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 substrates of human protein-tyrosine phosphatase PTPN22. Identification of substrates of human protein-tyrosine phosphatase PTPN22. J. Biol. Chem. (0021-9258) 2006 imex curation Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com Accepted 2007-APR-12 by DAVET. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Identification of substrates of human protein-tyrosine phosphatase PTPN22. Identification of substrates of human protein-tyrosine phosphatase PTPN22. J. Biol. Chem. (0021-9258) 2006 imex curation Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down fingerprinting peptide massfingerprinting fingerprinting Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com Accepted 2007-APR-12 by DAVET. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Identification of substrates of human protein-tyrosine phosphatase PTPN22. Identification of substrates of human protein-tyrosine phosphatase PTPN22. J. Biol. Chem. (0021-9258) 2006 imex curation Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com 10-APR-2007: Contacted by JYOTI. 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 Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com Accepted 2007-APR-12 by DAVET. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Identification of substrates of human protein-tyrosine phosphatase PTPN22. Identification of substrates of human protein-tyrosine phosphatase PTPN22. J. Biol. Chem. (0021-9258) 2006 imex curation Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro phosphatase assay phosphatase assay predetermined predetermined participant predetermined Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com Accepted 2007-APR-12 by DAVET. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Identification of substrates of human protein-tyrosine phosphatase PTPN22. Identification of substrates of human protein-tyrosine phosphatase PTPN22. J. Biol. Chem. (0021-9258) 2006 imex curation Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com Accepted 2007-APR-12 by DAVET. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Identification of substrates of human protein-tyrosine phosphatase PTPN22. Identification of substrates of human protein-tyrosine phosphatase PTPN22. J. Biol. Chem. (0021-9258) 2006 imex curation Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com 10-APR-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Wu J., Katrekar A., Honigberg LA., Smith AM., Conn MT., Tang J., Jeffery D., Mortara K., Sampang J., Williams SR., Buggy J., Clark JM. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease 10exp9 Jurkat cells were treated with 100 uM pervanadate to stabilize tyrosine-phosphorylated proteins. Bait proteins coupled to streptavidin beads, prey proteins separated by PAGE prior to identification. lakedaly@yahoo.com Accepted 2007-APR-12 by DAVET. 10-APR-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation cd3z_human T-cell surface glycoprotein CD3 zeta chain T-cell receptor T3 zeta chain CD247 CD3Z T3Z TCRZ protein protein human Homo sapiens Human MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR 9408260374856EE9 zap70_human Tyrosine-protein kinase ZAP-70 ZAP70 SRK Syk-related tyrosine kinase 70 kDa zeta-chain associated protein protein protein human Homo sapiens Human MPDPAAHLPFFYGSISRAEAEEHLKLAGMADGLFLLRQCLRSLGGYVLSLVHDVRFHHFPIERQLNGTYAIAGGKAHCGPAELCEFYSRDPDGLPCNLRKPCNRPSGLEPQPGVFDCLRDAMVRDYVRQTWKLEGEALEQAIISQAPQVEKLIATTAHERMPWYHSSLTREEAERKLYSGAQTDGKFLLRPRKEQGTYALSLIYGKTVYHYLISQDKAGKYCIPEGTKFDTLWQLVEYLKLKADGLIYCLKEACPNSSASNASGAAAPTLPAHPSTLTHPQRRIDTLNSDGYTPEPARITSPDKPRPMPMDTSVYESPYSDPEELKDKKLFLKRDNLLIADIELGCGNFGSVRQGVYRMRKKQIDVAIKVLKQGTEKADTEEMMREAQIMHQLDNPYIVRLIGVCQAEALMLVMEMAGGGPLHKFLVGKREEIPVSNVAELLHQVSMGMKYLEEKNFVHRDLAARNVLLVNRHYAKISDFGLSKALGADDSYYTARSAGKWPLKWYAPECINFRKFSSRSDVWSYGVTMWEALSYGQKPYKKMKGPEVMAFIEQGKRMECPPECPPELYALMSDCWIYKWEDRPDFLTVEQRMRACYYSLASKVEGPPGSTQKAEAACA