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/ hara-2002-2 Raptor, a binding partner of target of rapamycin (TOR), mediates TOR action. 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 anti tag western anti tag western blot anti tag western Hara K., Maruki Y., Long X., Yoshino K., Oshiro N., Hidayat S., Tokunaga C., Avruch J., Yonezawa K. Apoptosis - Interactions involving proteins with a function related to apoptosis First, mTOR and its potential binding partners were precipitated with 30% (NH4)2SO4. Then, the authors did a ion column chromatography. The immunopurified proteins were further purified by immunoprecipitation and identified by peptide mass fingerprinting and tandem mass spectrometry to identify the participants. yonezawa@kobe-u.ac.jp,harak@kobe-u.ac.jp specific monoclonal antibody against mTOR for the immunoprecipitation Cell (0092-8674) 2002 Only protein-protein interactions imex curation hara-2002-4 Raptor, a binding partner of target of rapamycin (TOR), mediates TOR action. in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Hara K., Maruki Y., Long X., Yoshino K., Oshiro N., Hidayat S., Tokunaga C., Avruch J., Yonezawa K. Apoptosis - Interactions involving proteins with a function related to apoptosis First, mTOR and its potential binding partners were precipitated with 30% (NH4)2SO4. Then, the authors did a ion column chromatography. The immunopurified proteins were further purified by immunoprecipitation and identified by peptide mass fingerprinting and tandem mass spectrometry to identify the participants. yonezawa@kobe-u.ac.jp,harak@kobe-u.ac.jp specific monoclonal antibody against mTOR for the immunoprecipitation Cell (0092-8674) 2002 Only protein-protein interactions imex curation hara-2002-5 Raptor, a binding partner of target of rapamycin (TOR), mediates TOR action. 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. pull down pull down anti tag western anti tag western blot anti tag western Hara K., Maruki Y., Long X., Yoshino K., Oshiro N., Hidayat S., Tokunaga C., Avruch J., Yonezawa K. Apoptosis - Interactions involving proteins with a function related to apoptosis First, mTOR and its potential binding partners were precipitated with 30% (NH4)2SO4. Then, the authors did a ion column chromatography. The immunopurified proteins were further purified by immunoprecipitation and identified by peptide mass fingerprinting and tandem mass spectrometry to identify the participants. yonezawa@kobe-u.ac.jp,harak@kobe-u.ac.jp specific monoclonal antibody against mTOR for the immunoprecipitation Cell (0092-8674) 2002 Only protein-protein interactions imex curation hara-2002-3 Raptor, a binding partner of target of rapamycin (TOR), mediates TOR action. 