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/ dadke-2007-2 Regulation of protein tyrosine phosphatase 1B by sumoylation. 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 tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Dadke S., Cotteret S., Yip SC., Jaffer ZM., Haj F., Ivanov A., Rauscher F., Shuai K., Ng T., Neel BG., Chernoff J. PTP1B-null MEFs or wild-type PTP1B MEFs were either transfected or not with HA-PIAS1. The author confirmed by email that the organism of origin of the recombinant proteins, PTP1B, PIAS1 and the insulin receptor, was human, and that the SUMO-1 construct was of murine origin. J_Chernoff@fccc.edu Nat. Cell Biol. (1465-7392) 2007 dadke-2007-3 Regulation of protein tyrosine phosphatase 1B by sumoylation. in vitro In vitro phosphatase assay phosphatase assay predetermined predetermined participant predetermined Dadke S., Cotteret S., Yip SC., Jaffer ZM., Haj F., Ivanov A., Rauscher F., Shuai K., Ng T., Neel BG., Chernoff J. PTP1B-null MEFs or wild-type PTP1B MEFs were either transfected or not with HA-PIAS1. The author confirmed by email that the organism of origin of the recombinant proteins, PTP1B, PIAS1 and the insulin receptor, was human, and that the SUMO-1 construct was of murine origin. J_Chernoff@fccc.edu Nat. Cell Biol. (1465-7392) 2007 dadke-2007-1 Regulation of protein tyrosine phosphatase 1B by sumoylation. 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 anti tag western anti tag western blot anti tag western Dadke S., Cotteret S., Yip SC., Jaffer ZM., Haj F., Ivanov A., Rauscher F., Shuai K., Ng T., Neel BG., Chernoff J. PTP1B-null MEFs or wild-type PTP1B MEFs were either transfected or not with HA-PIAS1. The author confirmed by email that the organism of origin of the recombinant proteins, PTP1B, PIAS1 and the insulin receptor, was human, and that the SUMO-1 construct was of murine origin. J_Chernoff@fccc.edu Nat. Cell Biol. (1465-7392) 2007 sumo1_mouse Small ubiquitin-related modifier 1 Sumo1 Smt3c Smt3h3 Ubl1 Ubiquitin-like protein SMT3C SMT3 homolog 3 Ubiquitin-homology domain protein PIC1 protein protein mouse Mus musculus Mouse MSDQEAKPSTEDLGDKKEGEYIKLKVIGQDSSEIHFKVKMTTHLKKLKESYCQRQGVPMNSLRFLFEGQRIADNHTPKELGMEEEDVIEVYQEQTGGHSTV ptn1_human Tyrosine-protein phosphatase non-receptor type 1 PTPN1 PTP1B Protein-tyrosine phosphatase 1B protein protein human Homo sapiens Human MEMEKEFEQIDKSGSWAAIYQDIRHEASDFPCRVAKLPKNKNRNRYRDVSPFDHSRIKLHQEDNDYINASLIKMEEAQRSYILTQGPLPNTCGHFWEMVWEQKSRGVVMLNRVMEKGSLKCAQYWPQKEEKEMIFEDTNLKLTLISEDIKSYYTVRQLELENLTTQETREILHFHYTTWPDFGVPESPASFLNFLFKVRESGSLSPEHGPVVVHCSAGIGRSGTFCLADTCLLLMDKRKDPSSVDIKKVLLEMRKFRMGLIQTADQLRFSYLAVIEGAKFIMGDSSVQDQWKELSHEDLEPPPEHIPPPPRPPKRILEPHNGKCREFFPNHQWVKEETQEDKDCPIKEEKGSPLNAAPYGIESMSQDTEVRSRVVGGSLRGAQAASPAKGEPSLPEKDEDHALSYWKPFLVNMCVATVLTAGAYLCYRFLFNSNT 4-nitrophenyl dihydrogen phosphate 4-nitrophenyl dihydrogen phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) insr_human Insulin receptor INSR