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/ taira-2014-1 Phosphorylation of Grb14 BPS domain by GSK-3 correlates with complex forming of Grb14 and insulin receptor. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Taira J., Higashimoto Y. Diabetes - Interactions investigated in the context of Diabetes taira_junichi@med.kurume-u.ac.jp J. Biochem. (0021-924X) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. taira-2014-2 Phosphorylation of Grb14 BPS domain by GSK-3 correlates with complex forming of Grb14 and insulin receptor. 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 Taira J., Higashimoto Y. Diabetes - Interactions investigated in the context of Diabetes IR cDNA is supplied by Kazusa DNA Research Institute (pFN21AE3464). The ORF corresponds to a protein of 1370aa, corresponding to Uniprot ID P06213-2 . GRB14 cDNA, protein_id="BAG35971.1" has 99% homology to the reviewed UniProt entry ID= Q14449. taira_junichi@med.kurume-u.ac.jp J. Biochem. (0021-924X) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. taira-2014-3 Phosphorylation of Grb14 BPS domain by GSK-3 correlates with complex forming of Grb14 and insulin receptor. in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Taira J., Higashimoto Y. Diabetes - Interactions investigated in the context of Diabetes Kinase-Glo Luminescent Kinase Assay (Promega);quantitating the amount of ATP remaining in solution following a kinase reaction. The luminescent signal is correlated with the amount of ATP present and is inversely correlated with the amount of kinase activity. IR cDNA is supplied by Kazusa DNA Research Institute (pFN21AE3464). The ORF corresponds to a protein of 1370aa, corresponding to Uniprot ID P06213-2 . GRB14 cDNA, protein_id="BAG35971.1" has 99% homology to the sequence given in the reviewed UniProt entry ID= Q14449. Recombinant GSK-3 supplied by New England Biolabs is Rabbit GSK3-beta derived from a Rabbit skeletal muscle cDNA library. (see pmid: 7514173) taira_junichi@med.kurume-u.ac.jp J. Biochem. (0021-924X) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. taira-2014-4 Phosphorylation of Grb14 BPS domain by GSK-3 correlates with complex forming of Grb14 and insulin receptor. in vitro In vitro protein kinase assay protein kinase assay ms participant Identification by mass spectrometry ms participant Taira J., Higashimoto Y. Diabetes - Interactions investigated in the context of Diabetes Kinase-Glo Luminescent Kinase Assay (Promega);quantitating the amount of ATP remaining in solution following a kinase reaction. The luminescent signal is correlated with the amount of ATP present and is inversely correlated with the amount of kinase activity. IR cDNA is supplied by Kazusa DNA Research Institute (pFN21AE3464). The ORF corresponds to a protein of 1370aa, corresponding to Uniprot ID P06213-2 . GRB14 cDNA, protein_id="BAG35971.1" has 99% homology to the sequence given in the reviewed UniProt entry ID= Q14449. Recombinant GSK-3 