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/ Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell (0092-8674) 2012 imex curation Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn Accepted 2012-FEB-01 by MGT 14-Mar-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell (0092-8674) 2012 imex curation Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions human-ht_29 Homo sapiens HT-29 caucasian women with colon adenocarcinoma ht_29 HT-29 confirmed as human with IEF of AST, LDH established from the primary tumor of a 44-year-old Caucasian woman with colon adenocarcinoma in 1964; described to be heterotransplantable forming well-differentiated grade I tumors anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn Accepted 2012-FEB-01 by MGT Cell (0092-8674) 2012 Only protein-protein interactions imex curation Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell (0092-8674) 2012 imex curation Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro comigration in sds comigration in sds page comigration in sds western blot western blot Immuno blot Immuno blot Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn Accepted 2012-FEB-01 by MGT 14-Mar-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell (0092-8674) 2012 imex curation Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn Accepted 2012-FEB-01 by MGT 14-Mar-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell (0092-8674) 2012 imex curation Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn Accepted 2012-FEB-01 by MGT 14-Mar-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell (0092-8674) 2012 imex curation Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn Accepted 2012-FEB-01 by MGT 14-Mar-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Mixed Lineage Kinase Domain-like Protein Mediates Necrosis Signaling Downstream of RIP3 Kinase. Cell (0092-8674) 2012 imex curation Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip sequence tag sequence tag identification sequence tag Sun L., Wang H., Wang Z., He S., Chen S., Liao D., Wang L., Yan J., Liu W., Lei X., Wang X. Cells treated with necrosis inducers in the presence of necrosulfonamide. leixiaoguang@nibs.ac.cn, wangxiaodong@nibs.ac.cn Accepted 2012-FEB-01 by MGT 14-Mar-2012: Contacted by JYOTI. Cell (0092-8674) 2012 Only protein-protein interactions imex curation 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 DIP protein Q9Y572 original sequence version: 134 BF755F8A0B1810A1 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 DIP protein Q13546 original sequence version: 158 DIP protein Q13546-1 original sequence version: 140 DIP protein Q13546 original sequence version: 146 DIP protein Q13546-1 original sequence version: 146 DIP protein Q13546 original sequence version: 134 DIP protein Q13546 original sequence version: 109 976E2428D525A9B2 mlkl_mouse Mixed lineage kinase domain-like protein Mlkl protein protein mouse Mus musculus Mouse MDKLGQIIKLGQLIYEQCEKMKYCRKQCQRLGNRVHGLLQPLQRLQAQGKKNLPDDITAALGRFDEVLKEANQQIEKFSKKSHIWKFVSVGNDKILFHEVNEKLRDVWEELLLLLQVYHWNTVSDVSQPASWQQEDRQDAEEDGNENMKVILMQLQISVEEINKTLKQCSLKPTQEIPQDLQIKEIPKEHLGPPWTKLKTSKMSTIYRGEYHRSPVTIKVFNNPQAESVGIVRFTFNDEIKTMKKFDSPNILRIFGICIDQTVKPPEFSIVMEYCELGTLRELLDREKDLTMSVRSLLVLRAARGLYRLHHSETLHRNISSSSFLVAGGYQVKLAGFELSKTQNSISRTAKSTKAERSSSTIYVSPERLKNPFCLYDIKAEIYSFGIVLWEIATGKIPFEGCDSKKIRELVAEDKKQEPVGQDCPELLREIINECRAHEPSQRPSVDGRSLSGRERILERLSAVEESTDKKV 75E0E3E3DDB5DFB2 necrosulfonamide small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mlkl_human Mixed lineage kinase domain-like protein MLKL protein protein human Homo sapiens Human MENLKHIITLGQVIHKRCEEMKYCKKQCRRLGHRVLGLIKPLEMLQDQGKRSVPSEKLTTAMNRFKAALEEANGEIEKFSNRSNICRFLTASQDKILFKDVNRKLSDVWKELSLLLQVEQRMPVSPISQGASWAQEDQQDADEDRRAFQMLRRDNEKIEASLRRLEINMKEIKETLRQYLPPKCMQEIPQEQIKEIKKEQLSGSPWILLRENEVSTLYKGEYHRAPVAIKVFKKLQAGSIAIVRQTFNKEIKTMKKFESPNILRIFGICIDETVTPPQFSIVMEYCELGTLRELLDREKDLTLGKRMVLVLGAARGLYRLHHSEAPELHGKIRSSNFLVTQGYQVKLAGFELRKTQTSMSLGTTREKTDRVKSTAYLSPQELEDVFYQYDVKSEIYSFGIVLWEIATGDIPFQGCNSEKIRKLVAVKRQQEPLGEDCPSELREIIDECRAHDPSVRPSVDEILKKLSTFSK 005E7F7AB6D20649 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 DIP protein Q9QZL0 original sequence version: 88 DIP protein Q9QZL0 original sequence version: 125 DIP protein Q9QZL0 original sequence version: 133 31C5EEE77F55778B ripk3-ripk1-2 2 8 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey physical association physical association Fig. 1G The interaction between RIP1 and RIP3 was enhanced by Necrosulfonamide. ripk3-mlkl 7 12 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 2A ripk3-mlkl-1 2 8 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role prey prey physical association physical association Fig. 2B ripk3-mlkl-2 5 12 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey physical association physical association Fig. 2D Necrostatin-1 Necrosulfonamide MLKL knockdown enhanced RIP1/RIP3 interaction, but this interaction was not further enhanced by necrosulfonamide treatment after MLKL knockdown. necrosulfonam-mlkl-1 1 12 unspecified role unspecified role prey prey human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 