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/ Tiki1 Is Required for Head Formation via Wnt Cleavage-Oxidation and Inactivation. Tiki1 Is Required for Head Formation via Wnt Cleavage-Oxidation and Inactivation. Cell (0092-8674) 2012 imex curation Zhang X., Abreu J.G., Yokota C., Macdonald B.T., Singh S., Coburn K.L., Cheong S.M., Zhang M.M., Ye Q.Z., Hang H.C., Steen H., He X. I have not captured the oligomerisation data, as it appears to be performed with unpurified cell lysate so a mixture of proteins could be binding and causing the shift. Additional details on constructs supplied by the authors. xi.he@childrens.harvard.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions 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 Zhang X., Abreu J.G., Yokota C., Macdonald B.T., Singh S., Coburn K.L., Cheong S.M., Zhang M.M., Ye Q.Z., Hang H.C., Steen H., He X. I have not captured the oligomerisation data, as it appears to be performed with unpurified cell lysate so a mixture of proteins could be binding and causing the shift. Additional details on constructs supplied by the authors. xi.he@childrens.harvard.edu Accepted 2012-AUG-02 by PPORRAS 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Tiki1 Is Required for Head Formation via Wnt Cleavage-Oxidation and Inactivation. Tiki1 Is Required for Head Formation via Wnt Cleavage-Oxidation and Inactivation. Cell (0092-8674) 2012 imex curation Zhang X., Abreu J.G., Yokota C., Macdonald B.T., Singh S., Coburn K.L., Cheong S.M., Zhang M.M., Ye Q.Z., Hang H.C., Steen H., He X. I have not captured the oligomerisation data, as it appears to be performed with unpurified cell lysate so a mixture of proteins could be binding and causing the shift. Additional details on constructs supplied by the authors. xi.he@childrens.harvard.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protease assay protease assay western blot western blot Immuno blot Immuno blot Zhang X., Abreu J.G., Yokota C., Macdonald B.T., Singh S., Coburn K.L., Cheong S.M., Zhang M.M., Ye Q.Z., Hang H.C., Steen H., He X. I have not captured the oligomerisation data, as it appears to be performed with unpurified cell lysate so a mixture of proteins could be binding and causing the shift. Additional details on constructs supplied by the authors. xi.he@childrens.harvard.edu Accepted 2012-AUG-02 by PPORRAS 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Tiki1 Is Required for Head Formation via Wnt Cleavage-Oxidation and Inactivation. Tiki1 Is Required for Head Formation via Wnt Cleavage-Oxidation and Inactivation. Cell (0092-8674) 2012 imex curation Zhang X., Abreu J.G., Yokota C., Macdonald B.T., Singh S., Coburn K.L., Cheong S.M., Zhang M.M., Ye Q.Z., Hang H.C., Steen H., He X. I have not captured the oligomerisation data, as it appears to be performed with unpurified cell lysate so a mixture of proteins could be binding and causing the shift. Additional details on constructs supplied by the authors. xi.he@childrens.harvard.edu 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions 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 Zhang X., Abreu J.G., Yokota C., Macdonald B.T., Singh S., Coburn K.L., Cheong S.M., Zhang M.M., Ye Q.Z., Hang H.C., Steen H., He X. I have not captured the oligomerisation data, as it appears