supplied by New England Biolabs is Rabbit GSK3-beta derived from a Rabbit skeletal muscle cDNA library. (see pmid: 7514173) taira_junichi@med.kurume-u.ac.jp J. Biochem. (0021-924X) 2014 Only protein-protein interactions imex curation Affinomics - Interactions curated for the Affinomics consortium. gsk3b_human Glycogen synthase kinase-3 beta GSK3B Serine/threonine-protein kinase GSK3B protein protein human Homo sapiens Human MSGRPRTTSFAESCKPVQQPSAFGSMKVSRDKDGSKVTTVVATPGQGPDRPQEVSYTDTKVIGNGSFGVVYQAKLCDSGELVAIKKVLQDKRFKNRELQIMRKLDHCNIVRLRYFFYSSGEKKDEVYLNLVLDYVPETVYRVARHYSRAKQTLPVIYVKLYMYQLFRSLAYIHSFGICHRDIKPQNLLLDPDTAVLKLCDFGSAKQLVRGEPNVSYICSRYYRAPELIFGATDYTSSIDVWSAGCVLAELLLGQPIFPGDSGVDQLVEIIKVLGTPTREQIREMNPNYTEFKFPQIKAHPWTKVFRPRTPPEAIALCSRLLEYTPTARLTPLEACAHSFFDELRDPNVKLPNGRDTPALFNFTTQELSSNPPLATILIPPHARIQAAASTPTNATAASDANTGDRGQTNNAASASASNST grb14_human Growth factor receptor-bound protein 14 GRB14 adapter protein GRB14 protein protein human Homo sapiens Human MTTSLQDGQSAASRAAARDSPLAAQVCGAAQGRGDAHDLAPAPWLHARALLPLPDGTRGCAADRRKKKDLDVPEMPSIPNPFPELCCSPFTSVLSADLFPKANSRKKQVIKVYSEDETSRALDVPSDITARDVCQLLILKNHYIDDHSWTLFEHLPHIGVERTIEDHELVIEVLSNWGIEEENKLYFRKNYAKYEFFKNPMYFFPEHMVSFATETNGEISPTQILQMFLSSSTYPEIHGFLHAKEQGKKSWKKIYFFLRRSGLYFSTKGTSKEPRHLQFFSEFGNSDIYVSLAGKKKHGAPTNYGFCFKPNKAGGPRDLKMLCAEEEQSRTCWVTAIRLLKYGMQLYQNYMHPYQGRSGCSSQSISPMRSISENSLVAMDFSGQKSRVIENPTEALSVAVEEGLAWRKKGCLRLGTHGSPTASSQSSATNMAIHRSQPWFHHKISRDEAQRLIIQQGLVDGVFLVRDSQSNPKTFVLSMSHGQKIKHFQIIPVEDDGEMFHTLDDGHTRFTDLIQLVEFYQLNKGVLPCKLKHYCARIAL p06213-2 Insulin receptor HIR-A INSR protein protein human Homo sapiens Human MATGGRRGAAAAPLLVAVAALLLGAAGHLYPGEVCPGMDIRNNLTRLHELENCSVIEGHLQILLMFKTRPEDFRDLSFPKLIMITDYLLLFRVYGLESLKDLFPNLTVIRGSRLFFNYALVIFEMVHLKELGLYNLMNITRGSVRIEKNNELCYLATIDWSRILDSVEDNYIVLNKDDNEECGDICPGTAKGKTNCPATVINGQFVERCWTHSHCQKVCPTICKSHGCTAEGLCCHSECLGNCSQPDDPTKCVACRNFYLDGRCVETCPPPYYHFQDWRCVNFSFCQDLHHKCKNSRRQGCHQYVIHNNKCIPECPSGYTMNSSNLLCTPCLGPCPKVCHLLEGEKTIDSVTSAQELRGCTVINGSLIINIRGGNNLAAELEANLGLIEEISGYLKIRRSYALVSLSFFRKLRLIRGETLEIGNYSFYALDNQNLRQLWDWSKHNLTITQGKLFFHYNPKLCLSEIHKMEEVSGTKGRQERNDIALKTNGDQASCENELLKFSYIRTSFDKILLRWEPYWPPDFRDLLGFMLFYKEAPYQNVTEFDGQDACGSNSWTVVDIDPPLRSNDPKSQNHPGWLMRGLKPWTQYAIFVKTLVTFSDERRTYGAKSDIIYVQTDATNPSVPLDPISVSNSSSQIILKWKPPSDPNGNITHYLVFWERQAEDSELFELDYCLKGLKLPSRTWSPPFESEDSQKHNQSEYEDSAGECCSCPKTDSQILKELEESSFRKTFEDYLHNVVFVPRPSRKRRSLGDVGNVTVAVPTVAAFPNTSSTSVPTSPEEHRPFEKVVNKESLVISGLRHFTGYRIELQACNQDTPEERCSVAAYVSARTMPEAKADDIVGPVTHEIFENNVVHLMWQEPKEPNGLIVLYEVSYRRYGDEELHLCVSRKHFALERGCRLRGLSPGNYSVRIRATSLAGNGSWTEPTYFYVTDYLDVPSNIAKIIIGPLIFVFLFSVVIGSIYLFLRKRQPDGPLGPLYASSNPEYLSASDVFPCSVYVPDEWEVSREKITLLRELGQGSFGMVYEGNARDIIKGEAETRVAVKTVNESASLRERIEFLNEASVMKGFTCHHVVRLLGVVSKGQPTLVVMELMAHGDLKSYLRSLRPEAENNPGRPPPTLQEMIQMAAEIADGMAYLNAKKFVHRDLAARNCMVAHDFTVKIGDFGMTRDIYETDYYRKGGKGLLPVRWMAPESLKDGVFTTSSDMWSFGVVLWEITSLAEQPYQGLSNEQVLKFVMDGGYLDQPDNCPERVTDLMRMCWQFNPKMRPTFLEIVNLLKDDLHPSFPEVSFFHSEENKAPESEELEMEFEDMENVPLDRSSHCQREEAGGRDGGSSLGFKRSYEEHIPYTHMNGGKKNGRILTLPRSNPS csk21_human Casein