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/ cho-2009-1 Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. human-jurkat Homo sapiens jurkat Human T cell leukemia anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cho YS., Challa S., Moquin D., Genga R., Ray TD., Guildford M., Chan FK. Apoptosis - Interactions involving proteins with a function related to apoptosis francis.chan@umassmed.edu Cell (0092-8674) 2009 Only protein-protein interactions 16 imex curation cho-2009-2 Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Cho YS., Challa S., Moquin D., Genga R., Ray TD., Guildford M., Chan FK. Apoptosis - Interactions involving proteins with a function related to apoptosis francis.chan@umassmed.edu Cell (0092-8674) 2009 Only protein-protein interactions 16 imex curation cho-2009-4 Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cho YS., Challa S., Moquin D., Genga R., Ray TD., Guildford M., Chan FK. Apoptosis - Interactions involving proteins with a function related to apoptosis francis.chan@umassmed.edu Cell (0092-8674) 2009 Only protein-protein interactions 16 imex curation cho-2009-6 Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Cho YS., Challa S., Moquin D., Genga R., Ray TD., Guildford M., Chan FK. Apoptosis - Interactions involving proteins with a function related to apoptosis Additional information on constructs obtained from the author. francis.chan@umassmed.edu Cell (0092-8674) 2009 Only protein-protein interactions 16 imex curation cho-2009-3 Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. human-jurkat Homo sapiens jurkat Human T cell leukemia anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cho YS., Challa S., Moquin D., Genga R., Ray TD., Guildford M., Chan FK. Apoptosis - Interactions involving proteins with a function related to apoptosis francis.chan@umassmed.edu Cell (0092-8674) 2009 Only protein-protein interactions 16 imex curation cho-2009-5 Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. mouse-liver Mus musculus liver liver anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Cho YS., Challa S., Moquin D., Genga R., Ray TD., Guildford M., Chan FK. Apoptosis - Interactions involving proteins with a function related to apoptosis francis.chan@umassmed.edu Cell (0092-8674) 2009 Only protein-protein interactions 16 imex curation cho-2009-7 Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Cho YS., Challa S., Moquin D., Genga R., Ray TD., Guildford M., Chan FK. Apoptosis - Interactions involving proteins with a function related to apoptosis Additional information on constructs obtained from the author. francis.chan@umassmed.edu Cell (0092-8674) 2009 Only protein-protein interactions 16 imex curation ripk1_human Receptor-interacting serine/threonine-protein kinase 1 RIPK1 RIP Cell death protein RIP RIP1 Receptor-interacting protein 1 protein protein human Homo sapiens Human MQPDMSLNVIKMKSSDFLESAELDSGGFGKVSLCFHRTQGLMIMKTVYKGPNCIEHNEALLEEAKMMNRLRHSRVVKLLGVIIEEGKYSLVMEYMEKGNLMHVLKAEMSTPLSVKGRIILEIIEGMCYLHGKGVIHKDLKPENILVDNDFHIKIADLGLASFKMWSKLNNEEHNELREVDGTAKKNGGTLYYMAPEHLNDVNAKPTEKSDVYSFAVVLWAIFANKEPYENAICEQQLIMCIKSGNRPDVDDITEYCPREIISLMKLCWEANPEARPTFPGIEEKFRPFYLSQLEESVEEDVKSLKKEYSNENAVVKRMQSLQLDCVAVPSSRSNSATEQPGSLHSSQGLGMGPVEESWFAPSLEHPQEENEPSLQSKLQDEANYHLYGSRMDRQTKQQPRQNVAYNREEERRRRVSHDPFAQQRPYENFQNTEGKGTAYSSAASHGNAVHQPSGLTSQPQVLYQNNGLYSSHGFGTRPLDPGTAGPRVWYRPIPSHMPSLHNIPVPETNYLGNTPTMPFSSLPPTDESIKYTIYNSTGIQIGAYNYMEIGGTSSSLLDSTNTNFKEEPAAKYQAIFDNTTSLTDKHLDPIRENLGKHWKNCARKLGFTQSQIDEIDHDYERDGLKEKVYQMLQKWVMREGIKGATVGKLAQALHQCSRIDLLSSLIYVSQN 