DIP protein P43403 original sequence version: 146 DIP protein P43403-1 original sequence version: 167 D1E1A8EC66FA116F vav_human Proto-oncogene vav VAV1 VAV protein protein human Homo sapiens Human MELWRQCTHWLIQCRVLPPSHRVTWDGAQVCELAQALRDGVLLCQLLNNLLPHAINLREVNLRPQMSQFLCLKNIRTFLSTCCEKFGLKRSELFEAFDLFDVQDFGKVIYTLSALSWTPIAQNRGIMPFPTEEESVGDEDIYSGLSDQIDDTVEEDEDLYDCVENEEAEGDEIYEDLMRSEPVSMPPKMTEYDKRCCCLREIQQTEEKYTDTLGSIQQHFLKPLQRFLKPQDIEIIFINIEDLLRVHTHFLKEMKEALGTPGAANLYQVFIKYKERFLVYGRYCSQVESASKHLDRVAAAREDVQMKLEECSQRANNGRFTLRDLLMVPMQRVLKYHLLLQELVKHTQEAMEKENLRLALDAMRDLAQCVNEVKRDNETLRQITNFQLSIENLDQSLAHYGRPKIDGELKITSVERRSKMDRYAFLLDKALLICKRRGDSYDLKDFVNLHSFQVRDDSSGDRDNKKWSHMFLLIEDQGAQGYELFFKTRELKKKWMEQFEMAISNIYPENATANGHDFQMFSFEETTSCKACQMLLRGTFYQGYRCHRCRASAHKECLGRVPPCGRHGQDFPGTMKKDKLHRRAQDKKRNELGLPKMEVFQEYYGLPPPPGAIGPFLRLNPGDIVELTKAEAEQNWWEGRNTSTNEIGWFPCNRVKPYVHGPPQDLSVHLWYAGPMERAGAESILANRSDGTFLVRQRVKDAAEFAISIKYNVEVKHIKIMTAEGLYRITEKKAFRGLTELVEFYQQNSLKDCFKSLDTTLQFPFKEPEKRTISRPAVGSTKYFGTAKARYDFCARDRSELSLKEGDIIKILNKKGQQGWWRGEIYGRVGWFPANYVEEDYSEYC AC3BC9736FD2F138 cd3e_human T-cell surface glycoprotein CD3 epsilon chain CD3E T3E T-cell surface antigen T3/Leu-4 epsilon chain protein protein human Homo sapiens Human MQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI A1603D01CE9957D7 tera_human Transitional endoplasmic reticulum ATPase VCP 15S Mg(2+)-ATPase p97 subunit Valosin-containing protein protein protein human Homo sapiens Human MASGADSKGDDLSTAILKQKNRPNRLIVDEAINEDNSVVSLSQPKMDELQLFRGDTVLLKGKKRREAVCIVLSDDTCSDEKIRMNRVVRNNLRVRLGDVISIQPCPDVKYGKRIHVLPIDDTVEGITGNLFEVYLKPYFLEAYRPIRKGDIFLVRGGMRAVEFKVVETDPSPYCIVAPDTVIHCEGEPIKREDEEESLNEVGYDDIGGCRKQLAQIKEMVELPLRHPALFKAIGVKPPRGILLYGPPGTGKTLIARAVANETGAFFFLINGPEIMSKLAGESESNLRKAFEEAEKNAPAIIFIDELDAIAPKREKTHGEVERRIVSQLLTLMDGLKQRAHVIVMAATNRPNSIDPALRRFGRFDREVDIGIPDATGRLEILQIHTKNMKLADDVDLEQVANETHGHVGADLAALCSEAALQAIRKKMDLIDLEDETIDAEVMNSLAVTMDDFRWALSQSNPSALRETVVEVPQVTWEDIGGLEDVKRELQELVQYPVEHPDKFLKFGMTPSKGVLFYGPPGCGKTLLAKAIANECQANFISIKGPELLTMWFGESEANVREIFDKARQAAPCVLFFDELDSIAKARGGNIGDGGGAADRVINQILTEMDGMSTKKNVFIIGATNRPDIIDPAILRPGRLDQLIYIPLPDEKSRVAILKANLRKSPVAKDVDLEFLAKMTNGFSGADLTEICQRACKLAIRESIESEIRRERERQTNPSAMEVEEDDPVPEIRRDHFEEAMRFARRSVSDNDIRKYEMFAQTLQQSRGFGSFRFPSGNQGGAGPSQGSGGGTGGSVYTEDNDDDLYG DIP protein P55072 original sequence version: 89 DIP protein P55072 original sequence version: 114 DIP protein P55072 original sequence version: 123 DIP protein P55072 original sequence version: 150 DIP protein P55072 original sequence version: 174 DIP protein P55072 original sequence version: 175 DIP protein P55072 original sequence version: 176 DIP protein P55072 original sequence version: 160 DIP protein P55072 original sequence version: 128 DIP protein P55072 original sequence version: 132 501B721D3A77BA8A lck_human Tyrosine-protein kinase Lck Protein YT16 Proto-oncogene Lck T cell-specific protein-tyrosine kinase LCK Lymphocyte cell-specific protein-tyrosine kinase p56-LCK Leukocyte C-terminal Src kinase protein protein human Homo sapiens Human MGCGCSSHPEDDWMENIDVCENCHYPIVPLDGKGTLLIRNGSEVRDPLVTYEGSNPPASPLQDNLVIALHSYEPSHDGDLGFEKGEQLRILEQSGEWWKAQSLTTGQEGFIPFNFVAKANSLEPEPWFFKNLSRKDAERQLLAPGNTHGSFLIRESESTAGSFSLSVRDFDQNQGEVVKHYKIRNLDNGGFYISPRITFPGLHELVRHYTNASDGLCTRLSRPCQTQKPQKPWWEDEWEVPRETLKLVERLGAGQFGEVWMGYYNGHTKVAVKSLKQGSMSPDAFLAEANLMKQLQHQRLVRLYAVVTQEPIYIITEYMENGSLVDFLKTPSGIKLTINKLLDMAAQIAEGMAFIEERNYIHRDLRAANILVSDTLSCKIADFGLARLIEDNEYTAREGAKFPIKWTAPEAINYGTFTIKSDVWSFGILLTEIVTHGRIPYPGMTNPEVIQNLERGYRMVRPDNCPEELYQLMRLCWKERPEDRPTFDYLRSVLEDFFTATEGQYQPQP DIP protein P06239 original sequence version: 164 DIP protein P06239-1 original sequence