to be performed with unpurified cell lysate so a mixture of proteins could be binding and causing the shift. Additional details on constructs supplied by the authors. xi.he@childrens.harvard.edu Accepted 2012-AUG-02 by PPORRAS 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation wnt3a_human Protein Wnt-3a WNT3A protein protein human Homo sapiens Human MAPLGYFLLLCSLKQALGSYPIWWSLAVGPQYSSLGSQPILCASIPGLVPKQLRFCRNYVEIMPSVAEGIKIGIQECQHQFRGRRWNCTTVHDSLAIFGPVLDKATRESAFVHAIASAGVAFAVTRSCAEGTAAICGCSSRHQGSPGKGWKWGGCSEDIEFGGMVSREFADARENRPDARSAMNRHNNEAGRQAIASHMHLKCKCHGLSGSCEVKTCWWSQPDFRAIGDFLKDKYDSASEMVVEKHRESRGWVETLRPRYTYFKVPTERDLVYYEASPNFCEPNPETGSFGTRDRTCNVSSHGIDGCDLLCCGRGHNARAERRREKCRCVFHWCCYVSCQECTRVYDVHTCK DIP protein P56704 original sequence version: 81 DIP protein P56704 original sequence version: 97 DIP protein P56704 original sequence version: 135 A317BD6D4A73920B wnt5a_mouse Protein Wnt-5a Wnt5a Wnt-5a protein protein mouse Mus musculus Mouse MKKPIGILSPGVALGTAGGAMSSKFFLMALATFFSFAQVVIEANSWWSLGMNNPVQMSEVYIIGAQPLCSQLAGLSQGQKKLCHLYQDHMQYIGEGAKTGIKECQYQFRHRRWNCSTVDNTSVFGRVMQIGSRETAFTYAVSAAGVVNAMSRACREGELSTCGCSRAARPKDLPRDWLWGGCGDNIDYGYRFAKEFVDARERERIHAKGSYESARILMNLHNNEAGRRTVYNLADVACKCHGVSGSCSLKTCWLQLADFRKVGDALKEKYDSAAAMRLNSRGKLVQVNSRFNSPTTQDLVYIDPSPDYCVRNESTGSLGTQGRLCNKTSEGMDGCELMCCGRGYDQFKTVQTERCHCKFHWCCYVKCKKCTEIVDQFVCK DIP protein P22725 original sequence version: 90 64CA36C516D22D8A tiki2_human Metalloprotease TIKI2 HKAT Heart, kidney and adipose-enriched transmembrane protein homolog TRABD2B TIKI2 TRAB domain-containing protein 2B protein protein human Homo sapiens Human MHAALAGPLLAALLATARARPQPPDGGQCRPPGSQRDLNSFLWTIRRDPPAYLFGTIHVPYTRVWDFIPDNSKAAFQASTRVYFELDLTDPYTISALASCQLLPHGENLQDVLPHELYWRLKRHLDYVKLMMPSWMTPAQRGKGLYADYLFNAIAGNWERKRPVWVMLMVNSLTERDVRFRGVPVLDLYLAQQAEKMKKTTGAVEQVEEQCHPLNNGLNFSQVLFALNQTLLQQESVRAGSLQASYTTEDLIKHYNCGDLSAVIFNHDTSQLPNFINTTLPPHEQVTAQEIDSYFRQELIYKRNERMGKRVMALLRENEDKICFFAFGAGHFLGNNTVIDILRQAGLEVDHTPAGQAIHSPAPQSPAPSPEGTSTSPAPVTPAAAVPEAPSVTPTAPPEDEDPALSPHLLLPDSLSQLEEFGRQRKWHKRQSTHQRPRQFNDLWVRIEDSTTASPPPLPLQPTHSSGTAKPPFQLSDQLQQQDPPGPASSSAPTLGLLPAIATTIAVCFLLHSLGPS E06AC08A4CC61840 lrp6_human Low-density lipoprotein receptor-related protein 6 LRP6 protein protein human Homo sapiens Human MGAVLRSLLACSFCVLLRAAPLLLYANRRDLRLVDATNGKENATIVVGGLEDAAAVDFVFSHGLIYWSDVSEEAIKRTEFNKTESVQNVVVSGLLSPDGLACDWLGEKLYWTDSETNRIEVSNLDGSLRKVLFWQELDQPRAIALDPSSGFMYWTDWGEVPKIERAGMDGSSRFIIINSEIYWPNGLTLDYEEQKLYWADAKLNFIHKSNLDGTNRQAVVKGSLPHPFALTLFEDILYWTDWSTHSILACNKYTGEGLREIHSDIFSPMDIHAFSQQRQPNATNPCGIDNGGCSHLCLMSPVKPFYQCACPTGVKLLENGKTCKDGATELLLLARRTDLRRISLDTPDFTDIVLQLEDIRHAIAIDYDPVEGYIYWTDDEVRAIRRSFIDGSGSQFVVTAQIAHPDGIAVDWVARNLYWTDTGTDRIEVTRLNGTMRKILISEDLEEPRAIVLDPMVGYMYWTDWGEIPKIERAALDGSDRVVLVNTSLGWPNGLALDYDEGKIYWGDAKTDKIEVMNTDGTGRRVLVEDKIPHIFGFTLLGDYVYWTDWQRRSIERVHKRSAEREVIIDQLPDLMGLKATNVHRVIGSNPCAEENGGCSHLCLYRPQGLRCACPIGFELISDMKTCIVPEAFLLFSRRADIRRISLETNNNNVAIPLTGVKEASALDFDVTDNRIYWTDISLKTISRAFMNGSALEHVVEFGLDYPEGMAVDWLGKNLYWADTGTNRIEVSKLDGQHRQVLVWKDLDSPRALALDPAEGFMYWTEWGGKPKIDRAAMDGSERTTLVPNVGRANGLTIDYAKRRLYWTDLDTNLIESSNMLGLNREVIADDLPHPFGLTQYQDYIYWTDWSRRSIERANKTSGQNRTIIQGHLDYVMDILVFHSSRQSGWNECASSNGHCSHLCLAVPVGGFVCGCPAHYSLNADNRTCSAPTTFLLFSQKSAINRMVIDEQQSPDIILPIHSLRNVRAIDYDPLDKQLYWIDSRQNMIRKAQEDGSQGFTVVVSSVPSQNLEIQPYDLSIDIYSRYIYWTCEATNVINVTRLDGRSVGVVLKGEQDRPRAVVVNPEKGYMYFTNLQERSPKIERAALDGTEREVLFFSGLSKPIALALDSRLGKLFWADSDLRRIESSDLSGANRIVLEDSNILQPVGLTVFENWLYWIDKQQQMIEKIDMTGREGRTKVQARIAQLSDIHAVKELNLQEYRQHPCAQDNGGCSHICLVKGDGTTRCSCPMHLVLLQDELSCGEPPTCSPQQFTCFTGEIDCIPVAWRCDGFTECEDHSDELNCPVCSESQFQCASGQCIDGALRCNGDANCQDKSDEKNCEVLCLIDQFRCANGQCIGKHKKCDHNVDCSDKSDELDCYPTEEPAPQATNTVGSVIGVIVTIFVSGTVYFICQRMLCPRMKGDGETMTNDYVVHGPASVPLGYVPHPSSLSGSLPGMSRGKSMISSLSIMGGSSGPPYDRAHVTGASSSSSSSTKGTYFPAILNPPPSPATERSHYTMEFGYSSNSPSTHRSYSYRPYSYRHFAPPTTPCSTDVCDSDYAPSRRMTSVATAKGYTSDLNYDSEPVPPPPTPRSQYLSAEENYESCPPSPYTERSYSHHLYPPPPSPCTDSS DIP protein O75581 original sequence version: 116 DIP protein O75581 original sequence version: 95 DIP protein O75581 original sequence version: 131 DIP protein O75581 original sequence version: 112 DIP protein O75581 original sequence version: 136 DIP protein O75581 original sequence version: 59 DIP protein O75581 original sequence version: 103 DIP protein O75581 original sequence version: 100 413D2CF70A5D8B5C wn11b_xenla Protein Wnt-11b wnt11b wnt-11 wnt11 Protein Wnt-11 protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MAPTRHWVTPLLLLCCSGICGAIQWLGLTVNGSRVAWNESEHCRLLDGLVPDQSQLCKRNLELMQSVVNAAKQTKLTCQMTLSDMRWNCSSVENAPSFTPDLSKGTRESAFVYALASATLSHTIARACASGELPTCSCGATPAEVPGTGFRWGGCGDNLHYGLNMGSAFVDAPMKSSKSAGTQATKIMNLHNNAVGRQVLMDSLETKCKCHGVSGSCSVKTCWKGLQDLPHIANELKSKYLGATKVIHRQTGTRRQLVPRELDIRPVRESELVYLVSSPDYCTKNPKLGSYGTQDRLCNKTSVGSDSCNLMCCGRGYNAYTETIVERCQCKYHWCCYVMCKKCERTVERYVCK B59609AB88915554 wnt8_xenla Protein Wnt-8 wnt8 protein protein xenla Xenopus laevis (African clawed frog) African clawed frog MQNTTLFILATLLIFCPFFTASAWSVNNFLMTGPKAYLTYSASVAVGAQNGIEECKYQFAWERWNCPESTLQLATHNGLRSATRETSFVHAISSAGVMYTLTRNCSMGDFDNCGCDDSRNGRIGGRGWVWGGCSDNAEFGERISKLFVDGLETGQDARALMNLHNNEAGRLAVKETMKRTCKCHGISGSCSIQTCWLQLAEFRDIGNHLKIKHDQALKLEMDKRKMRSGNSADNRGAIADAFSSVAGSELIFLEDSPDYCLKNISLGLQGTEGRECLQSGKNLSQWERRSCKRLCTDCGLRVEEKKTEIISSCNCKFHWCCTVKCEQCKQVVIKHFCARRERDSNMLNTKRKNRGHRR 8BC8B9E20016E504 wnt3a_mouse Protein Wnt-3a Wnt3a Wnt-3a protein protein mouse Mus musculus Mouse MAPLGYLLVLCSLKQALGSYPIWWSLAVGPQYSSLSTQPILCASIPGLVPKQLRFCRNYVEIMPSVAEGVKAGIQECQHQFRGRRWNCTTVSNSLAIFGPVLDKATRESAFVHAIASAGVAFAVTRSCAEGSAAICGCSSRLQGSPGEGWKWGGCSEDIEFGGMVSREFADARENRPDARSAMNRHNNEAGRQAIASHMHLKCKCHGLSGSCEVKTCWWSQPDFRTIGDFLKDKYDSASEMVVEKHRESRGWVETLRPRYTYFKVPTERDLVYYEASPNFCEPNPETGSFGTRDRTCNVSSHGIDGCDLLCCGRGHNARTERRREKCHCVFHWCCYVSCQECTRVYDVHTCK DIP protein P27467 original sequence version: 126 DIP protein P27467 original sequence version: 101 DIP protein P27467 original sequence version: 121 DIP protein P27467 original sequence version: 140 DIP protein P27467 original sequence version: 109 7ADFC5B38A8EFF63 fzd8_mouse Frizzled-8 Fzd8 protein protein mouse Mus musculus Mouse MEWGYLLEVTSLLAALAVLQRSSGAAAASAKELACQEITVPLCKGIGYNYTYMPNQFNHDTQDEAGLEVHQFWPLVEIQCSPDLKFFLCSMYTPICLEDYKKPLPPCRSVCERAKAGCAPLMRQYGFAWPDRMRCDRLPEQGNPDTLCMDYNRTDLTTAAPSPPRRLPPPPPPGEQPPSGSGHSRPPGARPPHRGGSSRGSGDAAAAPPSRGGKARPPGGGAAPCEPGCQCRAPMVSVSSERHPLYNRVKTGQIANCALPCHNPFFSQDERAFTVFWIGLWSVLCFVSTFATVSTFLIDMERFKYPERPIIFLSACYLFVSVGYLVRLVAGHEKVACSGGAPGAGGAGGAGGAAAAGAGAAGAGASSPGARGEYEELGAVEQHVRYETTGPALCTVVFLLVYFFGMASSIWWVILSLTWFLAAGMKWGNEAIAGYSQYFHLAAWLVPSVKSIAVLALSSVDGDPVAGICYVGNQSLDNLRGFVLAPLVIYLFIGTMFLLAGFVSLFRIRSVIKQQGGPTKTHKLEKLMIRLGLFTVLYTVPAAVVVACLFYEQHNRPRWEATHNCPCLRDLQPDQARRPDYAVFMLKYFMCLVVGITSGVWVWSGKTLESWRALCTRCCWASKGAAVGAGAGGSGPGGSGPGPGGGGGHGGGGGSLYSDVSTGLTWRSGTASSVSYPKQMPLSQV