11 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 8 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 association association Fig. 3C Non-biotin-labeled necrosulfonamide competed off MLKL and RIP3 binding to the beads. endogenous RIP3 was also pulled down with MLKL before necrosis induction, confirming there was a basal level of MLKL/RIP3 interaction in living cells. As necrosis progressed, less MLKL/RIP3 bound to the biotin-NSA/streptavidin-conjugated beads. necrosulfonamid-mlkl 1 12 unspecified role unspecified role prey prey n_terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range Q8NB16:p.Cys86Ser mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 20:30:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys86ser' Tue Jun 30 18:16:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys86Ser' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 11 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Figs. 3D, 4B, 4D, Supp. Figs. 4C, 5E Mouse MLKL does not bind necrosulfonamide. mlkl-necrosulfonamid 3 12 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig. 3E Non-biotin-labeled necrosulfonamide competed off MLKL and RIP3 binding to the beads. mlkl-ripk3 4 12 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9Y572:p.Ser227Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:24:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser227ala' Tue Jun 30 16:54:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser227Ala' Q9Y572:p.Asp142Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D N Wed May 02 15:52:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp142asn' Tue Jun 30 18:18:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp142Asn' region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 5B, 7C. ripk3-mlkl-4 4 8 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 61 65 physical association physical association Fig. 5C. mlkl-ripk3-1 6 12 enzyme target enzyme target substrate substrate neutral component neutral component Q8NB16:p.Thr357_Ser358delinsAlaAla mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain TS AA Thu Jul 07 12:07:40 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was thr357_ser358ala Fri Jul 22 11:14:53 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'thr357ala,ser358ala' Wed May 02 15:48:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr_ser357-358ala_ala' Tue Jun 30 17:53:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr357_Ser358delinsAlaAla' Q8NB16:p.Thr357_Ser358delinsGluAsp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain TS ED Thu Jul 07 12:07:41 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was thr357glu_ser358asp Fri Jul 22 11:08:47 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'thr357glu,ser358asp' Wed May 02 15:59:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr_ser357-358glu_asp' Tue Jun 30 18:30:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr357_Ser358delinsGluAsp' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 enzyme enzyme neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 physical association physical association Fig. 5E, 6A, 6C, Supp. Fig. 5A Enzyme produced by coip but gel in Fig. 2A shows this to be very clean so have added kinase assay. Tandem mass spectrometry of MLKL unambiguously identified threonine 357 and serine 358 as being phosphorylated necrosulfonam-mlkl-2 1 11 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 12 unspecified role unspecified role prey prey human-ht_29 Homo sapiens HT-29 caucasian women with colon adenocarcinoma ht_29 HT-29 confirmed as human with IEF of AST, LDH established from the primary tumor of a 44-year-old Caucasian woman with colon adenocarcinoma in 1964; described to be heterotransplantable forming well-differentiated grade I tumors 8 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-ht_29 Homo sapiens HT-29 caucasian women with colon adenocarcinoma ht_29 HT-29 confirmed as human with IEF of AST, LDH established from the primary tumor of a 44-year-old Caucasian woman with colon adenocarcinoma in 1964; described to be heterotransplantable forming well-differentiated grade I tumors association association Supp. Fig. 3B Non-biotin-labeled necrosulfonamide competed off MLKL and RIP3 binding to the beads. endogenous RIP3 was also pulled down with MLKL before necrosis induction, confirming there was a basal level of MLKL/RIP3 interaction in living cells. As necrosis progressed, less MLKL/RIP3 bound to the biotin-NSA/streptavidin-conjugated beads. ripk3-mlkl-9 5 8 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9Y572:p.Lys50Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K A Wed May 02 14:41:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys50ala' Tue Jun 30 20:17:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys50Ala' 12 unspecified role unspecified role prey prey physical association physical association Fig. 5D ripk3-mlkl-10 4 8 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. S5D mlkl-ripk3-2 4 10 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association S6 Mouse RIP3 only binds to mouse MLKL, but not the human homolog.