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Hara K., Maruki Y., Long X., Yoshino K., Oshiro N., Hidayat S., Tokunaga C., Avruch J., Yonezawa K. Apoptosis - Interactions involving proteins with a function related to apoptosis First, mTOR and its potential binding partners were precipitated with 30% (NH4)2SO4. Then, the authors did a ion column chromatography. The immunopurified proteins were further purified by immunoprecipitation and identified by peptide mass fingerprinting and tandem mass spectrometry to identify the participants. yonezawa@kobe-u.ac.jp,harak@kobe-u.ac.jp specific monoclonal antibody against mTOR for the immunoprecipitation Cell (0092-8674) 2002 Only protein-protein interactions imex curation hara-2002-1 Raptor, a binding partner of target of rapamycin (TOR), mediates TOR action. 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 bait coip anti bait coimmunoprecipitation anti bait coip sequence tag sequence tag identification sequence tag Hara K., Maruki Y., Long X., Yoshino K., Oshiro N., Hidayat S., Tokunaga C., Avruch J., Yonezawa K. Apoptosis - Interactions involving proteins with a function related to apoptosis First, mTOR and its potential binding partners were precipitated with 30% (NH4)2SO4. Then, the authors did a ion column chromatography. The immunopurified proteins were further purified by immunoprecipitation and identified by peptide mass fingerprinting and tandem mass spectrometry to identify the participants. yonezawa@kobe-u.ac.jp,harak@kobe-u.ac.jp specific monoclonal antibody against mTOR for the immunoprecipitation Cell (0092-8674) 2002 Only protein-protein interactions imex curation rptor_human Regulatory-associated protein of mTOR p150 target of rapamycin (TOR)-scaffold protein RPTOR KIAA1303 RAPTOR protein protein human Homo sapiens Human MESEMLQSPLLGLGEEDEADLTDWNLPLAFMKKRHCEKIEGSKSLAQSWRMKDRMKTVSVALVLCLNVGVDPPDVVKTTPCARLECWIDPLSMGPQKALETIGANLQKQYENWQPRARYKQSLDPTVDEVKKLCTSLRRNAKEERVLFHYNGHGVPRPTVNGEVWVFNKNYTQYIPLSIYDLQTWMGSPSIFVYDCSNAGLIVKSFKQFALQREQELEVAAINPNHPLAQMPLPPSMKNCIQLAACEATELLPMIPDLPADLFTSCLTTPIKIALRWFCMQKCVSLVPGVTLDLIEKIPGRLNDRRTPLGELNWIFTAITDTIAWNVLPRDLFQKLFRQDLLVASLFRNFLLAERIMRSYNCTPVSSPRLPPTYMHAMWQAWDLAVDICLSQLPTIIEEGTAFRHSPFFAEQLTAFQVWLTMGVENRNPPEQLPIVLQVLLSQVHRLRALDLLGRFLDLGPWAVSLALSVGIFPYVLKLLQSSARELRPLLVFIWAKILAVDSSCQADLVKDNGHKYFLSVLADPYMPAEHRTMTAFILAVIVNSYHTGQEACLQGNLIAICLEQLNDPHPLLRQWVAICLGRIWQNFDSARWCGVRDSAHEKLYSLLSDPIPEVRCAAVFALGTFVGNSAERTDHSTTIDHNVAMMLAQLVSDGSPMVRKELVVALSHLVVQYESNFCTVALQFIEEEKNYALPSPATTEGGSLTPVRDSPCTPRLRSVSSYGNIRAVATARSLNKSLQNLSLTEESGGAVAFSPGNLSTSSSASSTLGSPENEEHILSFETIDKMRRASSYSSLNSLIGVSFNSVYTQIWRVLLHLAADPYPEVSDVAMKVLNSIAYKATVNARPQRVLDTSSLTQSAPASPTNKGVHIHQAGGSPPASSTSSSSLTNDVAKQPVSRDLPSGRPGTTGPAGAQYTPHSHQFPRTRKMFDKGPEQTADDADDAAGHKSFISATVQTGFCDWSARYFAQPVMKIPEEHDLESQIRKEREWRFLRNSRVRRQAQQVIQKGITRLDDQIFLNRNPGVPSVVKFHPFTPCIAVADKDSICFWDWEKGEKLDYFHNGNPRYTRVTAMEYLNGQDCSLLLTATDDGAIRVWKNFADLEKNPEMVTAWQGLSDMLPTTRGAGMVVDWEQETGLLMSSGDVRIVRIWDTDREMKVQDIPTGADSCVTSLSCDSHRSLIVAGLGDGSIRVYDRRMALSECRVMTYREHTAWVVKASLQKRPDGHIVSVSVNGDVRIFDPRMPESVNVLQIVKGLTALDIHPQADLIACGSVNQFTAIYNSSGELINNIKYYDGFMGQRVGAISCLAFHPHWPHLAVGSNDYYISVYSVEKRVR mtor_mouse Serine/threonine-protein kinase mTOR FK506-binding protein 12-rapamycin complex-associated protein 1 Rapamycin target protein 1 Mtor