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/ mcnulty-2011-1 MAPK scaffold IQGAP1 binds the EGF receptor and modulates its activation. 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 McNulty D.E., Li Z., White C.D., Sacks D.B., Annan R.S. Cancer - Interactions investigated in the context of cancer Species of constructs/proteins used was inferred from sequence/length/mutation location info. (For EGFR T790M mutation) GST-IQGAP1 origin was found from Hart et al, 1996, EMBO Journal, pp.2997-3005. (D29640). sacksdb@mail.nih.gov, roland.s.annan@gsk.com J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation mcnulty-2011-2 MAPK scaffold IQGAP1 binds the EGF receptor and modulates its activation. human-a_431 Homo sapiens epidermoid carcinoma a_431 Human epidermoid carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot McNulty D.E., Li Z., White C.D., Sacks D.B., Annan R.S. Cancer - Interactions investigated in the context of cancer Species of constructs/proteins used was inferred from sequence/length/mutation location info. (For EGFR T790M mutation) GST-IQGAP1 origin was found from Hart et al, 1996, EMBO Journal, pp.2997-3005. (D29640). sacksdb@mail.nih.gov, roland.s.annan@gsk.com J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation mcnulty-2011-3 MAPK scaffold IQGAP1 binds the EGF receptor and modulates its activation. human-a_431 Homo sapiens epidermoid carcinoma a_431 Human epidermoid carcinoma confocal microscopy confocal microscopy immunostaining immunostaining McNulty D.E., Li Z., White C.D., Sacks D.B., Annan R.S. Cancer - Interactions investigated in the context of cancer Species of constructs/proteins used was inferred from sequence/length/mutation location info. (For EGFR T790M mutation) GST-IQGAP1 origin was found from Hart et al, 1996, EMBO Journal, pp.2997-3005. (D29640). sacksdb@mail.nih.gov, roland.s.annan@gsk.com J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation mcnulty-2011-4 MAPK scaffold IQGAP1 binds the EGF receptor and modulates its activation. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot McNulty D.E., Li Z., White C.D., Sacks D.B., Annan R.S. Cancer - Interactions investigated in the context of cancer Species of constructs/proteins used was inferred from sequence/length/mutation location info. (For EGFR T790M mutation) GST-IQGAP1 origin was found from Hart et al, 1996, EMBO Journal, pp.2997-3005. (D29640). sacksdb@mail.nih.gov, roland.s.annan@gsk.com J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation mcnulty-2011-5 MAPK scaffold IQGAP1 binds the EGF receptor and modulates its activation. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra McNulty D.E., Li Z., White C.D., Sacks D.B., Annan R.S. Cancer - Interactions investigated in the context of cancer The natural variant/mutation thr790met has been found in a lung cancer sample. It results in increased kinase activity and causes drug resistance by increasing the affinity for ATP. Species of constructs/proteins used was inferred from sequence/length/mutation location info. (For EGFR T790M mutation) GST-IQGAP1 origin was found from Hart et al, 1996, EMBO Journal, pp.2997-3005. (D29640). sacksdb@mail.nih.gov, roland.s.annan@gsk.com J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation iqga1_human Ras GTPase-activating-like protein IQGAP1 IQGAP1 KIAA0051 p195 protein protein human Homo sapiens Human