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/ Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Blood 2013 imex curation Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting tag visualisation tag visualisation Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr Accepted 2017-JUN-14 AT 09:16 BST AT 09:16 BST by ORCHARD Blood 2013 imex curation These cannot be standard CoIPs as a primary antibody was NOT used for detection; they have just added NKp44-Ig fusion protein directly to membranes and detected if it has bound using an HRP coupled secondary antibody to Ig (method section is not specific here). In Far Western "probe" is the Bait (NKp44-Ig fusion protein). (if you do not like "Far Western" as a method - maybe it could be a solid phase assay?) Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Blood 2013 imex curation Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr 14-Jul-2017: Contacted by IntAct-Help. 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 fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr Accepted 2017-JUN-14 AT 09:16 BST AT 09:16 BST by ORCHARD Blood 2013 imex curation These cannot be standard CoIPs as a primary antibody was NOT used for detection; they have just added NKp44-Ig fusion protein directly to membranes and detected if it has bound using an HRP coupled secondary antibody to Ig (method section is not specific here). In Far Western "probe" is the Bait (NKp44-Ig fusion protein). (if you do not like "Far Western" as a method - maybe it could be a solid phase assay?) Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Blood 2013 imex curation Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions human-wm1361 human-wm1361 melanoma cell line Human wm1361 wm1361 melanoma cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr Accepted 2017-JUN-14 AT 09:16 BST AT 09:16 BST by ORCHARD Blood 2013 imex curation These cannot be standard CoIPs as a primary antibody was NOT used for detection; they have just added NKp44-Ig fusion protein directly to membranes and detected if it has bound using an HRP coupled secondary antibody to Ig (method section is not specific here). In Far Western "probe" is the Bait (NKp44-Ig fusion protein). (if you do not like "Far Western" as a method - maybe it could be a solid phase assay?) Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Identification of a cellular ligand for the natural cytotoxicity receptor NKp44. Blood 2013 imex curation Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr 14-Jul-2017: Contacted by IntAct-Help. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast lexa b52 complement lexa b52 complementation LexA B52 transcription complementation lexa b52 complement nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Baychelier F., Sennepin A., Ermonval M., Dorgham K., Debré P., Vieillard V. NKp44L =MLL5 isoform Q8I2D2-8 vincent.vieillard@upmc.fr Accepted 2017-JUN-14 AT 09:16 BST AT 09:16 BST by ORCHARD pYESTrp Jurkat cDNA library Blood 2013 imex curation These cannot be standard CoIPs as a primary antibody was NOT used for detection; they have just added NKp44-Ig fusion protein directly to membranes and detected if it has bound using an HRP coupled secondary antibody to Ig (method section is not specific here). In Far Western "probe" is the Bait (NKp44-Ig fusion protein). (if you do not like "Far Western" as a method - maybe it could be a solid phase assay?) nctr2_human Natural cytotoxicity triggering receptor 2 NCR2 Lymphocyte antigen 95 homolog NK cell-activating receptor Natural killer cell p44-related protein LY95 protein protein human Homo sapiens Human MAWRALHPLLLLLLLFPGSQAQSKAQVLQSVAGQTLTVRCQYPPTGSLYEKKGWCKEASALVCIRLVTSSKPRTMAWTSRFTIWDDPDAGFFTVTMTDLREEDSGHYWCRIYRPSDNSVSKSVRFYLVVSPASASTQTSWTPRDLVSSQTQTQSCVPPTAGARQAPESPSTIPVPSQPQNSTLRPGPAAPIALVPVFCGLLVAKSLVLSALLVWWGDIWWKTMMELRSLDTQKATCHLQQVTDLPWTSVSSPVEREILYHTVARTKISDDDDEHTL 4B7AF3F451CA9F9E q8izd2-8 Inactive histone-lysine N-methyltransferase 2E Myeloid/lymphoid or mixed-lineage leukemia protein 5 KMT2E MLL5 protein protein human Homo sapiens Human MSIVIPLGVDTAETSYLEMAAGSEPESVEASPVVVEKSNSYPHQLYTSSSHHSHSYIGLPYADHNYGARPPPTPPASPPPSVLISKNEVGIFTTPNFDETSSATTISTSEDGSYGTDVTRCICGFTHDDGYMICCDKCSVWQHIDCMGIDRQHIPDTYLCERCQPRNLDKERAVLLQRRKRENMSDGDTSATESGDEVPVELYTAFQHTPTSITLTASRVSKVNDKRRKKSGEKEQHISKCKKAFREGSRKSSRVKGSAPEIDPSSDGSNFGWETKIKAWMDRYEEANNNQYSEGVQREAQRIALRLGNGNDKKEMNKSDLNTNNLLFKPPVESHIQKNKKILKSAKDLPPDALIIEYRGKFMLREQFEANGYFFKRPYPFVLFYSKFHGLEMCVDARTFGNEARFIRRSCTPNAEVRHEIQDGTIHLYIYSIHSIPKGTEITIAFDFDYGNCKYKVDCACLKENPECPVLKRSSESMENINSGYETRRKKGKKDKDISKEKDTQNQNITLDCEGTTNKMKSPETKQRKLSPLRLSVSNNQEPDFIDDIEEKTPISNEVEMESEEQIAERKRKMTREERKMEAILQAFARLEKREKRREQALERISTAKTEVKTECKDTQIVSDAEVIQEQAKEENASKPTPAKVNRTKQRKSFSRSRTHIGQQRRRHRTVSMCSDIQPSSPDIEVTSQQNDIENTVLTIEPETETALAEIITETEVPALNKCPTKYPKTKKHLVNEWLSEKNEKTGKPSDGLSERPLRITTDPEVLATQLNSLPGLTYSPHVYSTPKHYIRFTSPFLSEKRRRKEPTENISGSCKKRWLKQALEEENSAILHRFNSPCQERSRSPAVNGENKSPLLLNDSCSLPDLTTPLKKRRFYQLLDSVYSETSTPTPSPYATPTHTDITPMDPSFATPPRIKSDDETCRNGYKPIYSPVTPVTPGTPGNTMHFENISSPESSPEIKRRTYSQEGYDRSSTMLTLGPFRNSNLTELGLQEIKTIGYTSPRSRTEVNRQCPGEKEPVSDLQLGLDAVEPTALHKTLETPAHDRAEPNSQLDSTHSGRGTMYSSWVKSPDRTGVNFSVNSNLRDLTPSHQLEVGGGFRISESKCLMQDDTRGMFMETTVFCTSEDGLVSGFGRTVNDNLIDGNCTPQNPPQKKKSPVGNFVGSNVV B5D59D07B6ED13FA kmt2e_human Inactive histone-lysine N-methyltransferase 2E KMT2E MLL5 Myeloid/lymphoid or mixed-lineage leukemia protein 5 protein protein human Homo sapiens Human