version: 193 44BFF0D43FFB420D ptn22_human Tyrosine-protein phosphatase non-receptor type 22 PTPN22 PTPN8 Hematopoietic cell protein-tyrosine phosphatase 70Z-PEP Lymphoid phosphatase PEST-domain phosphatase protein protein human Homo sapiens Human MDQREILQKFLDEAQSKKITKEEFANEFLKLKRQSTKYKADKTYPTTVAEKPKNIKKNRYKDILPYDYSRVELSLITSDEDSSYINANFIKGVYGPKAYIATQGPLSTTLLDFWRMIWEYSVLIIVMACMEYEMGKKKCERYWAEPGEMQLEFGPFSVSCEAEKRKSDYIIRTLKVKFNSETRTIYQFHYKNWPDHDVPSSIDPILELIWDVRCYQEDDSVPICIHCSAGCGRTGVICAIDYTWMLLKDGIIPENFSVFSLIREMRTQRPSLVQTQEQYELVYNAVLELFKRQMDVIRDKHSGTESQAKHCIPEKNHTLQADSYSPNLPKSTTKAAKMMNQQRTKMEIKESSSFDFRTSEISAKEELVLHPAKSSTSFDFLELNYSFDKNADTTMKWQTKAFPIVGEPLQKHQSLDLGSLLFEGCSNSKPVNAAGRYFNSKVPITRTKSTPFELIQQRETKEVDSKENFSYLESQPHDSCFVEMQAQKVMHVSSAELNYSLPYDSKHQIRNASNVKHHDSSALGVYSYIPLVENPYFSSWPPSGTSSKMSLDLPEKQDGTVFPSSLLPTSSTSLFSYYNSHDSLSLNSPTNISSLLNQESAVLATAPRIDDEIPPPLPVRTPESFIVVEEAGEFSPNVPKSLSSAVKVKIGTSLEWGGTSEPKKFDDSVILRPSKSVKLRSPKSELHQDRSSPPPPLPERTLESFFLADEDCMQAQSIETYSTSYPDTMENSTSSKQTLKTPGKSFTRSKSLKILRNMKKSICNSCPPNKPAESVQSNNSSSFLNFGFANRFSKPKGPRNPPPTWNI DIP protein Q9Y2R2 original sequence version: 69 1ABE8AE89C9D9FBF ptpn22-cd3e-1 2 11 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 8 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 9 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 12 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 10 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 13 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 Q9Y2R2:p.Asp195Ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 17:11:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp195ala' Tue Jun 30 16:29:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp195Ala' Q9Y2R2:p.Cys227Ser mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 14:32:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys227ser' Tue Jun 30 16:47:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys227Ser' n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) association association Identification of substrates of PTPN22. 1A-D ptpn22-cd3e-2 1 9 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 12 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 7 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 13 unspecified role unspecified role bait bait n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 Q9Y2R2:p.Asp195Ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 18:34:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp195ala' Tue Jun 30 19:58:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp195Ala' Q9Y2R2:p.Cys227Ser mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:09:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys227ser' Tue Jun 30 16:59:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys227Ser' ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia 8 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia association association proteins identified by mass spectrometry were confirmed in repeated substrate capture experiments by immunoblotting with antibodies specific for Zap70 (lanes 1 and 2), Lck (lanes 3 and 4), CD3{epsilon} (lanes 5 and 6), Vav (lanes 7 and 8), and phospho-TCR{zeta} (lanes 9 and 10). 1E ptpn22-lck-1 5 12 unspecified role unspecified role prey prey tyr394 (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 13 unspecified role unspecified role bait bait n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 Q9Y2R2:p.Asp195Ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 17:43:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp195ala' Tue Jun 30 15:47:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp195Ala' Q9Y2R2:p.Cys227Ser mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 17:33:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys227ser' Tue Jun 30 15:46:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys227Ser' ecoli Escherichia coli (strain K12) direct interaction direct interaction In contrast, activated Lck (phosphorylated at tyrosine 394) did interact with PTPN22cd, as ~30% of the activated Lck was pulled down by the substrate trap mutant (Fig. 2B, lanes 7-9). 