGKIEDYERSSTERRMSLEGREELPPSVLDEMSLKMAELCDSPREQDSESRTSDKVYQMKNKPRGYCLIINNHDFSKAREDITQLRKMKDRKGTDCDKEALSKTFKELHFEIVSYDDCTANEIHEILEGYQSADHKNKDCFICCILSHGDKGVVYGTDGKEASIYDLTSYFTGSKCPSLSGKPKIFFIQACQGSNFQKGVPDEAGFEQQNHTLEVDSSSHKNYIPDEADFLLGMATVKNCVSYRDPVNGTWYIQSLCQSLRERCPQGDDILSILTGVNYDVSNKDDRRNKGKQMPQPTFTLRKKLFFPP q8r2r4-drozitumab 1017911 n/a 1017913 unspecified role unspecified role bait bait n/a 1017914 unspecified role unspecified role prey prey physical association physical association false false false Fig 1D. fcgr1-drozitumab 1017911 n/a 1017914 unspecified role unspecified role prey prey n/a 1017915 unspecified role unspecified role bait bait physical association physical association false false false Fig 1D. fcgr2-drozitumab 1017911 n/a 1017916 unspecified role unspecified role bait bait n/a 1017914 unspecified role unspecified role prey prey physical association physical association false false false Fig 1D. fcgr3-drozitumab 1017911 n/a 1017914 unspecified role unspecified role prey prey n/a 1017917 unspecified role unspecified role bait bait physical association physical association false false false Fig 1D. fcgr2a-drozitumab 1017911 n/a 1017918 unspecified role unspecified role bait bait ecd_domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false 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 false n/a 1017914 unspecified role unspecified role prey prey physical association physical association false false false 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 1017911 n/a 1017919 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 false ecd_domain sufficient to bind sufficient binding region sufficient to bind range range range range false 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 false n/a 1017914 unspecified role unspecified role prey prey physical association physical association false false false Fig 6C. anti-κ antibody drozitumab exhibited higher affinity for recombinant FcγRIIIA158V versus FcγRIIIA158F- (natural variants/polymorphisms) tr10b-casp8 1017912 n/a 1017920 unspecified role unspecified role bait bait n/a 1017921 unspecified role unspecified role prey prey physical association physical association false false false 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. drozitumab and to a lesser extent DANA-drozitumab (mutant of IgG1 Fc region (D265A and N297A).