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 ripk3_human Receptor-interacting serine/threonine-protein kinase 3 RIPK3 RIP3 RIP-like protein kinase 3 Receptor-interacting protein 3 protein protein human Homo sapiens Human MSCVKLWPSGAPAPLVSIEELENQELVGKGGFGTVFRAQHRKWGYDVAVKIVNSKAISREVKAMASLDNEFVLRLEGVIEKVNWDQDPKPALVTKFMENGSLSGLLQSQCPRPWPLLCRLLKEVVLGMFYLHDQNPVLLHRDLKPSNVLLDPELHVKLADFGLSTFQGGSQSGTGSGEPGGTLGYLAPELFVNVNRKASTASDVYSFGILMWAVLAGREVELPTEPSLVYEAVCNRQNRPSLAELPQAGPETPGLEGLKELMQLCWSSEPKDRPSFQECLPKTDEVFQMVENNMNAAVSTVKDFLSQLRSSNRRFSIPESGQGGTEMDGFRRTIENQHSRNDVMVSEWLNKLNLEEPPSSVPKKCPSLTKRSRAQEEQVPQAWTAGTSSDSMAQPPQTPETSTFRNQMPSPTSTGTPSPGPRGNQGAERQGMNWSCRTPEPNPVTGRPLVNIYNCSGVQVGDNNYLTMQQTTALPTWGLAPSGKGRGLQHPPPVGSQEGPKDPEAWSRPQGWYNHSGK tnr1a_human Tumor necrosis factor receptor superfamily member 1A TNFRSF1A TNFAR TNFR1 p60 Tumor necrosis factor receptor 1 Tumor necrosis factor receptor type I p55 protein protein human Homo sapiens Human MGLSTVPDLLLPLVLLELLVGIYPSGVIGLVPHLGDREKRDSVCPQGKYIHPQNNSICCTKCHKGTYLYNDCPGPGQDTDCRECESGSFTASENHLRHCLSCSKCRKEMGQVEISSCTVDRDTVCGCRKNQYRHYWSENLFQCFNCSLCLNGTVHLSCQEKQNTVCTCHAGFFLRENECVSCSNCKKSLECTKLCLPQIENVKGTEDSGTTVLLPLVIFFGLCLLSLLFIGLMYRYQRWKSKLYSIVCGKSTPEKEGELEGTTTKPLAPNPSFSPTPGFTPTLGFSPVPSSTFTSSSTYTPGDCPNFAAPRREVAPPYQGADPILATALASDPIPNPLQKWEDSAHKPQSLDTDDPATLYAVVENVPPLRWKEFVRRLGLSDHEIDRLELQNGRCLREAQYSMLATWRRRTPRREATLELLGRVLRDMDLLGCLEDIEEALCGPAALPPAPSLLR 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 tradd_human Tumor necrosis factor receptor type 1-associated DEATH domain protein TRADD TNFRSF1A-associated via death domain protein protein human Homo sapiens Human MAAGQNGHEEWVGSAYLFVESSLDKVVLSDAYAHPQQKVAVYRALQAALAESGGSPDVLQMLKIHRSDPQLIVQLRFCGRQPCGRFLRAYREGALRAALQRSLAAALAQHSVPLQLELRAGAERLDALLADEERCLSCILAQQPDRLRDEELAELEDALRNLKCGSGARGGDGEVASAPLQPPVPSLSEVKPPPPPPPAQTFLFQGQPVVNRPLSLKDQQTFARSVGLKWRKVGRSLQRGCRALRDPALDSLAYEYEREGLYEQAFQLLRRFVQAEGRRATLQRLVEALEENELTSLAEDLLGLTDPNGGLA ripk1_mouse Receptor-interacting serine/threonine-protein kinase 1 Cell death protein RIP Receptor-interacting protein 1 Ripk1 Rinp Rip protein protein mouse Mus musculus Mouse MQPDMSLDNIKMASSDLLEKTDLDSGGFGKVSLCYHRSHGFVILKKVYTGPNRAEYNEVLLEEGKMMHRLRHSRVVKLLGIIIEEGNYSLVMEYMEKGNLMHVLKTQIDVPLSLKGRIIVEAIEGMCYLHDKGVIHKDLKPENILVDRDFHIKIADLGVASFKTWSKLTKEKDNKQKEVSSTTKKNNGGTLYYMAPEHLNDINAKPTEKSDVYSFGIVLWAIFAKKEPYENVICTEQFVICIKSGNRPNVEEILEYCPREIISLMERCWQAIPEDRPTFLGIEEEFRPFYLSHFEEYVEEDVASLKKEYPDQSPVLQRMFSLQHDCVPLPPSRSNSEQPGSLHSSQGLQMGPVEESWFSSSPEYPQDENDRSVQAKLQEEASYHAFGIFAEKQTKPQPRQNEAYNREEERKRRVSHDPFAQQRARENIKSAGARGHSDPSTTSRGIAVQQLSWPATQTVWNNGLYNQHGFGTTGTGVWYPPNLSQMYSTYKTPVPETNIPGSTPTMPYFSGPVADDLIKYTIFNSSGIQIGNHNYMDVGLNSQPPNNTCKEESTSRHQAIFDNTTSLTDEHLNPIRENLGRQWKNCARKLGFTESQIDEIDHDYERDGLKEKVYQMLQKWLMREGTKGATVGKLAQALHQCCRIDLLNHLIRASQS fadd_mouse FAS-associated death domain protein FAS-associating death domain-containing protein Mediator of receptor induced toxicity Fadd Mort1 protein protein