2B, 2C ptpn22-zap70-1 5 8 unspecified role unspecified role prey prey 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 tyr-319 (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain tyr-493 (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain 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 13 unspecified role unspecified role bait bait n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 Q9Y2R2:p.Asp195Ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 16:12:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp195ala' Tue Jun 30 20:01:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp195Ala' Q9Y2R2:p.Cys227Ser mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:28:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys227ser' Tue Jun 30 19:21:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys227Ser' ecoli Escherichia coli (strain K12) direct interaction direct interaction In contrast, ~70% of the phosphorylated Zap70 was captured by the DACS mutant (Fig. 2E, lanes 4-6), demonstrating that PTPN22cd binds directly to Zap70 in an interaction that requires phosphorylation at one or both tyrosines. 2E lck-1 6 12 putative self putative self putative self putative self n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 phosphorylation phosphorylation reaction phosphorylation true Phosphorylation of LCK (P06239) at Tyr-394 Following autoactivation in the presence of ATP and Mg2+, tyrosine 394 became phosphorylated, whereas tyrosine 505 was not significantly affected (Fig. 2A). 2A P06239 is required for phosphorylation of P06239 lck-zap70-1 6 12 enzyme enzyme unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 8 enzyme target enzyme target substrate substrate unspecified role unspecified role 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 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 phosphorylation phosphorylation reaction phosphorylation Phosphorylation of ZAP70 (P43403) at Tyr-319 and Tyr-493 After activation of Zap70 by recombinant Lck, both sites became phosphorylated (Fig. 2D). 2D P06239 is required for phosphorylation of P43403 ptpn22-lck-2 4 12 enzyme target enzyme target substrate substrate unspecified role unspecified role tyr394 (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 13 enzyme enzyme unspecified role unspecified role n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobd Escherichia coli (strain B / BL21-DE3) dephosphorylation dephosphorylation reaction dephosphorylation Wild type PTPN22cd clearly dephosphorylated phosphotyrosine 394 of Lck, but did not dephosphorylate phosphotyrosine 505 of Lck (Fig. 3A). 3A Dephosphorylation of LCK (P06239) on Tyr-394 Q9Y2R2 is required for dephosphorylation of P06239 ptpn22-zap70-2 4 8 enzyme target enzyme target substrate substrate unspecified role unspecified role tyr-493 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain 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 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 13 enzyme enzyme unspecified role unspecified role n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecobd Escherichia coli (strain B / BL21-DE3) dephosphorylation dephosphorylation reaction dephosphorylation Interestingly, PTPN22cd dephosphorylated phosphotyrosine 493 (Fig. 3C, lanes 1-5) but not phosphotyrosine 319 (Fig. 3C, lanes 6-10). 