Mammalian target of rapamycin Frap1 Frap FKBP12-rapamycin complex-associated protein Mechanistic target of rapamycin protein protein mouse Mus musculus Mouse MLGTGPAVATASAATSSNVSVLQQFASGLKSRNEETRAKAAKELQHYVTMELREMSQEESTRFYDQLNHHIFELVSSSDANERKGGILAIASLIGVEGGNSTRIGRFANYLRNLLPSSDPVVMEMASKAIGRLAMAGDTFTAEYVEFEVKRALEWLGADRNEGRRHAAVLVLRELAISVPTFFFQQVQPFFDNIFVAVWDPKQAIREGAVAALRACLILTTQREPKEMQKPQWYRHTFEEAEKGFDETLAKEKGMNRDDRIHGALLILNELVRISSMEGERLREEMEEITQQQLVHDKYCKDLMGFGTKPRHITPFTSFQAVQPQQPNALVGLLGYSSPQGLMGFGTSPSPAKSTLVESRCCRDLMEEKFDQVCQWVLKCRSSKNSLIQMTILNLLPRLAAFRPSAFTDTQYLQDTMNHVLSCVKKEKERTAAFQALGLLSVAVRSEFKVYLPRVLDIIRAALPPKDFAHKRQKTVQVDATVFTCISMLARAMGPGIQQDIKELLEPMLAVGLSPALTAVLYDLSRQIPQLKKDIQDGLLKMLSLVLMHKPLRHPGMPKGLAHQLASPGLTTLPEASDVASITLALRTLGSFEFEGHSLTQFVRHCADHFLNSEHKEIRMEAARTCSRLLTPSIHLISGHAHVVSQTAVQVVADVLSKLLVVGITDPDPDIRYCVLASLDERFDAHLAQAENLQALFVALNDQVFEIRELAICTVGRLSSMNPAFVMPFLRKMLIQILTELEHSGIGRIKEQSARMLGHLVSNAPRLIRPYMEPILKALILKLKDPDPDPNPGVINNVLATIGELAQVSGLEMRKWVDELFIIIMDMLQDSSLLAKRQVALWTLGQLVASTGYVVEPYRKYPTLLEVLLNFLKTEQNQGTRREAIRVLGLLGALDPYKHKVNIGMIDQSRDASAVSLSESKSSQDSSDYSTSEMLVNMGNLPLDEFYPAVSMVALMRIFRDQSLSHHHTMVVQAITFIFKSLGLKCVQFLPQVMPTFLNVIRVCDGAIREFLFQQLGMLVSFVKSHIRPYMDEIVTLMREFWVMNTSIQSTIILLIEQIVVALGGEFKLYLPQLIPHMLRVFMHDNSQGRIVSIKLLAAIQLFGANLDDYLHLLLPPIVKLFDAPEVPLPSRKAALETVDRLTESLDFTDYASRIIHPIVRTLDQSPELRSTAMDTLSSLVFQLGKKYQIFIPMVNKVLVRHRINHQRYDVLICRIVKGYTLADEEEDPLIYQHRMLRSSQGDALASGPVETGPMKKLHVSTINLQKAWGAARRVSKDDWLEWLRRLSLELLKDSSSPSLRSCWALAQAYNPMARDLFNAAFVSCWSELNEDQQDELIRSIELALTSQDIAEVTQTLLNLAEFMEHSDKGPLPLRDDNGIVLLGERAAKCRAYAKALHYKELEFQKGPTPAILESLISINNKLQQPEAASGVLEYAMKHFGELEIQATWYEKLHEWEDALVAYDKKMDTNKEDPELMLGRMRCLEALGEWGQLHQQCCEKWTLVNDETQAKMARMAAAAAWGLGQWDSMEEYTCMIPRDTHDGAFYRAVLALHQDLFSLAQQCIDKARDLLDAELTAMAGESYSRAYGAMVSCHMLSELEEVIQYKLVPERREIIRQIWWERLQGCQRIVEDWQKILMVRSLVVSPHEDMRTWLKYASLCGKSGRLALAHKTLVLLLGVDPSRQLDHPLPTAHPQVTYAYMKNMWKSARKIDAFQHMQHFVQTMQQQAQHAIATEDQQHKQELHKLMARCFLKLGEWQLNLQGINESTIPKVLQYYSAATEHDRSWYKAWHAWAVMNFEAVLHYKHQNQARDEKKKLRHASGANITNATTAATTAASAAAATSTEGSNSESEAESNENSPTPSPLQKKVTEDLSKTLLLYTVPAVQGFFRSISLSRGNNLQDTLRVLTLWFDYGHWPDVNEALVEGVKAIQIDTWLQVIPQLIARIDTPRPLVGRLIHQLLTDIGRYHPQALIYPLTVASKSTTTARHNAANKILKNMCEHSNTLVQQAMMVSEELIRVAILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQLPQLTSLELQYVSPKLLMCRDLELAVPGTYDPNQPIIRIQSIAPSLQVITSKQRPRKLTLMGSNGHEFVFLLKGHEDLRQDERVMQLFGLVNTLLANDPTSLRKNLSIQRYAVIPLSTNSGLIGWVPHCDTLHALIRDYREKKKILLNIEHRIMLRMAPDYDHLTLMQKVEVFEHAVNNTAGDDLAKLLWLKSPSSEVWFDRRTNYTRSLAVMSMVGYILGLGDRHPSNLMLDRLSGKILHIDFGDCFEVAMTREKFPEKIPFRLTRMLTNAMEVTGLDGNYRTTCHTVMEVLREHKDSVMAVLEAFVYDPLLNWRLMDTNTKGNKRSRTRTDSYSAGQSVEILDGVELGEPAHKKAGTTVPESIHSFIGDGLVKPEALNKKAIQIINRVRDKLTGRDFSHDDTLDVPTQVELLIKQATSHENLCQCYIGWCPFW 4ebp1_human Eukaryotic translation initiation factor 4E-binding protein 1 Phosphorylated heat- and acid-stable protein regulated by insulin 1 EIF4EBP1 protein protein human Homo sapiens Human MSGGSSCSQTPSRAIPATRRVVLGDGVQLPPGDYSTTPGGTLFSTTPGGTRIIYDRKFLMECRNSPVTKTPPRDLPTIPGVTSPSSDEPPMEASQSHLRNSPEDKRAGGEESQFEMDI ks6b1_rat Ribosomal protein S6 kinase