kinase II subunit alpha CSNK2A1 CK2A1 protein protein human Homo sapiens Human MSGPVPSRARVYTDVNTHRPREYWDYESHVVEWGNQDDYQLVRKLGRGKYSEVFEAINITNNEKVVVKILKPVKKKKIKREIKILENLRGGPNIITLADIVKDPVSRTPALVFEHVNNTDFKQLYQTLTDYDIRFYMYEILKALDYCHSMGIMHRDVKPHNVMIDHEHRKLRLIDWGLAEFYHPGQEYNVRVASRYFKGPELLVDYQMYDYSLDMWSLGCMLASMIFRKEPFFHGHDNYDQLVRIAKVLGTEDLYDYIDKYNIELDPRFNDILGRHSRKRWERFVHSENQHLVSPEALDFLDKLLRYDHQSRLTAREAMEHPYFYTVVKDQARMGSSSMPGGSTPVSSANMMSGISSVPTPSPLGPLAGSPVIAAANPLGMPVPAAAGAQQ csk2b_human Casein kinase II subunit beta Phosvitin Protein G5a CSNK2B CK2N G5A protein protein human Homo sapiens Human MSSSEEVSWISWFCGLRGNEFFCEVDEDYIQDKFNLTGLNEQVPHYRQALDMILDLEPDEELEDNPNQSDLIEQAAEMLYGLIHARYILTNRGIAQMLEKYQQGDFGYCPRVYCENQPMLPIGLSDIPGEAMVKLYCPKCMDVYTPKSSRHHHTDGAYFGTGFPHMLFMVHPEYRPKRPANQFVPRLYGFKIHPMAYQLQLQAASNFKSPVKTIR rabbit_gsk3_beta RABBIT GSK3-BETA protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit grb14-gsk3b 1600238 n/a 1600242 unspecified role unspecified role neutral component neutral component n/a 1600243 unspecified role unspecified role neutral component neutral component c_terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false 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 false proximity proximity close-contact colocalization neighbourhood interaction false false false 2A A robust signal was obtained using transfected Human Grb14 and GSK-3beta; however endogenous Grb14, GSK-3alpha and GSK-3beta were also detected. grb14-insr-1 1600239 n/a 1600244 unspecified role unspecified role prey prey n/a 1600243 unspecified role unspecified role bait bait Q14449:p.[Ser358Ala;Ser362Ala;Ser366Ala] mutation increasing strength mutation increasing interaction strength mutation increasing strength 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 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 false c_terminus v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false 3/4/6 Insulin GSK-3 inhibitor IX GSK-3 knockdown insr-csnk2a1 1600240 n/a 1600245 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1600243 enzyme target enzyme target substrate substrate neutral component neutral component bps_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1600246 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 5A Recombinant Rabbit GSK-3beta Q14449 O-phospho-L-serine grb14-rabbit 1600241 n/a 1600247 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1600243 enzyme target enzyme target substrate substrate neutral component neutral component required_phosphoser_370 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain false bps_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false 5A Q14449 opser After GSK-3 incubation using the Ser370 phosphorylated peptide Grb14(355–375) signals corresponding to bi-, tri- and tetra-phosphorylated peptides were found; using the non-phosphorylated peptide a signal corresponded to a non-phosphorylated peptide was found, with a marginal signal indicating mono-phosphorylated peptides.