3C Dephosphorylation of ZAP70 (P43403) on Tyr-394 Q9Y2R2 is required for dephosphorylation of P43403 ptpn22-cd247-1 5 13 unspecified role unspecified role bait bait n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 Q9Y2R2:p.Asp195Ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:42:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp195ala' Tue Jun 30 15:49:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp195Ala' Q9Y2R2:p.Cys227Ser mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 20:19:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys227ser' Tue Jun 30 17:03:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys227Ser' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey human-jurkat Homo sapiens jurkat Human T cell leukemia physical association physical association First, we tested the ability of various substrate trap mutants of PTPN22cd including D195A, C227S, D195A/Q274A, and DACS to pull down TCR{zeta} from Jurkat cell lysates. 4A In the presence of the phosphatase inhibitor, sodium vanadate, neither the wild type PTPN22cd nor the D195A mutant interacted with TCRzeta whereas in the absence of vanadate, only the D195A mutant could pull down TCRzeta lck-cd247-1 6 7 enzyme target enzyme target substrate substrate unspecified role unspecified role cytoplasmic region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 12 enzyme enzyme unspecified role unspecified role n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 phosphorylation phosphorylation reaction phosphorylation Phosphorylation of CD247 (P20963) To confirm that the interaction between PTPN22cd and TCR{zeta} was direct, recombinant cytoplasmic domain TCR{zeta} (cyto-TCR{zeta}) containing all three ITAMs was phosphorylated in vitro with purified Lck and tested as a potential substrate for PTPN22cd. 4C P06239 is required for phosphorylation of P20963 ptpn22-cd247-2 5 13 unspecified role unspecified role bait bait n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 Q9Y2R2:p.Asp195Ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:49:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp195ala' Tue Jun 30 20:24:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp195Ala' Q9Y2R2:p.Cys227Ser mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:03:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys227ser' Tue Jun 30 16:03:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys227Ser' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey cytoplasmic region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 106 101 direct interaction direct interaction In contrast, ~54% of the phosphorylated cyto-TCR{zeta} was captured by the DACS mutant, whereas none was pulled down by wild type PTPN22cd (Fig. 4C, lanes 4-6), 4D ptpn22-cd247-3 3 7 unspecified role unspecified role prey prey region (required) optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-jurkat Homo sapiens jurkat Human T cell leukemia 13 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range Q9Y2R2:p.Asp195Ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A Wed May 02 14:53:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp195ala' Tue Jun 30 19:25:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp195Ala' Q9Y2R2:p.Cys227Ser mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S Wed May 02 18:55:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys227ser' Tue Jun 30 16:16:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys227Ser' ecoli Escherichia coli (strain K12) physical association physical association However, only the DACS mutant of PTPN22 co-immunoprecipitated significant amounts of phosphorylated TCR{zeta} (seen as multiple bands on the immunoblot; Fig. 5A, lanes 1 and 2), indicating that the substrate-trapping mutant of full-length PTPN22 interacts with TCR{zeta} in transfected cells. 5A