beta-1 Rps6kb1 Ribosomal protein S6 kinase I 70 kDa ribosomal protein S6 kinase 1 p70 ribosomal S6 kinase alpha protein protein rat Rattus norvegicus (Rat) Rat MRRRRRRDGFYPAPDFRHREAEDMAGVFDIDLDQPEDAGSEDELEEGGQLNESMDHGGVGPYELGMEHCEKFEISETSVNRGPEKIRPECFELLRVLGKGGYGKVFQVRKVTGANTGKIFAMKVLKKAMIVRNAKDTAHTKAERNILEEVKHPFIVDLIYAFQTGGKLYLILEYLSGGELFMQLEREGIFMEDTACFYLAEISMALGHLHQKGIIYRDLKPENIMLNHQGHVKLTDFGLCKESIHDGTVTHTFCGTIEYMAPEILMRSGHNRAVDWWSLGALMYDMLTGAPPFTGENRKKTIDKILKCKLNLPPYLTQEARDLLKKLLKRNAASRLGAGPGDAGEVQAHPFFRHINWEELLARKVEPPFKPLLQSEEDVSQFDSKFTRQTPVDSPDDSTLSESANQVFLGFTYVAPSVLESVKEKFSFEPKIRSPRRFIGSPRTPVSPVKFSPGDFWGRGASASTANPQTPVEYPMETSGIEQMDVTTSGEASAPLPIRQPNSGPYKKQAFPMISKRPEHLRMNL mtor_human Serine/threonine-protein kinase mTOR MTOR FRAP FRAP2 FK506-binding protein 12-rapamycin complex-associated protein 1 Rapamycin target protein 1 FRAP1 Mammalian target of rapamycin FKBP12-rapamycin complex-associated protein Mechanistic target of rapamycin RAFT1 RAPT1 Rapamycin and FKBP12 target 1 protein protein human Homo sapiens Human MLGTGPAAATTAATTSSNVSVLQQFASGLKSRNEETRAKAAKELQHYVTMELREMSQEESTRFYDQLNHHIFELVSSSDANERKGGILAIASLIGVEGGNATRIGRFANYLRNLLPSNDPVVMEMASKAIGRLAMAGDTFTAEYVEFEVKRALEWLGADRNEGRRHAAVLVLRELAISVPTFFFQQVQPFFDNIFVAVWDPKQAIREGAVAALRACLILTTQREPKEMQKPQWYRHTFEEAEKGFDETLAKEKGMNRDDRIHGALLILNELVRISSMEGERLREEMEEITQQQLVHDKYCKDLMGFGTKPRHITPFTSFQAVQPQQSNALVGLLGYSSHQGLMGFGTSPSPAKSTLVESRCCRDLMEEKFDQVCQWVLKCRNSKNSLIQMTILNLLPRLAAFRPSAFTDTQYLQDTMNHVLSCVKKEKERTAAFQALGLLSVAVRSEFKVYLPRVLDIIRAALPPKDFAHKRQKAMQVDATVFTCISMLARAMGPGIQQDIKELLEPMLAVGLSPALTAVLYDLSRQIPQLKKDIQDGLLKMLSLVLMHKPLRHPGMPKGLAHQLASPGLTTLPEASDVGSITLALRTLGSFEFEGHSLTQFVRHCADHFLNSEHKEIRMEAARTCSRLLTPSIHLISGHAHVVSQTAVQVVADVLSKLLVVGITDPDPDIRYCVLASLDERFDAHLAQAENLQALFVALNDQVFEIRELAICTVGRLSSMNPAFVMPFLRKMLIQILTELEHSGIGRIKEQSARMLGHLVSNAPRLIRPYMEPILKALILKLKDPDPDPNPGVINNVLATIGELAQVSGLEMRKWVDELFIIIMDMLQDSSLLAKRQVALWTLGQLVASTGYVVEPYRKYPTLLEVLLNFLKTEQNQGTRREAIRVLGLLGALDPYKHKVNIGMIDQSRDASAVSLSESKSSQDSSDYSTSEMLVNMGNLPLDEFYPAVSMVALMRIFRDQSLSHHHTMVVQAITFIFKSLGLKCVQFLPQVMPTFLNVIRVCDGAIREFLFQQLGMLVSFVKSHIRPYMDEIVTLMREFWVMNTSIQSTIILLIEQIVVALGGEFKLYLPQLIPHMLRVFMHDNSPGRIVSIKLLAAIQLFGANLDDYLHLLLPPIVKLFDAPEAPLPSRKAALETVDRLTESLDFTDYASRIIHPIVRTLDQSPELRSTAMDTLSSLVFQLGKKYQIFIPMVNKVLVRHRINHQRYDVLICRIVKGYTLADEEEDPLIYQHRMLRSGQGDALASGPVETGPMKKLHVSTINLQKAWGAARRVSKDDWLEWLRRLSLELLKDSSSPSLRSCWALAQAYNPMARDLFNAAFVSCWSELNEDQQDELIRSIELALTSQDIAEVTQTLLNLAEFMEHSDKGPLPLRDDNGIVLLGERAAKCRAYAKALHYKELEFQKGPTPAILESLISINNKLQQPEAAAGVLEYAMKHFGELEIQATWYEKLHEWEDALVAYDKKMDTNKDDPELMLGRMRCLEALGEWGQLHQQCCEKWTLVNDETQAKMARMAAAAAWGLGQWDSMEEYTCMIPRDTHDGAFYRAVLALHQDLFSLAQQCIDKARDLLDAELTAMAGESYSRAYGAMVSCHMLSELEEVIQYKLVPERREIIRQIWWERLQGCQRIVEDWQKILMVRSLVVSPHEDMRTWLKYASLCGKSGRLALAHKTLVLLLGVDPSRQLDHPLPTVHPQVTYAYMKNMWKSARKIDAFQHMQHFVQTMQQQAQHAIATEDQQHKQELHKLMARCFLKLGEWQLNLQGINESTIPKVLQYYSAATEHDRSWYKAWHAWAVMNFEAVLHYKHQNQARDEKKKLRHASGANITNATTAATTAATATTTASTEGSNSESEAESTENSPTPSPLQKKVTEDLSKTLLMYTVPAVQGFFRSISLSRGNNLQDTLRVLTLWFDYGHWPDVNEALVEGVKAIQIDTWLQVIPQLIARIDTPRPLVGRLIHQLLTDIGRYHPQALIYPLTVASKSTTTARHNAANKILKNMCEHSNTLVQQAMMVSEELIRVAILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQLPQLTSLELQYVSPKLLMCRDLELAVPGTYDPNQPIIRIQSIAPSLQVITSKQRPRKLTLMGSNGHEFVFLLKGHEDLRQDERVMQLFGLVNTLLANDPTSLRKNLSIQRYAVIPLSTNSGLIGWVPHCDTLHALIRDYREKKKILLNIEHRIMLRMAPDYDHLTLMQKVEVFEHAVNNTAGDDLAKLLWLKSPSSEVWFDRRTNYTRSLAVMSMVGYILGLGDRHPSNLMLDRLSGKILHIDFGDCFEVAMTREKFPEKIPFRLTRMLTNAMEVTGLDGNYRITCHTVMEVLREHKDSVMAVLEAFVYDPLLNWRLMDTNTKGNKRSRTRTDSYSAGQSVEILDGVELGEPAHKKTGTTVPESIHSFIGDGLVKPEALNKKAIQIINRVRDKLTGRDFSHDDTLDVPTQVELLIKQATSHENLCQCYIGWCPFW frap1-rptor-2 physical association of FRAP1 with RPTOR 906880 n/a 906885 unspecified role unspecified role prey prey region flag tagged 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 false n/a 906886 unspecified role unspecified role bait bait n-terminal HA tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 2. A and Fig. 3. A Q8N122;Q9JLN9 transfection the interaction is abolished if the immunoprecipitates are washed with a wash buffer containing 1% NP-40. rptor-frap1-1 physical association of RPTOR with FRAP1 906880 n/a 906886 unspecified role unspecified role prey prey n-terminal HA tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 906885 unspecified role unspecified role bait bait region flag tagged 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 false physical association physical association false false false Fig. 2. A Q8N122;Q9JLN9 transfection the interaction is abolished if the immunoprecipitates are washed with a wash buffer containing 1% NP-40. rptor-eif4ebp1-1 association of RPTOR with EIF4EBP1 and FRAP1 906880 n/a 906885 unspecified role unspecified role bait bait region flag tagged 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 false n/a 906887 unspecified role unspecified role prey prey region GST tagged 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 n/a 906886 unspecified role unspecified role prey prey n-terminal HA tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Fig. 4. B Q8N122;Q9JLN9;B2R502 transfection rptor-rps6kb1-1 association of RPTOR with Rps6kb1 and FRAP1 906880 n/a 906886 unspecified role unspecified role prey prey region HA tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 906885 unspecified role unspecified role bait bait region flag tagged 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 false n/a 906888 unspecified role unspecified role prey prey region GST tagged 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 association association false false false Fig. 4. B Q8N122;Q9JLN9;P67999 transfection rptor-eif4ebp1-2 physical association of RPTOR with EIF4EBP1 906880 n/a 906887 unspecified role unspecified role prey prey Q13541:p.[Thr37Glu;Thr46Glu;Ser65Glu;Thr70Glu;Ser83Glu] thr37glu_thr46glu_ser65glu_thr70glu_ser83glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region flag tagged 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 false Q13541:p.