bhf-ucl WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. Cell Res. (1001-0602) 2009 imex curation Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins yjin@sibs.ac.cn 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins yjin@sibs.ac.cn Accepted 2013-AUG-29 by ORCHARD 09-Sep-2013: Contacted by IntAct-Help. Cell Res. (1001-0602) 2009 imex curation WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. Cell Res. (1001-0602) 2009 imex curation Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins yjin@sibs.ac.cn 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro ubiquitinase assay ubiquitinase assay ubiquitinase assay polyclonal western polyclonal antibody western blot polyclonal western Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins yjin@sibs.ac.cn Accepted 2013-AUG-29 by ORCHARD 09-Sep-2013: Contacted by IntAct-Help. Cell Res. (1001-0602) 2009 imex curation WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. Cell Res. (1001-0602) 2009 imex curation Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins yjin@sibs.ac.cn 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins 1. figures 1A, 2B and 2C: participant detection method=coomassie blue so updated and kept under same experiment. 2. removed links. yjin@sibs.ac.cn Accepted 2013-AUG-29 by ORCHARD 09-Sep-2013: Contacted by IntAct-Help. Cell Res. (1001-0602) 2009 imex curation WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. WWP2 promotes degradation of transcription factor OCT4 in human embryonic stem cells. Cell Res. (1001-0602) 2009 imex curation Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins yjin@sibs.ac.cn 09-Sep-2013: Contacted by IntAct-Help. Only protein-protein interactions human-embryonic_stem_ cells Human-embryonic_stem_ cells (es cells) embryonic_stem_cell Embryonic stem cell (es cells) anti bait coip anti bait coimmunoprecipitation anti bait coip polyclonal western polyclonal antibody western blot polyclonal western Xu H., Wang W., Li C., Yu H., Yang A., Wang B., Jin Y. Cardiac - Interactions involving cardiac related proteins yjin@sibs.ac.cn Accepted 2013-AUG-29 by ORCHARD 09-Sep-2013: Contacted by IntAct-Help. Cell Res. (1001-0602) 2009 imex curation ub2e1_human Ubiquitin-conjugating enzyme E2 E1 UBE2E1 UBCH6 Ubiquitin-protein ligase E1 Ubiquitin carrier protein E1 UbcH6 (E3-independent) E2 ubiquitin-conjugating enzyme E1 E2 ubiquitin-conjugating enzyme E1 protein protein human Homo sapiens Human MSDDDSRASTSSSSSSSSNQQTEKETNTPKKKESKVSMSKNSKLLSTSAKRIQKELADITLDPPPNCSAGPKGDNIYEWRSTILGPPGSVYEGGVFFLDITFTPEYPFKPPKVTFRTRIYHCNINSQGVICLDILKDNWSPALTISKVLLSICSLLTDCNPADPLVGSIATQYMTNRAEHDRMARQWTKRYAT 2FBC50BE2A6A0008 po5f1_human POU domain, class 5, transcription factor 1 Octamer-binding transcription factor 3 Octamer-binding protein 3 POU5F1 OCT3 OCT4 OTF3 Octamer-binding protein 4 protein protein human Homo sapiens Human MAGHLASDFAFSPPPGGGGDGPGGPEPGWVDPRTWLSFQGPPGGPGIGPGVGPGSEVWGIPPCPPPYEFCGGMAYCGPQVGVGLVPQGGLETSQPEGEAGVGVESNSDGASPEPCTVTPGAVKLEKEKLEQNPEESQDIKALQKELEQFAKLLKQKRITLGYTQADVGLTLGVLFGKVFSQTTICRFEALQLSFKNMCKLRPLLQKWVEEADNNENLQEICKAETLVQARKRKRTSIENRVRGNLENLFLQCPKPTLQQISHIAQQLGLEKDVVRVWFCNRRQKGKRSSSDYAQREDFEAAGSPFSGGPVSFPLAPGPHFGTPGYGSPHFTALYSSVPFPEGEAFPPVSVTTLGSPMHSN DIP protein Q01860 original sequence version: 131 DIP protein Q01860 original sequence version: 150 DIP protein Q01860 original sequence version: 138 934C58DAEA0C535B wwp2_human NEDD4-like E3 ubiquitin-protein ligase WWP2 WW domain-containing protein 2 Atrophin-1-interacting protein 2 WWP2 HECT-type E3 ubiquitin transferase WWP2 protein protein human Homo sapiens Human