DIP protein Q61091 original sequence version: 123 DIP protein Q61091 original sequence version: 119 DIP protein Q61091 original sequence version: 108 9DB95F068BC45CD8 lrp6-wnt3a 3 7 unspecified role unspecified role bait bait extracellular region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey physical association physical association Fig. 4L Interaction not observed in TIKI2-expressing cells fzd8-wnt3a 3 11 unspecified role unspecified role bait bait region fc-igg tag fc-igg tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey physical association physical association Fig. 4L Interaction not observed in TIKI2-expressing cells wnt3a-u632a-1 2 10 enzyme target enzyme target substrate substrate unspecified role unspecified role region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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. 6 enzyme enzyme unspecified role unspecified role c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term 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 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain extracellular region sufficient to bind sufficient binding region sufficient to bind range range range range 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. protein cleavage protein cleavage Fig. 5O, 5P, 5Q, 5R Supp. FIgs. 7A- 7E 1,10-phenanthroline, (a divalent metal chelator), EDTA (a divalent metal chelator), Ni2+, Cu2+, Zn2+, Co2+, Mn2+ Cleavage of HA-Wnt3a became obvious at 2 hr and exhibited preference for neutral to slightly basic pH (pH 7–9). TIKI2N was insensitive to Bestatin, an aminopeptidase inhibitor, and to a mixture of inhibitors for serine, cysteine, and aspartic proteases and aminopeptidases. This suggests Tiki is probably a Co2+- or Mn2+-dependent metalloprotease. u632a-wnt3a-1 3 10 unspecified role unspecified role prey prey 6 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 c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range extracellular region sufficient to bind sufficient binding region sufficient to bind range range range range physical association physical association Supp. FIg. 6D u632a-wnt8 1 9 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 6 unspecified role unspecified role bait bait c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range extracellular region 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 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. physical association physical association Supp. FIg. 6E u632a-wnt5a 3 6 unspecified role unspecified role bait bait extracellular region sufficient to bind sufficient binding region sufficient to bind range range range range c-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term 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 5 unspecified role unspecified role prey prey physical association physical association Supp. FIg. 6F u632a-wnt11b 1 8 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 6 unspecified role unspecified role bait bait extracellular region 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 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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. physical association physical association Supp. FIg. 6G