[Thr37Ala;Thr46Ala;Ser65Ala;Thr70Ala;Ser83Ala] thr37ala_thr46ala_ser65ala_thr70ala_ser83ala mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n/a 906885 unspecified role unspecified role bait bait region myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 5. C Q8N122;B2R502 transfection frap1-eif4ebp1-1 Phosphorylation of EIF4EBP1 by FRAP1 906881 n/a 906886 enzyme enzyme neutral component neutral component region HA tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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. n/a 906887 enzyme target enzyme target substrate substrate neutral component neutral component region GST tagged 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 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. phosphorylation phosphorylation reaction phosphorylation false false false Fig. 3. A and B Q9JLN9;B2R502 transfection Rapamycin decreases the phosphorylation of EIF4EBP1 by FRAP1 RPTOR (Q8N122) increases the kinase activity of FRAP1 EIF4EBP1 is phosphorylated by Frap1. frap1-rps6kb1-1 Phosphorylation of Rps6kb1 by FRAP1 906881 n/a 906888 enzyme target enzyme target substrate substrate neutral component neutral component range GST tagged 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 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. n/a 906886 enzyme enzyme neutral component neutral component n-terminal HA tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q9JLN9:p.Asn2343Lys Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false 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. phosphorylation phosphorylation reaction phosphorylation false false false Fig. 3. C P42345;P67999 transfection RPTOR (Q8N122) increases the kinase activity of FRAP1 specific monoclonal antibody against FRAP1 has been used to immunoprecipitate FRAP1 Rps6kb1 is phosphorylated by Frap1. The required amino acid for the kinase activity of Frap1 is asn 2343. frap1-eif4ebp1-2 Phosphorylation of EIF4EBP1 by FRAP1 906881 n/a 906889 enzyme enzyme neutral component neutral component 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. n/a 906887 enzyme target enzyme target substrate substrate neutral component neutral component region GST tagged 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 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. phosphorylation phosphorylation reaction phosphorylation false false false Fig. 2. B B2R502 transfection RPTOR (Q8N122) increases the kinase activity of FRAP1 specific monoclonal antibody against FRAP1 has been used to immunoprecipitate FRAP1 EIF4EBP1 is phosphorylated by FRAP1 frap1-eif4ebp1-3 Phosphorylation of EIF4EBP1 by FRAP1 906881 n/a 906887 enzyme target enzyme target substrate substrate neutral component neutral component region GST tagged 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 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. n/a 906889 enzyme enzyme neutral component neutral component human-hela_s3 Homo sapiens HeLa-S3 cervix carcinoma cell line hela_s3 HeLa-S3 cervix carcinoma cell line Human cervix carcinoma cell line is a subclone of its parent HELA (DSM ACC 