MASASSSRAGVALPFEKSQLTLKVVSAKPKVHNRQPRINSYVEVAVDGLPSETKKTGKRIGSSELLWNEIIILNVTAQSHLDLKVWSCHTLRNELLGTASVNLSNVLKNNGGKMENMQLTLNLQTENKGSVVSGGELTIFLDGPTVDLGNVPNGSALTDGSQLPSRDSSGTAVAPENRHQPPSTNCFGGRSRTHRHSGASARTTPATGEQSPGARSRHRQPVKNSGHSGLANGTVNDEPTTATDPEEPSVVGVTSPPAAPLSVTPNPNTTSLPAPATPAEGEEPSTSGTQQLPAAAQAPDALPAGWEQRELPNGRVYYVDHNTKTTTWERPLPPGWEKRTDPRGRFYYVDHNTRTTTWQRPTAEYVRNYEQWQSQRNQLQGAMQHFSQRFLYQSSSASTDHDPLGPLPPGWEKRQDNGRVYYVNHNTRTTQWEDPRTQGMIQEPALPPGWEMKYTSEGVRYFVDHNTRTTTFKDPRPGFESGTKQGSPGAYDRSFRWKYHQFRFLCHSNALPSHVKISVSRQTLFEDSFQQIMNMKPYDLRRRLYIIMRGEEGLDYGGIAREWFFLLSHEVLNPMYCLFEYAGKNNYCLQINPASSINPDHLTYFRFIGRFIAMALYHGKFIDTGFTLPFYKRMLNKRPTLKDLESIDPEFYNSIVWIKENNLEECGLELYFIQDMEILGKVTTHELKEGGESIRVTEENKEEYIMLLTDWRFTRGVEEQTKAFLDGFNEVAPLEWLRYFDEKELELMLCGMQEIDMSDWQKSTIYRHYTKNSKQIQWFWQVVKEMDNEKRIRLLQFVTGTCRLPVGGFAELIGSNGPQKFCIDKVGKETWLPRSHTCFNRLDLPPYKSYEQLREKLLYAIEETEGFGQE FCCD75CBA61F2204 uba1_rabit Ubiquitin-like modifier-activating enzyme 1 Ubiquitin-activating enzyme E1 UBA1 UBE1 protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MSSSPLSKKRRVSGPDPKPGSNCSPAQSVLPQVPSAPTNGMAKNGSEADIDEGLYSRQLYVLGHEAMKRLQTSSVLVSGLRGLGVEIAKNIILGGVKAVTLHDQGTAQWADLSSQFYLREEDIGKNRAEVSQPRLAELNSYVPVTAYTGPLVEDFLSGFQVVVLTNSPLEDQLRVGEFCHSRGIKLVVADTRGLFGQLFCDFGEEMILTDSNGEQPLSTMVSMVTKDNPGVVTCLDEARHGFESGDFVSFSEVQGMTELNGNQPIEIKVLGPYTFSICDTSNFSDYIRGGIVSQVKVPKKISFKSLSASLAEPDFVMTDFAKFSRPAQLHIGFQALHKFCAQHSRPPRPRNEEDAAELVTLARAVNSKASSAVQQDSLDEDLIRNLAFVAAGDLAPINAFIGGLAAQEVMKACSGKFMPIMQWLYFDALECLPEDKESLTEDKCLPRQNRYDGQVAVFGSDLQEKLGRQKYFLVGAGAIGCELLKNFAMIGLGCGENGEIIVTDMDTIEKSNLNRQFLFRPWDVTKLKSDTAAAAVHQMNPHIRVTSHQNRVGPDTERIYDDDFFQTLDGVANALDNVDARMYMDRRCVYYRKPLLESGTLGTKGNVQVVIPFLTESYSSSQDPPEKSIPICTLKNFPNAIEHTLQWARDEFEGLFKQPAENVNQYLTDPKFVERTLRLAGTQPLEVLEAVQRSLVLQLPQSWADCVTWACHHWHTQYSNNIRQLLHNFPPDQLTSSGAPFWSGPKRCPHPLTFDVSNPLHLDYVMAAANLFAQTYGLAGSQDRAAVATLLQSVQVPEFTPKSGVKIHVSDQELQSANASVDDSRLEELKATLPSPDKLPGFKMYPIDFEKDDDSNFHMDFIVAASNLRAENYDIPPADRHKSKLIAGKIIPAIATTTAAVVGLVCLELYKVVQGHRHLDSYKNGFLNLALPFFGFSEPLAAPRHQYYNQEWTLWDRFEVQGLQPNGEEMTLKQFLDYFKTEHKLEITMLSQGVSMLYSFFMPAAKLKERLDQPMTEIVSRVSKRKLGRHVRALVLELCCNDESGEDVEVPYVRYTIR 1196EAC5EEEC06A5 wwp2-pou5f1 3 6 unspecified role unspecified role prey prey his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ww domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ww2 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ww3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ww4 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ww1 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) The authors state that ww2 domain is required (sufficient in the authors' terms) to bind to OCT4 based on its thick band in fig 2c and faint bands for the other three domains ww1, ww3 and ww4. But as the faint bands are clearly there, my interpretation was that each of the four domains is sufficient to bind. direct interaction direct interaction Figures: 1 A, 2 B and 2 C wwp2-pou5f1-1 1 7 unspecified role unspecified role bait bait ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey flag tag 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 Figure 1 B pou5f1-wwp2 4 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Figure 1 C pou5f1-uba1-2 2 8 ancillary ancillary neutral component neutral component 5 ancillary ancillary neutral component neutral component his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate neutral component neutral component his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 enzyme enzyme neutral component neutral component O00308:p.Cys838Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain C A Wed May 02 16:46:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys838ala' Tue Jun 30 20:08:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys838Ala' The single point mutation totally disrupts the interaction in context of an enzymatic reaction, hence used "mutation disrupting". gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) ubiquitination ubiquitination reaction ubiquitination Figure 3 A protein ubiquitination of pou5f1 E1 = Ubiquitin Activating Enzyme 1 Q29504 is cited in reference Xu et al., 2004. UbcH6 = Ubiquitin-conjugating enzyme E2 E1 = P51965.