protein protein human Homo sapiens Human MATGGRRGAAAAPLLVAVAALLLGAAGHLYPGEVCPGMDIRNNLTRLHELENCSVIEGHLQILLMFKTRPEDFRDLSFPKLIMITDYLLLFRVYGLESLKDLFPNLTVIRGSRLFFNYALVIFEMVHLKELGLYNLMNITRGSVRIEKNNELCYLATIDWSRILDSVEDNYIVLNKDDNEECGDICPGTAKGKTNCPATVINGQFVERCWTHSHCQKVCPTICKSHGCTAEGLCCHSECLGNCSQPDDPTKCVACRNFYLDGRCVETCPPPYYHFQDWRCVNFSFCQDLHHKCKNSRRQGCHQYVIHNNKCIPECPSGYTMNSSNLLCTPCLGPCPKVCHLLEGEKTIDSVTSAQELRGCTVINGSLIINIRGGNNLAAELEANLGLIEEISGYLKIRRSYALVSLSFFRKLRLIRGETLEIGNYSFYALDNQNLRQLWDWSKHNLTITQGKLFFHYNPKLCLSEIHKMEEVSGTKGRQERNDIALKTNGDQASCENELLKFSYIRTSFDKILLRWEPYWPPDFRDLLGFMLFYKEAPYQNVTEFDGQDACGSNSWTVVDIDPPLRSNDPKSQNHPGWLMRGLKPWTQYAIFVKTLVTFSDERRTYGAKSDIIYVQTDATNPSVPLDPISVSNSSSQIILKWKPPSDPNGNITHYLVFWERQAEDSELFELDYCLKGLKLPSRTWSPPFESEDSQKHNQSEYEDSAGECCSCPKTDSQILKELEESSFRKTFEDYLHNVVFVPRKTSSGTGAEDPRPSRKRRSLGDVGNVTVAVPTVAAFPNTSSTSVPTSPEEHRPFEKVVNKESLVISGLRHFTGYRIELQACNQDTPEERCSVAAYVSARTMPEAKADDIVGPVTHEIFENNVVHLMWQEPKEPNGLIVLYEVSYRRYGDEELHLCVSRKHFALERGCRLRGLSPGNYSVRIRATSLAGNGSWTEPTYFYVTDYLDVPSNIAKIIIGPLIFVFLFSVVIGSIYLFLRKRQPDGPLGPLYASSNPEYLSASDVFPCSVYVPDEWEVSREKITLLRELGQGSFGMVYEGNARDIIKGEAETRVAVKTVNESASLRERIEFLNEASVMKGFTCHHVVRLLGVVSKGQPTLVVMELMAHGDLKSYLRSLRPEAENNPGRPPPTLQEMIQMAAEIADGMAYLNAKKFVHRDLAARNCMVAHDFTVKIGDFGMTRDIYETDYYRKGGKGLLPVRWMAPESLKDGVFTTSSDMWSFGVVLWEITSLAEQPYQGLSNEQVLKFVMDGGYLDQPDNCPERVTDLMRMCWQFNPKMRPTFLEIVNLLKDDLHPSFPEVSFFHSEENKAPESEELEMEFEDMENVPLDRSSHCQREEAGGRDGGSSLGFKRSYEEHIPYTHMNGGKKNGRILTLPRSNPS pias1_human E3 SUMO-protein ligase PIAS1 Protein inhibitor of activated STAT protein 1 Gu-binding protein RNA helicase II-binding protein DEAD/H box-binding protein 1 PIAS1 DDXBP1 E3 SUMO-protein transferase PIAS1 protein protein human Homo sapiens Human MADSAELKQMVMSLRVSELQVLLGYAGRNKHGRKHELLTKALHLLKAGCSPAVQMKIKELYRRRFPQKIMTPADLSIPNVHSSPMPATLSPSTIPQLTYDGHPASSPLLPVSLLGPKHELELPHLTSALHPVHPDIKLQKLPFYDLLDELIKPTSLASDNSQRFRETCFAFALTPQQVQQISSSMDISGTKCDFTVQVQLRFCLSETSCPQEDHFPPNLCVKVNTKPCSLPGYLPPTKNGVEPKRPSRPINITSLVRLSTTVPNTIVVSWTAEIGRNYSMAVYLVKQLSSTVLLQRLRAKGIRNPDHSRALIKEKLTADPDSEIATTSLRVSLLCPLGKMRLTIPCRALTCSHLQCFDATLYIQMNEKKPTWVCPVCDKKAPYEHLIIDGLFMEILKYCTDCDEIQFKEDGTWAPMRSKKEVQEVSASYNGVDGCLSSTLEHQVASHHQSSNKNKKVEVIDLTIDSSSDEEEEEPSAKRTCPSLSPTSPLNNKGILSLPHQASPVSRTPSLPAVDTSYINTSLIQDYRHPFHMTPMPYDLQGLDFFPFLSGDNQHYNTSLLAAAAAAVSDDQDLLHSSRFFPYTSSQMFLDQLSAGGSTSLPTTNGSSSGSNSSLVSSNSLRESHSHTVTNRSSTDTASIFGIIPDIISLD ptpn1-sumo1 HA-PTP1B is sumoylated in MEFs transfected with HA-PTP1B and T7-SUMO-1 652898 N/A 652901 unspecified role unspecified role prey prey region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 652902 unspecified role unspecified role bait bait P18031:p.Lys335Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P18031:p.Lys347Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P18031:p.[Lys335Arg;Lys347Arg] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P18031:p.