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 and characterization of an alternatively spliced isoform of the human protein phosphatase 2Aα catalytic subunit. Identification and characterization of an alternatively spliced isoform of the human protein phosphatase 2Aα catalytic subunit. The Journal of biological chemistry 2012 imex curation Migueleti DL., Smetana JH., Nunes HF., Kobarg J., Zanchin NI. Results from PP2A activity assays have not been added, as no purification after an initial IP was carried out, gels are not shown, so it is not possible to determine if contaminants were present. nilson.zanchin@pq.cnpq.br,jorg.kobarg@lnbio.org.br 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) pull down pull down anti tag western anti tag western blot anti tag western Migueleti DL., Smetana JH., Nunes HF., Kobarg J., Zanchin NI. this could be an "anti-Tag" Western, as they DO show clear bands for each component? Results from PP2A activity assays have not been added, as no purification after an initial IP was carried out, gels are not shown, so it is not possible to determine if contaminants were present. nilson.zanchin@pq.cnpq.br,jorg.kobarg@lnbio.org.br Accepted 2018-JUN-20 AT 13:26 BST AT 13:26 BST by PPORRAS The Journal of biological chemistry 2012 Only protein-protein interactions imex curation imex curation Identification and characterization of an alternatively spliced isoform of the human protein phosphatase 2Aα catalytic subunit. Identification and characterization of an alternatively spliced isoform of the human protein phosphatase 2Aα catalytic subunit. The Journal of biological chemistry 2012 imex curation Migueleti DL., Smetana JH., Nunes HF., Kobarg J., Zanchin NI. Results from PP2A activity assays have not been added, as no purification after an initial IP was carried out, gels are not shown, so it is not possible to determine if contaminants were present. nilson.zanchin@pq.cnpq.br,jorg.kobarg@lnbio.org.br 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 western blot western blot Immuno blot Immuno blot Migueleti DL., Smetana JH., Nunes HF., Kobarg J., Zanchin NI. N- terminal FLAG tag, as ALL pcDNA-FLAG constucts I found have N-term tag. Results from PP2A activity assays have not been added, as no purification after an initial IP was carried out, gels are not shown, so it is not possible to determine if contaminants were present. nilson.zanchin@pq.cnpq.br,jorg.kobarg@lnbio.org.br Accepted 2018-JUN-20 AT 13:26 BST AT 13:26 BST by PPORRAS The Journal of biological chemistry 2012 Only protein-protein interactions imex curation imex curation Identification and characterization of an alternatively spliced isoform of the human protein phosphatase 2Aα catalytic subunit. Identification and characterization of an alternatively spliced isoform of the human protein phosphatase 2Aα catalytic subunit. The Journal of biological chemistry 2012 imex curation Migueleti DL., Smetana JH., Nunes HF., Kobarg J., Zanchin NI. Results from PP2A activity assays have not been added, as no purification after an initial IP was carried out, gels are not shown, so it is not possible to determine if contaminants were present. nilson.zanchin@pq.cnpq.br,jorg.kobarg@lnbio.org.br 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Migueleti DL., Smetana JH., Nunes HF., Kobarg J., Zanchin NI. Results from PP2A activity assays have not been added, as no purification after an initial IP was carried out, gels are not shown, so it is not possible to determine if contaminants were present. this could be an "anti-Tag" Western, as they DO show clear bands for each component? Results from PP2A activity assays have not been added, as no purification after an initial IP was carried