mouse Mus musculus Mouse MDPFLVLLHSLSGSLSGNDLMELKFLCRERVSKRKLERVQSGLDLFTVLLEQNDLERGHTGLLRELLASLRRHDLLQRLDDFEAGTATAAPPGEADLQVAFDIVCDNVGRDWKRLARELKVSEAKMDGIEEKYPRSLSERVRESLKVWKNAEKKNASVAGLVKALRTCRLNLVADLVEEAQESVSKSENMSPVLRDSTVSSSETP ripk3_mouse Receptor-interacting serine/threonine-protein kinase 3 Ripk3 Rip3 RIP-like protein kinase 3 Receptor-interacting protein 3 protein protein mouse Mus musculus Mouse MSSVKLWPTGASAVPLVSREELKKLEFVGKGGFGVVFRAHHRTWNHDVAVKIVNSKKISWEVKAMVNLRNENVLLLLGVTEDLQWDFVSGQALVTRFMENGSLAGLLQPECPRPWPLLCRLLQEVVLGMCYLHSLNPPLLHRDLKPSNILLDPELHAKLADFGLSTFQGGSQSGSGSGSGSRDSGGTLAYLDPELLFDVNLKASKASDVYSFGILVWAVLAGREAELVDKTSLIRETVCDRQSRPPLTELPPGSPETPGLEKLKELMIHCWGSQSENRPSFQDCEPKTNEVYNLVKDKVDAAVSEVKHYLSQHRSSGRNLSAREPSQRGTEMDCPRETMVSKMLDRLHLEEPSGPVPGKCPERQAQDTSVGPATPARTSSDPVAGTPQIPHTLPFRGTTPGPVFTETPGPHPQRNQGDGRHGTPWYPWTPPNPMTGPPALVFNNCSEVQIGNYNSLVAPPRTTASSSAKYDQAQFGRGRGWQPFHK casp8-ripk1-2 Interaction between CASP8, RIPK1 and RIPK3 834166 n/a 834173 unspecified role unspecified role prey prey n/a 834174 unspecified role unspecified role bait bait n/a 834175 unspecified role unspecified role prey prey association association false false false TNF-alpha plus zVAD-fmk (necrosis inducer) Fig. 2E tnfrsf1a-ripk1-1 Interaction between TNFSF1A and RIPK1 demonstrated by coimmunoprecipitation 834166 n/a 834173 unspecified role unspecified role prey prey n/a 834176 unspecified role unspecified role bait bait physical association physical association false false false TNF-alpha plus zVAD-fmk (necrosis inducer) Fig. 2F fadd-ripk1-1 Interaction between FADD, RIPK1 and RIPK3 834166 n/a 834177 unspecified role unspecified role bait bait n/a 834175 unspecified role unspecified role prey prey n/a 834173 unspecified role unspecified role prey prey association association false false false Figs. 3A, 3D In wild-type cells, complex had kinase activity, however TNFR2- cells that do not undergo programmed necrosis in response to TNF, also exhibited no inducible FADD complex II kinase activity. trnadd-ripk1-2 Interaction between TRADD, RIPK1 and TNFRSF1A 834166 n/a 834176 unspecified role unspecified role bait bait n/a 834173 unspecified role unspecified role prey prey n/a 834178 unspecified role unspecified role prey prey association association false false false Fig. 3C Complex lacked kinase activity casp8-ripk1-3 Interaction between CASP8, RIPK1 and TRADD 834166 n/a 834178 unspecified role unspecified role prey prey n/a 834173 unspecified role unspecified role prey prey n/a 834174 unspecified role unspecified role bait bait association association false false false TNF-alpha plus zVAD-fmk (necrosis inducer) RIP1 is cleaved within caspase-8-associated Complex II Supp. Fig. 3 ripk3-1 Autophosphorylation of RIPK3 834167 n/a 834175 putative self putative self putative self putative self human-jurkat Homo sapiens jurkat Human T cell leukemia phosphorylation phosphorylation reaction phosphorylation false true false Autophosphorylation of RIPK1. Fig. 3B ripk1-1 Autophosphorylation of Ripk1 834167 n/a 834179 putative self putative self putative self putative self mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. phosphorylation phosphorylation reaction phosphorylation false true false Fig. 4B Autophosphorylation