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/ MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. J. Biol. Chem. (0021-9258) 2003 imex curation Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer jfujita@virus.kyoto-u.ac.jp 07-Sept-2007: Contacted by JYOTI. Human placenta or a U-2 OS cDNA library orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis jfujita@virus.kyoto-u.ac.jp Accepted 2007-SEP-31 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. Human placenta or a U-2 OS cDNA library J. Biol. Chem. (0021-9258) 2003 orchard Only protein-protein interactions imex curation MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. J. Biol. Chem. (0021-9258) 2003 imex curation Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer jfujita@virus.kyoto-u.ac.jp 07-Sept-2007: Contacted by JYOTI. Human placenta or a U-2 OS cDNA library orchard Only protein-protein interactions human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis jfujita@virus.kyoto-u.ac.jp Accepted 2007-SEP-31 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 orchard Only protein-protein interactions imex curation MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. J. Biol. Chem. (0021-9258) 2003 imex curation Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer jfujita@virus.kyoto-u.ac.jp 07-Sept-2007: Contacted by JYOTI. Human placenta or a U-2 OS cDNA library orchard Only protein-protein interactions human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis jfujita@virus.kyoto-u.ac.jp Accepted 2007-SEP-31 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 orchard Only protein-protein interactions imex curation MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. J. Biol. Chem. (0021-9258) 2003 imex curation Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer jfujita@virus.kyoto-u.ac.jp 07-Sept-2007: Contacted by JYOTI. Human placenta or a U-2 OS cDNA library orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis jfujita@virus.kyoto-u.ac.jp Accepted 2007-SEP-31 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 orchard Only protein-protein interactions imex curation MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity. J. Biol. Chem. (0021-9258) 2003 imex curation Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer jfujita@virus.kyoto-u.ac.jp 07-Sept-2007: Contacted by JYOTI. Human placenta or a U-2 OS cDNA library orchard Only protein-protein interactions chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Nagao T., Higashitsuji H., Nonoguchi K., Sakurai T., Dawson S., Mayer RJ., Itoh K., Fujita J. Apoptosis - Interactions involving proteins with a function related to apoptosis jfujita@virus.kyoto-u.ac.jp Accepted 2007-SEP-31 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 orchard Only protein-protein interactions imex curation rb_human Retinoblastoma-associated protein pRb pp110 p105-Rb RB1 p110-RB1 protein protein human Homo sapiens Human MPPKTPRKTAATAAAAAAEPPAPPPPPPPEEDPEQDSGPEDLPLVRLEFEETEEPDFTALCQKLKIPDHVRERAWLTWEKVSSVDGVLGGYIQKKKELWGICIFIAAVDLDEMSFTFTELQKNIEISVHKFFNLLKEIDTSTKVDNAMSRLLKKYDVLFALFSKLERTCELIYLTQPSSSISTEINSALVLKVSWITFLLAKGEVLQMEDDLVISFQLMLCVLDYFIKLSPPMLLKEPYKTAVIPINGSPRTPRRGQNRSARIAKQLENDTRIIEVLCKEHECNIDEVKNVYFKNFIPFMNSLGLVTSNGLPEVENLSKRYEEIYLKNKDLDARLFLDHDKTLQTDSIDSFETQRTPRKSNLDEEVNVIPPHTPVRTVMNTIQQLMMILNSASDQPSENLISYFNNCTVNPKESILKRVKDIGYIFKEKFAKAVGQGCVEIGSQRYKLGVRLYYRVMESMLKSEEERLSIQNFSKLLNDNIFHMSLLACALEVVMATYSRSTSQNLDSGTDLSFPWILNVLNLKAFDFYKVIESFIKAEGNLTREMIKHLERCEHRIMESLAWLSDSPLFDLIKQSKDREGPTDHLESACPLNLPLQNNHTAADMYLSPVRSPKKKGSTTRVNSTANAETQATSAFQTQKPLKSTSLSLFYKKVYRLAYLRLNTLCERLLSEHPELEHIIWTLFQHTLQNEYELMRDRHLDQIMMCSMYGICKVKNIDLKFKIIVTAYKDLPHAVQETFKRVLIKEEEYDSIIVFYNSVFMQRLKTNILQYASTRPPTLSPIPHIPRSPYKFPSSPLRIPGGNIYISPLKSPYKISEGLPTPTKMTPRSRILVSIGESFGTSEKFQKINQMVCNSDRVLKRSAEGSNPPKPLKKLRFDIEGSDEADGSKHLPGESKFQQKLAEMTSTRTRMQKQKMNDSMDTSNKEEK DIP protein P06400 original sequence version: 102 DIP protein P06400 original sequence version: 195 DIP protein P06400 original sequence version: 204 DIP protein P06400 original sequence version: 126 DIP protein P06400 original sequence version: 151 DIP protein P06400 original sequence version: 153 DIP protein P06400 original sequence version: 181 DIP protein P06400 original sequence version: 184 DIP protein P06400 original sequence version: 213 DIP protein P06400 original sequence version: 208 DIP protein P06400 original sequence version: 174 DIP protein P06400 original sequence version: 212 DIP protein P06400 original sequence version: 179 DIP protein P06400 original sequence version: 146 C8E746111E19CC32 maga4_human Melanoma-associated