out, gels are not shown, so it is not possible to determine if contaminants were present. nilson.zanchin@pq.cnpq.br,jorg.kobarg@lnbio.org.br Accepted 2018-JUN-20 AT 13:26 BST AT 13:26 BST by PPORRAS The Journal of biological chemistry 2012 Only protein-protein interactions imex curation imex curation p67775-1 Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform Replication protein C PP2Acalpha1 PPP2CA protein protein human Homo sapiens Human MDEKVFTKELDQWIEQLNECKQLSESQVKSLCEKAKEILTKESNVQEVRCPVTVCGDVHGQFHDLMELFRIGGKSPDTNYLFMGDYVDRGYYSVETVTLLVALKVRYRERITILRGNHESRQITQVYGFYDECLRKYGNANVWKYFTDLFDYLPLTALVDGQIFCLHGGLSPSIDTLDHIRALDRLQEVPHEGPMCDLLWSDPDDRGGWGISPRGAGYTFGQDISETFNHANGLTLVSRAHQLVMEGYNWCHDRNVVTIFSAPNYCYRCGNQAAIMELDDTLKYSFLQFDPAPRRGEPHVTRRTPDYFL C602291F78F34555 p67775-2 Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform Replication protein C PP2Acalpha2 PPP2CA protein protein human Homo sapiens Human MDEKVFTKELDQWIEQLNECKQLSESQVKSLCEKAKEILTKESNVQEVRCPVTVCGDVHGQFHDLMELFRIGGKSPDTNYLFMGDYVDRGYYSVETVTLLVALKVRYRERITILRGNHESRQITQVYGFYDECLRKYGNANVWKYFTDLFDYLPLTALVDGQIFCLHGGLSPSIDTLDHIRALDRLQEVPHEGYNWCHDRNVVTIFSAPNYCYRCGNQAAIMELDDTLKYSFLQFDPAPRRGEPHVTRRTPDYFL Catalytically inactive, shows enhanced binding to IGBP1, and does not interact with the scaffolding subunit PPP2R1A. B50450F64CB286AB 2aaa_human Serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A alpha isoform PPP2R1A PP2A subunit A isoform PR65-alpha PP2A subunit A isoform R1-alpha Medium tumor antigen-associated 61 kDa protein protein protein human Homo sapiens Human MAAADGDDSLYPIAVLIDELRNEDVQLRLNSIKKLSTIALALGVERTRSELLPFLTDTIYDEDEVLLALAEQLGTFTTLVGGPEYVHCLLPPLESLATVEETVVRDKAVESLRAISHEHSPSDLEAHFVPLVKRLAGGDWFTSRTSACGLFSVCYPRVSSAVKAELRQYFRNLCSDDTPMVRRAAASKLGEFAKVLELDNVKSEIIPMFSNLASDEQDSVRLLAVEACVNIAQLLPQEDLEALVMPTLRQAAEDKSWRVRYMVADKFTELQKAVGPEITKTDLVPAFQNLMKDCEAEVRAAASHKVKEFCENLSADCRENVIMSQILPCIKELVSDANQHVKSALASVIMGLSPILGKDNTIEHLLPLFLAQLKDECPEVRLNIISNLDCVNEVIGIRQLSQSLLPAIVELAEDAKWRVRLAIIEYMPLLAGQLGVEFFDEKLNSLCMAWLVDHVYAIREAATSNLKKLVEKFGKEWAHATIIPKVLAMSGDPNYLHRMTTLFCINVLSEVCGQDITTKHMLPTVLRMAGDPVANVRFNVAKSLQKIGPILDNSTLQSEVKPILEKLTQDQDVDVKYFAQEALTVLSLA DIP protein P30153 original sequence version: 153 DIP protein P30153 original sequence version: 71 DIP protein P30153 original sequence version: 105 DIP protein P30153 original sequence version: 107 DIP protein P30153 original sequence version: 151 DIP protein P30153 original sequence version: 156 DIP protein P30153 original sequence version: 155 DIP protein P30153 original sequence version: 169 5174EBE94D537836 tiprl_human TIP41-like protein Putative MAPK-activating protein PM10 TIPRL Type 2A-interacting protein protein protein human Homo sapiens Human MMIHGFQSSHRDFCFGPWKLTASKTHIMKSADVEKLADELHMPSLPEMMFGDNVLRIQHGSGFGIEFNATDALRCVNNYQGMLKVACAEEWQESRTEGEHSKEVIKPYDWTYTTDYKGTLLGESLKLKVVPTTDHIDTEKLKAREQIKFFEEVLLFEDELHDHGVSSLSVKIRVMPSSFFLLLRFFLRIDGVLIRMNDTRLYHEADKTYMLREYTSRESKISSLMHVPPSLFTEPNEISQYLPIKEAVCEKLIFPERIDPNPADSQKSTQVE 23B730E0BC27DCEA ppme1_human Protein phosphatase methylesterase 1 PPME1 PME1 PP2593 PRO0750 protein protein human Homo sapiens Human MSALEKSMHLGRLPSRPPLPGSGGSQSGAKMRMGPGRKRDFSPVPWSQYFESMEDVEVENETGKDTFRVYKSGSEGPVLLLLHGGGHSALSWAVFTAAIISRVQCRIVALDLRSHGETKVKNPEDLSAETMAKDVGNVVEAMYGDLPPPIMLIGHSMGGAIAVHTASSNLVPSLLGLCMIDVVEGTAMDALNSMQNFLRGRPKTFKSLENAIEWSVKSGQIRNLESARVSMVGQVKQCEGITSPEGSKSIVEGIIEEEEEDEEGSESISKRKKEDDMETKKDHPYTWRIELAKTEKYWDGWFRGLSNLFLSCPIPKLLLLAGVDRLDKDLTIGQMQGKFQMQVLPQCGHAVHEDAPDKVAEAVATFLIRHRFAEPIGGFQCVFPGC 37B25324583E8578 igbp1_human Immunoglobulin-binding protein 1 IGBP1 IBP1 CD79a-binding protein 1 B-cell signal transduction molecule alpha 4 Renal carcinoma antigen NY-REN-16 Protein phosphatase 2/4/6 regulatory subunit protein