MSIVIPLGVDTAETSYLEMAAGSEPESVEASPVVVEKSNSYPHQLYTSSSHHSHSYIGLPYADHNYGARPPPTPPASPPPSVLISKNEVGIFTTPNFDETSSATTISTSEDGSYGTDVTRCICGFTHDDGYMICCDKCSVWQHIDCMGIDRQHIPDTYLCERCQPRNLDKERAVLLQRRKRENMSDGDTSATESGDEVPVELYTAFQHTPTSITLTASRVSKVNDKRRKKSGEKEQHISKCKKAFREGSRKSSRVKGSAPEIDPSSDGSNFGWETKIKAWMDRYEEANNNQYSEGVQREAQRIALRLGNGNDKKEMNKSDLNTNNLLFKPPVESHIQKNKKILKSAKDLPPDALIIEYRGKFMLREQFEANGYFFKRPYPFVLFYSKFHGLEMCVDARTFGNEARFIRRSCTPNAEVRHEIQDGTIHLYIYSIHSIPKGTEITIAFDFDYGNCKYKVDCACLKENPECPVLKRSSESMENINSGYETRRKKGKKDKDISKEKDTQNQNITLDCEGTTNKMKSPETKQRKLSPLRLSVSNNQEPDFIDDIEEKTPISNEVEMESEEQIAERKRKMTREERKMEAILQAFARLEKREKRREQALERISTAKTEVKTECKDTQIVSDAEVIQEQAKEENASKPTPAKVNRTKQRKSFSRSRTHIGQQRRRHRTVSMCSDIQPSSPDIEVTSQQNDIENTVLTIEPETETALAEIITETEVPALNKCPTKYPKTKKHLVNEWLSEKNEKTGKPSDGLSERPLRITTDPEVLATQLNSLPGLTYSPHVYSTPKHYIRFTSPFLSEKRRRKEPTENISGSCKKRWLKQALEEENSAILHRFNSPCQERSRSPAVNGENKSPLLLNDSCSLPDLTTPLKKRRFYQLLDSVYSETSTPTPSPYATPTHTDITPMDPSFATPPRIKSDDETCRNGYKPIYSPVTPVTPGTPGNTMHFENISSPESSPEIKRRTYSQEGYDRSSTMLTLGPFRNSNLTELGLQEIKTIGYTSPRSRTEVNRQCPGEKEPVSDLQLGLDAVEPTALHKTLETPAHDRAEPNSQLDSTHSGRGTMYSSWVKSPDRTGVNFSVNSNLRDLTPSHQLEVGGGFRISESKCLMQDDTRGMFMETTVFCTSEDGLVSGFGRTVNDNLIDGNCTPQNPPQKKKVSLLEYRKRQREARKSGSKTENFPLISVSPHASGSLSNNGDGCASSNDNGEQVDHTASLPLPTPATVYNATSEETSNNCPVKDATASEKNEPEVQWTASTSVEQVRERSYQRALLLSDHRKDKDSGGESPCVSCSPSHVQSSPSSHSNHIPQLQAKGPVPSFSELMEDPDPENPEPTTTNECPSPDTSQNTCKSPPKMSKPGSPGSVIPAQAHGKIFTKPDPQWDSTVSASEAENGVHLKTELQQKQLSNNNQALSKNHPPQTHVRNSSEQLSQKLPSVPTKLHCPPSPHLENPPKSSTPHTPVQHGYLSPKPPSQQLGSPYRPHHSQSPQVGTPQREPQRNFYPAAQNLPANTQQATSGTLFTQTPSGQSSATYSQFNQQSLNSTAPPPPPPPPPSSSYYQNQQPSANFQNYNQLKGSLSQQTVFTSGPNQALPGTTSQQTVPGHHVTPGHFLPSQNPTIHHQTAAAVVPPPPPPPPAPGPHLVQQPNSHQQHSVAHVVGPVHAVTPGSHIHSQTAGHHLPPPPPPPGPAPHHHPPPHPSTGLQGLQAQHQHVVNSAPPPPPPPPPSSVLASGHHTTSAQALHHPPHQGPPLFPSSAHPTVPPYPSQATHHTTLGPGPQHQPSGTGPHCPLPVTGPHLQPQGPNSIPTPTASGFCPHPGSVALPHGVQGPQQASPVPGQIPIHRAQVPPTFQNNYHGSGWH 8ACCB3CDB5BFCEFA ncr2-kmt2e-5 4 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait ecd sufficient to bind sufficient binding region sufficient to bind range range range range physical association physical association Supp Table 1 ncr2-kmt2e-1 3 6 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc physical association physical association Fig 2C ncr2-kmt2e-2 1 6 unspecified role unspecified role prey prey human-wm1361 human-wm1361 melanoma cell line Human wm1361 wm1361 melanoma cell line 5 unspecified role unspecified role bait bait ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc physical association physical association Fig 2B ncr2-kmt2e-3 1 7 unspecified role unspecified role prey prey human-wm1361 human-wm1361 melanoma cell line Human wm1361 wm1361 melanoma cell line 5 unspecified role unspecified role bait bait ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range mouse-nso NSO-mouse myeloma subclone of NS-1 ns0 NS0 Mouse Myeloma subclone of NS-1 http://www.hpacultures.org.uk/products/celllines/generalcell/detail.jsp?refId=85110503&collection=ecacc_gc from ecacc physical association physical association Fig 2D The antibody to MLL5 used was targetted to the N-terminal domain, common to all MLL5 isoforms. ncr2-kmt2e-4 2 5 unspecified role unspecified role neutral component neutral component ecd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role neutral component neutral component physical association physical association Fig 4B, Supp 6B