of RIPK1. Autophosphorylation of RIPK1. ripk3-ripk1-1 phosphorylation of PIPK1 by RIPK3 834167 n/a 834175 enzyme enzyme neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q9Y572:p.Asp160Asn Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. n/a 834173 enzyme target enzyme target substrate substrate neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. phosphorylation phosphorylation reaction phosphorylation false false false Phosphorylation of RIPK1. Fig. 4D fadd-ripk1-3 Interaction between FADD, RIPK1 and RIPK3 demonstrated by coimmunoprecipitation 834168 n/a 834180 unspecified role unspecified role bait bait n/a 834179 unspecified role unspecified role prey prey n/a 834181 unspecified role unspecified role prey prey association association false false false Fig. 4A Interaction does not take place in Ripk3 -/- cells. ripk1-ripk3-6 Interaction between RIPK1 and RIPK3 demonstrated by coimmunoprecipitation 834168 n/a 834179 unspecified role unspecified role bait bait n/a 834181 unspecified role unspecified role prey prey physical association physical association false false false Fig. 4E, 4F TNF-alpha plus cycloheximide plus zVAD-fmk Necrosin-1 ripk1-ripk3-7 Interaction between RIPK1 and RIPK3 demonstrated by coimmunoprecipitation 834168 n/a 834181 unspecified role unspecified role bait bait n/a 834179 unspecified role unspecified role prey prey physical association physical association false false false Figs. 4G, 6F TNF-alpha plus cycloheximide plus zVAD-fmk. TNF-alpha laone is enough to trigger the interaction in virally (vaccinia virus) infected cells. Necrosin-1 ripk1-ripk3-9 Interaction between RIPK1 and RIPK3 demonstrated by coimmunoprecipitation 834169 n/a 834175 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 834173 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rhim region required to bind necessary binding region required to bind mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Supp. Fig. 2 ripk1-ripk3-10 Interaction between RIPK1 and RIPK3 demonstrated by coimmunoprecipitation 834169 n/a 834175 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 834173 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false rhim region required to bind necessary binding region required to bind mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Supp. Fig. 2 fadd-ripk1-2 Interaction between FADD and RIPK3 834170 n/a 834175 unspecified role unspecified role prey prey n/a 834177 unspecified role unspecified role bait bait association association false false false Figs. 3E In wild-type cells, complex had kinase activity, however TNFR2- cells that do not undergo programmed necrosis in response to TNF, also exhibited no inducible FADD complex II kinase activity. ripk1-ripk3-8 Interaction between RIPK1 and RIPK3 demonstrated by coimmunoprecipitation 834171 n/a 834181 unspecified role unspecified role prey prey n/a 834179 unspecified role unspecified role bait bait physical association physical association false false false Fig. 7E Vaccinia virus infection ripk1-fadd-1 Interaction between RIPK1 and FADD demonstrated by coimmunoprecipitation 834172 n/a 834177 unspecified role unspecified role prey prey n/a 834173 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. Fig. 4C