protein human Homo sapiens Human MAAEDELQLPRLPELFETGRQLLDEVEVATEPAGSRIVQEKVFKGLDLLEKAAEMLSQLDLFSRNEDLEEIASTDLKYLLVPAFQGALTMKQVNPSKRLDHLQRAREHFINYLTQCHCYHVAEFELPKTMNNSAENHTANSSMAYPSLVAMASQRQAKIQRYKQKKELEHRLSAMKSAVESGQADDERVREYYLLHLQRWIDISLEEIESIDQEIKILRERDSSREASTSNSSRQERPPVKPFILTRNMAQAKVFGAGYPSLPTMTVSDWYEQHRKYGALPDQGIAKAAPEEFRKAAQQQEEQEEKEEEDDEQTLHRAREWDDWKDTHPRGYGNRQNMG BB00A116EB45273A lcmt1_human Leucine carboxyl methyltransferase 1 Protein-leucine O-methyltransferase LCMT1 LCMT [Phosphatase 2A protein]-leucine-carboxy methyltransferase 1 CGI-68 protein protein human Homo sapiens Human MATRQRESSITSCCSTSSCDADDEGVRGTCEDASLCKRFAVSIGYWHDPYIQHFVRLSKERKAPEINRGYFARVHGVSQLIKAFLRKTECHCQIVNLGAGMDTTFWRLKDEDLLPSKYFEVDFPMIVTRKLHSIKCKPPLSSPILELHSEDTLQMDGHILDSKRYAVIGADLRDLSELEEKLKKCNMNTQLPTLLIAECVLVYMTPEQSANLLKWAANSFERAMFINYEQVNMGDRFGQIMIENLRRRQCDLAGVETCKSLESQKERLLSNGWETASAVDMMELYNRLPRAEVSRIESLEFLDEMELLEQLMRHYCLCWATKGGNELGLKEITY C8D200118E6F353D ppp2ca-1 2 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range association association Figure 4C, 5G clone A4, C5. ppp2ca-igbp1-2 2 7 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range association association Figure 4C-2, 5G. clone 3.4 The interaction of PP2Aca2 with the PR65/A subunit was completely abolished, as was an interaction with PME1. the interaction of PP2Acα2 with the PR65/A subunit was completely abolished ppp2ca-tiprl-2 1 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus 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 c_terminus 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 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5A PME-1, TIPRL, and LCMT-1 all bind to both PP2Aca isoforms. ppp2ca-tiprl-3 1 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus 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 c_terminus 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 bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5Ai PME-1, TIPRL, and LCMT-1 all bind to both PP2Aca isoforms. ppp2ca-tiprl-4 1 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5B PME-1, TIPRL, and LCMT-1 all bind to both PP2Aca isoforms. ppp2ca-ppme1-1 1 8 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5B-2 PME-1, TIPRL, and LCMT-1 all bind to both PP2Aca isoforms. ppp2ca-ppme1-2 1 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5C PME-1, TIPRL, and LCMT-1 all bind to both PP2Aca isoforms. PME-1, TIPRL, and LCMT-1 all bind to both PP2Acα isoforms ppp2ca-lcmt1-1 1 10 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figure 5C-2 PME-1, TIPRL, and LCMT-1 all bind to both PP2Aca isoforms. PME-1, TIPRL, and LCMT-1 all bind to both PP2Acα isoforms ppp2ca-tiprl-5 3 9 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c_terminus 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 n_terminus 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus 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 c_terminus 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) 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Figure 5F TIPRL was detected in association with the PP2Ac·a4 complex; however, neither PME-1 nor LCMT-1 were detected. ppp2ca-tiprl-6 3 9 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate c_terminus 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 n_terminus 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus 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 c_terminus 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) 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed cell lysate cell lysate n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Figure 5E TIPRL was detected in association with the PP2Ac·a4 complex; however, neither PME-1 nor LCMT-1 were detected.