[Lys73Arg;Lys335Arg;Lys347Arg;Lys389Arg] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 P18031; Q93068 transfection 1 C, E, 3 A, B, C, S1, S2 PTP1B was not sumoylated in PIAS1-null MEFs but was sumoylated in wild-type PIAS1 MEFs, indicating that PIAS1 is required for sumoylation of PTP1B in fibroblasts. A nuclear-localized form (PTP-NLS) and a C-terminal truncation mutant (PTP-DeltaC; aa 1-403) were sumoylated to a much lesser degree, and plasma membrane-bound PTP1B was monosumoylated. This indicates that the endoplasmic reticulum localization of the wild type is required for maximal sumoylation of PTP1B. sumo1-ptp1b HA-PTP1B is sumoylated in MEFs transfected with HA-PTP1B and T7-SUMO-1 652898 N/A 652901 unspecified role unspecified role bait bait region t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 652902 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 false physical association physical association false false false P18031; P18031 transfection 1 B, S4 ptpn1-pnpp His-PTP1B dephosphorylates p-nitrophenol phosphate 652899 N/A 652903 enzyme target enzyme target substrate substrate unspecified role unspecified role N/A 652902 enzyme enzyme unspecified role unspecified role region 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 false ecoli Escherichia coli (strain K12) dephosphorylation dephosphorylation reaction dephosphorylation false false false 4 B, C, S3, S4 When p-nitrophenyl phosphate was used as substrate, wild-type and PTP1BKR rapidly hydrolysed this compound. Sumoylated wild-type PTP1B, in contrast, had only about 40% activity in this assay Sumoylation reduced the phosphatase activity of wild-type PTP1B to about 40% and treatment with SENP-1 (SUMO protease) increases activity. The catalytic activity of the sumoylation-resistant form of PTP1B was not adversely affected. ptpn1-insr His-PTP1B dephosphorylates the autophosphorylated beta-subunit of the insulin receptor 652899 N/A 652902 enzyme enzyme unspecified role unspecified role region 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 false P18031:p.[Lys73Arg;Lys335Arg;Lys347Arg;Lys389Arg] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 652904 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection dephosphorylation dephosphorylation reaction dephosphorylation false false false 4 D The insulin receptor precursor (P06213) contains the alpha and beta subunit. The vector used is stated to code for aa 978-1283. The UniProtKB entry states the beta subunit consists of aa 763-1382. Sumoylation of PTP1B strongly inhibited its phosphatase activity and this was reversed by SENP-1 treatment. The PTP(KR) sumoylation-resistant mutant was unaffected by the presence of sumoylation machinery or SUMO proteases. The insulin receptor beta subunit was dephosphorylated by PTP1B. ptpn1-pias1 PTP1B coimmunoprecipitates HA-PIAS1 in wild-type PTP1B MEF lysates 652900 N/A 652902 unspecified role unspecified role bait bait N/A 652905 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 false physical association physical association false false false 1 A, S4 O75925 transfection; P18031 infection