57) derived in 1955 confirmed as human with IEF of AST, MDH phosphorylation phosphorylation reaction phosphorylation false false false Fig. 6. A B2R502 transfection RPTOR (Q8N122) increases the kinase activity of FRAP1 specific monoclonal antibody against FRAP1 has been used to immunoprecipitate FRAP1 The mTOR kinase activity toward GST-4EBP1 were severely reduced in cells transfected with siRNA 47-67 duplex (bp 47 to 67 of Raptor) but moderately reduced with siRNA 127-147 (bp 127 to 147 of raptor) Phosphorylation of EIF4EP1 eif4ebp1-rptor-1 physical association of EIF4EBP1 with RPTOR 906882 n/a 906887 unspecified role unspecified role bait bait region GST tagged 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 n/a 906885 unspecified role unspecified role prey prey n-terminal myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 4. A Q8N122;B2R502 transfection rps6kb1-rptor-1 physical association of Rps6kb1 with RPTOR 906882 n/a 906885 unspecified role unspecified role prey prey n-terminal myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 906888 unspecified role unspecified role bait bait region GST tagged 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 physical association physical association false false false Fig. 4. A and C Q8N122;P67999 transfection eif4ebp1-rptor-2 association of EIF4EBP1 with RPTOR and FRAP1 906882 n/a 906886 unspecified role unspecified role prey prey n-terminal HA tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 906885 unspecified role unspecified role prey prey region flag tagged 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 false n/a 906887 unspecified role unspecified role bait bait region GST tagged 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 association association false false false Fig. 4. C Q8N122;Q9JLN9;B2R502 transfection frap1-rptor-3 physical association of FRAP1 with RPTOR 906883 n/a 906889 unspecified role unspecified role bait bait n/a 906885 unspecified role unspecified role prey prey physical association physical association false false false Fig. 2. B and fig. 6. A specific monoclonal antibody against FRAP1 for the co-immunoprecipitation and specific polyclonal antibodies against RPTOR for the immuno blot rptor-frap1-2 physical association of RPTOR with FRAP1 906883 n/a 906889 unspecified role unspecified role prey prey n/a 906885 unspecified role unspecified role bait bait physical association physical association false false false Fig. 2. B specific polyclonal antibodies antibody against RPTOR for the co-immunoprecipitation and specific monoclonal antibody against FRAP1 for the immuno blot The immunoprecipitate was subjected to the kinase assay using GST-4EBP1 as a substrate.The anti-N-raptor Ab precipitates a kinase activity toward GST-4EBP1. frap1-rptor-1 physical association of FRAP1 with RPTOR 906884 n/a 906885 unspecified role unspecified role prey prey n/a 906889 unspecified role unspecified role bait bait physical association physical association false false false Fig. 1. B