MSAADEVDGLGVARPHYGSVLDNERLTAEEMDERRRQNVAYEYLCHLEEAKRWMEACLGEDLPPTTELEEGLRNGVYLAKLGNFFSPKVVSLKKIYDREQTRYKATGLHFRHTDNVIQWLNAMDEIGLPKIFYPETTDIYDRKNMPRCIYCIHALSLYLFKLGLAPQIQDLYGKVDFTEEEINNMKTELEKYGIQMPAFSKIGGILANELSVDEAALHAAVIAINEAIDRRIPADTFAALKNPNAMLVNLEEPLASTYQDILYQAKQDKMTNAKNRTENSERERDVYEELLTQAEIQGNINKVNTFSALANIDLALEQGDALALFRALQSPALGLRGLQQQNSDWYLKQLLSDKQQKRQSGQTDPLQKEELQSGVDAANSAAQQYQRRLAAVALINAAIQKGVAEKTVLELMNPEAQLPQVYPFAADLYQKELATLQRQSPEHNLTHPELSVAVEMLSSVALINRALESGDVNTVWKQLSSSVTGLTNIEEENCQRYLDELMKLKAQAHAENNEFITWNDIQACVDHVNLVVQEEHERILAIGLINEALDEGDAQKTLQALQIPAAKLEGVLAEVAQHYQDTLIRAKREKAQEIQDESAVLWLDEIQGGIWQSNKDTQEAQKFALGIFAINEAVESGDVGKTLSALRSPDVGLYGVIPECGETYHSDLAEAKKKKLAVGDNNSKWVKHWVKGGYYYYHNLETQEGGWDEPPNFVQNSMQLSREEIQSSISGVTAAYNREQLWLANEGLITRLQARCRGYLVRQEFRSRMNFLKKQIPAITCIQSQWRGYKQKKAYQDRLAYLRSHKDEVVKIQSLARMHQARKRYRDRLQYFRDHINDIIKIQAFIRANKARDDYKTLINAEDPPMVVVRKFVHLLDQSDQDFQEELDLMKMREEVITLIRSNQQLENDLNLMDIKIGLLVKNKITLQDVVSHSKKLTKKNKEQLSDMMMINKQKGGLKALSKEKREKLEAYQHLFYLLQTNPTYLAKLIFQMPQNKSTKFMDSVIFTLYNYASNQREEYLLLRLFKTALQEEIKSKVDQIQEIVTGNPTVIKMVVSFNRGARGQNALRQILAPVVKEIMDDKSLNIKTDPVDIYKSWVNQMESQTGEASKLPYDVTPEQALAHEEVKTRLDSSIRNMRAVTDKFLSAIVSSVDKIPYGMRFIAKVLKDSLHEKFPDAGEDELLKIIGNLLYYRYMNPAIVAPDAFDIIDLSAGGQLTTDQRRNLGSIAKMLQHAASNKMFLGDNAHLSIINEYLSQSYQKFRRFFQTACDVPELQDKFNVDEYSDLVTLTKPVIYISIGEIINTHTLLLDHQDAIAPEHNDPIHELLDDLGEVPTIESLIGESSGNLNDPNKEALAKTEVSLTLTNKFDVPGDENAEMDARTILLNTKRLIVDVIRFQPGETLTEILETPATSEQEAEHQRAMQRRAIRDAKTPDKMKKSKSVKEDSNLTLQEKKEKIQTGLKKLTELGTVDPKNKYQELINDIARDIRNQRRYRQRRKAELVKLQQTYAALNSKATFYGEQVDYYKSYIKTCLDNLASKGKVSKKPREMKGKKSKKISLKYTAARLHEKGVLLEIEDLQVNQFKNVIFEISPTEEVGDFEVKAKFMGVQMETFMLHYQDLLQLQYEGVAVMKLFDRAKVNVNLLIFLLNKKFYGK egfr_chlae EGFR Green Monkey protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey egfr_human Epidermal growth factor receptor ERBB1 EGFR Receptor tyrosine-protein kinase erbB-1 Proto-oncogene c-ErbB-1 ERBB HER1 protein protein human Homo sapiens Human MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFEDHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENLQIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQWRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGRCRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNPEGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGIGIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTPSGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSPKANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQYLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHAEGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKGERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHLPSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVACIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQNPVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQISLDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA iqgap1-egfr 1268480 n/a 1268485 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1268486 unspecified role unspecified role prey prey physical association physical association false false false Fig 1A iqgap1-egfr-1 1268481 n/a 1268487 unspecified role unspecified role prey prey n/a 1268485 unspecified role unspecified role bait bait physical association physical association false false false Fig 1B, 1C. Ca2+ ions egfr-iqgap1 1268482 n/a 1268487 unspecified role unspecified role neutral component neutral component n/a 1268485 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig 2 EGF induces translocation of both IQGAP1 and EGFR to the periphery of cells, where the proteins colocalize in areas of ruffling consistent with lamellipodial structures. iqgap1-egfr-2 1268483 n/a 1268487 unspecified role unspecified role prey prey protein_kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 n/a 1268485 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig 3 calmodulin, and calcium free calmodulin disrupt the interaction. egfr-iqgap1-1 1268484 n/a 1268487 unspecified role unspecified role bait bait intracellular_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 P00533:p.Thr790Met mutation mutation certain certain sequence position certain certain certain sequence position certain 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 n/a 1268485 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 1q_domains required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false in vitro In vitro 1268510 1268505 direct interaction direct interaction false false false Fig 4