antigen 4 MAGE-4 antigen MAGE-X2 antigen MAGE-41 antigen Cancer/testis antigen 1.4 MAGEA4 MAGE4 protein protein human Homo sapiens Human MSSEQKSQHCKPEEGVEAQEEALGLVGAQAPTTEEQEAAVSSSSPLVPGTLEEVPAAESAGPPQSPQGASALPTTISFTCWRQPNEGSSSQEEEGPSTSPDAESLFREALSNKVDELAHFLLRKYRAKELVTKAEMLERVIKNYKRCFPVIFGKASESLKMIFGIDVKEVDPASNTYTLVTCLGLSYDGLLGNNQIFPKTGLLIIVLGTIAMEGDSASEEEIWEELGVMGVYDGREHTVYGEPRKLLTQDWVQENYLEYRQVPGSNPARYEFLWGPRALAETSYVKVLEHVVRVNARVRIAYPSLREAALLEEEEGV C6AA2EE306963E7D prs6b_human 26S proteasome regulatory subunit 6B Proteasome 26S subunit ATPase 4 MB67-interacting protein PSMC4 TBP7 MIP224 Tat-binding protein 7 26S proteasome AAA-ATPase subunit RPT3 protein protein human Homo sapiens Human MEEIGILVEKAQDEIPALSVSRPQTGLSFLGPEPEDLEDLYSRYKKLQQELEFLEVQEEYIKDEQKNLKKEFLHAQEEVKRIQSIPLVIGQFLEAVDQNTAIVGSTTGSNYYVRILSTIDRELLKPNASVALHKHSNALVDVLPPEADSSIMMLTSDQKPDVMYADIGGMDIQKQEVREAVELPLTHFELYKQIGIDPPRGVLMYGPPGCGKTMLAKAVAHHTTAAFIRVVGSEFVQKYLGEGPRMVRDVFRLAKENAPAIIFIDEIDAIATKRFDAQTGADREVQRILLELLNQMDGFDQNVNVKVIMATNRADTLDPALLRPGRLDRKIEFPLPDRRQKRLIFSTITSKMNLSEEVDLEDYVARPDKISGADINSICQESGMLAVRENRYIVLAKDFEKAYKTVIKKDEQEHEFYK DIP protein P43686 original sequence version: 63 80F9523C61B88F0C psd10_human 26S proteasome non-ATPase regulatory subunit 10 26S proteasome regulatory subunit p28 Gankyrin PSMD10 p28(GANK) protein protein human Homo sapiens Human MEGCVSNLMVCNLAYSGKLEELKESILADKSLATRTDQDSRTALHWACSAGHTEIVEFLLQLGVPVNDKDDAGWSPLHIAASAGRDEIVKALLGKGAQVNAVNQNGCTPLHYAASKNRHEIAVMLLEGGANPDAKDHYEATAMHRAAAKGNLKMIHILLYYKASTNIQDTEGNTPLHLACDEERVEEAKLLVSQGASIYIENKEEKTPLQVAKGGLGLILKRMVEG DIP protein O75832 original sequence version: 149 57158E33146EC7C8 gankyrin-mage_a4 1 7 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role bait bait physical association physical association MAGE-A4 did not interact with the GAL4 DNA binding domain alone (Fig. 1A), indicating that it interacted with gankyrin. gankyrin-mage_a4-1 4 7 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 unspecified role unspecified role bait bait physical association physical association By contrast, a truncation of 10 amino acids from the C terminus of MAGE-A4 abolished its binding to gankyrin (Fig. 4B). The interaction was specific to MAGE-A4, because the corresponding regions of other MAGE proteins, although structurally quite similar to MAGE-A4, did not interact with gankyrin (Fig. 4, C and D). These results indicate that the C-terminal region of MAGE-A4 containing the HLA-A2-presented peptides (11, 12) (GVYDGREHTV and YLEYRQVPV) specifically interacts with gankyrin. 1A 4B gankyrin-mage_a4-2 5 7 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 9 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 physical association physical association When cell lysates were immunoprecipitated with an anti-FLAG antibody, HA-gankyrin was detected in them but not in precipitates from cells co-transfected with parental FLAG vector and HA-gankyrin 1B gankyrin-mage_a4-3 5 7 unspecified role unspecified role prey prey region 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 c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 cofactor cofactor coenzyme bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Reciprocally, truncated MAGE-A4 was detected in the anti-HA immunoprecipitates 1B gankyrin-mage_a4-4 2 7 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association the endogenous gankyrin was co-immunoprecipitated with MAGE proteins by anti-MAGE antibody. 2C gankyrin-rb 3 6 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 9 unspecified role unspecified role bait bait region 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 When U-2 OS cells were co-transfected with plasmids expressing FLAG-tagged gankyrin and HA-tagged Rb, HA-Rb was immunoprecipitated with anti-FLAG antibody as expected 3A gankyrin-psmc4 1 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association MAGE-A4 did not interact with the GAL4 DNA binding domain alone (Fig. 1A), indicating that it interacted with gankyrin. The authors state that the C-terminal region of the 26S protease regulatory subunit 6B protein binds to the bait protein , but do not give the coordinates of this region. gankyrin-psmc4-1 3 9 unspecified role unspecified role bait bait region 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 8 unspecified role unspecified role prey prey region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association When U-2 OS cells were co-transfected with plasmids expressing FLAG-tagged gankyrin and EGFP-S6 fusion protein, EGFP-S6 was immunoprecipitated with anti-FLAG antibody as expected 3B