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/ DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. Cell (0092-8674) 2009 imex curation Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu davet Only protein-protein interactions 15 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 Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu Accepted 2009-SEP-5 by DAVET. Cell (0092-8674) 2009 davet Only protein-protein interactions 15 imex curation DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. Cell (0092-8674) 2009 imex curation Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu davet Only protein-protein interactions 15 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 de novo protein sequence de novo protein sequencing by mass spectrometry de novo protein sequence Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu Accepted 2009-SEP-5 by DAVET. Cell (0092-8674) 2009 davet Only protein-protein interactions 15 imex curation DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. Cell (0092-8674) 2009 imex curation Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu davet Only protein-protein interactions 15 human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu Accepted 2009-SEP-5 by DAVET. Cell (0092-8674) 2009 davet Only protein-protein interactions 15 imex curation DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. Cell (0092-8674) 2009 imex curation Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu davet Only protein-protein interactions 15 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 Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. Additional information on source of cDNA constructs supplied by the authors sabatini@wi.mit.edu Accepted 2009-SEP-5 by DAVET. Cell (0092-8674) 2009 davet Only protein-protein interactions 15 imex curation DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival. Cell (0092-8674) 2009 imex curation Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu davet Only protein-protein interactions 15 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 de novo protein sequence de novo protein sequencing by mass spectrometry de novo protein sequence Peterson TR., Laplante M., Thoreen CC., Sancak Y., Kang SA., Kuehl WM., Gray NS., Sabatini DM. Immunoprecipitate performed under low salt conditions. sabatini@wi.mit.edu Accepted 2009-SEP-5 by DAVET. Cell (0092-8674) 2009 davet Only protein-protein interactions 15 imex curation rictr_human Rapamycin-insensitive companion of mTOR RICTOR KIAA1999 AVO3 homolog protein protein human Homo sapiens Human MAAIGRGRSLKNLRVRGRNDSGEENVPLDLTREPSDNLREILQNVARLQGVSNMRKLGHLNNFTKLLCDIGHSEEKLGFHYEDIIICLRLALLNEAKEVRAAGLRALRYLIQDSSILQKVLKLKVDYLIARCIDIQQSNEVERTQALRLVRKMITVNASLFPSSVTNSLIAVGNDGLQERDRMVRACIAIICELALQNPEVVALRGGLNTILKNVIDCQLSRINEALITTILHLLNHPKTRQYVRADVELERILAPYTDFHYRHSPDTAEGQLKEDREARFLASKMGIIATFRSWAGIINLCKPGNSGIQSLIGVLCIPNMEIRRGLLEVLYDIFRLPLPVVTEEFIEALLSVDPGRFQDSWRLSDGFVAAEAKTILPHRARSRPDLMDNYLALILSAFIRNGLLEGLVEVITNSDDHISVRATILLGELLHMANTILPHSHSHHLHCLPTLMNMAASFDIPKEKRLRASAALNCLKRFHEMKKRGPKPYSLHLDHIIQKAIATHQKRDQYLRVQKDIFILKDTEEALLINLRDSQVLQHKENLEWNWNLIGTILKWPNVNLRNYKDEQLHRFVRRLLYFYKPSSKLYANLDLDFAKAKQLTVVGCQFTEFLLESEEDGQGYLEDLVKDIVQWLNASSGMKPERSLQNNGLLTTLSQHYFLFIGTLSCHPHGVKMLEKCSVFQCLLNLCSLKNQDHLLKLTVSSLDYSRDGLARVILSKILTAATDACRLYATKHLRVLLRANVEFFNNWGIELLVTQLHDKNKTISSEALDILDEACEDKANLHALIQMKPALSHLGDKGLLLLLRFLSIPKGFSYLNERGYVAKQLEKWHREYNSKYVDLIEEQLNEALTTYRKPVDGDNYVRRSNQRLQRPHVYLPIHLYGQLVHHKTGCHLLEVQNIITELCRNVRTPDLDKWEEIKKLKASLWALGNIGSSNWGLNLLQEENVIPDILKLAKQCEVLSIRGTCVYVLGLIAKTKQGCDILKCHNWDAVRHSRKHLWPVVPDDVEQLCNELSSIPSTLSLNSESTSSRHNSESESVPSSMFILEDDRFGSSSTSTFFLDINEDTEPTFYDRSGPIKDKNSFPFFASSKLVKNRILNSLTLPNKKHRSSSDPKGGKLSSESKTSNRRIRTLTEPSVDFNHSDDFTPISTVQKTLQLETSFMGNKHIEDTGSTPSIGENDLKFTKNFGTENHRENTSRERLVVESSTSSHMKIRSQSFNTDTTTSGISSMSSSPSRETVGVDATTMDTDCGSMSTVVSTKTIKTSHYLTPQSNHLSLSKSNSVSLVPPGSSHTLPRRAQSLKAPSIATIKSLADCNFSYTSSRDAFGYATLKRLQQQRMHPSLSHSEALASPAKDVLFTDTITMKANSFESRLTPSRFMKALSYASLDKEDLLSPINQNTLQRSSSVRSMVSSATYGGSDDYIGLALPVDINDIFQVKDIPYFQTKNIPPHDDRGARAFAHDAGGLPSGTGGLVKNSFHLLRQQMSLTEIMNSIHSDASLFLESTEDTGLQEHTDDNCLYCVCIEILGFQPSNQLSAICSHSDFQDIPYSDWCEQTIHNPLEVVPSKFSGISGCSDGVSQEGSASSTKSTELLLGVKTIPDDTPMCRILLRKEVLRLVINLSSSVSTKCHETGLLTIKEKYPQTFDDICLYSEVSHLLSHCTFRLPCRRFIQELFQDVQFLQMHEEAEAVLATPPKQPIVDTSAES DIP protein Q6R327 original sequence version: 116 DIP protein Q6R327 original sequence version: 81 DIP protein Q6R327 original sequence version: 93 DB7B1E4A45DAE2AB dptor_human DEP domain-containing mTOR-interacting protein DEPTOR DEP domain-containing protein 6 DEPDC6 protein protein human Homo sapiens Human MEEGGSTGSAGSDSSTSGSGGAQQRELERMAEVLVTGEQLRLRLHEEKVIKDRRHHLKTYPNCFVAKELIDWLIEHKEASDRETAIKLMQKLADRGIIHHVCDEHKEFKDVKLFYRFRKDDGTFPLDNEVKAFMRGQRLYEKLMSPENTLLQPREEEGVKYERTFMASEFLDWLVQEGEATTRKEAEQLCHRLMEHGIIQHVSNKHPFVDSNLLYQFRMNFRRRRRLMELLNEKSPSSQETHDSPFCLRKQSHDNRKSTSFMSVSPSKEIKIVSAVRRSSMSSCGSSGYFSSSPTLSSSPPVLCNPKSVLKRPVTSEELLTPGAPYARKTFTIVGDAVGWGFVVRGSKPCHIQAVDPSGPAAAAGMKVCQFVVSVNGLNVLHVDYRTVSNLILTGPRTIVMEVMEELEC AD132F0E9AFFB90C lst8_human Target of rapamycin complex subunit LST8 MLST8 GBL G protein beta subunit-like LST8 Mammalian lethal with SEC13 protein 8 protein protein human Homo sapiens Human MNTSPGTVGSDPVILATAGYDHTVRFWQAHSGICTRTVQHQDSQVNALEVTPDRSMIAAAGYQHIRMYDLNSNNPNPIISYDGVNKNIASVGFHEDGRWMYTGGEDCTARIWDLRSRNLQCQRIFQVNAPINCVCLHPNQAELIVGDQSGAIHIWDLKTDHNEQLIPEPEVSITSAHIDPDASYMAAVNSTGNCYVWNLTGGIGDEVTQLIPKTKIPAHTRYALQCRFSPDSTLLATCSADQTCKIWRTSNFSLMTELSIKSGNPGESSRGWMWGCAFSGDSQYIVTASSDNLARLWCVETGEIKREYGGHQKAVVCLAFNDSVLG 43A600D4EF2B6543 akts1_human Proline-rich AKT1 substrate 1 40 kDa proline-rich AKT substrate AKT1S1 PRAS40 protein protein human Homo sapiens Human MASGRPEELWEAVVGAAERFRARTGTELVLLTAAPPPPPRPGPCAYAAHGRGALAEAARRCLHDIALAHRAATAARPPAPPPAPQPPSPTPSPPRPTLAREDNEEDEDEPTETETSGEQLGISDNGGLFVMDEDATLQDLPPFCESDPESTDDGSLSEETPAGPPTCSVPPASALPTQQYAKSLPVSVPVWGFKEKRTEARSSDEENGPPSSPDLDRIAASMRALVLREAEDTQVFGDLPRPRLNTSDFQKLKRKY F6CB195CBB54326C 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 DIP protein P42345 original sequence version: 158 DIP protein P42345 original sequence version: 82 DIP protein P42345 original sequence version: 108 DIP protein P42345 original sequence version: 107 DIP protein P42345 original sequence version: 138 DIP protein P42345 original sequence version: 168 DIP protein P42345 original sequence version: 173 DIP protein P42345 original sequence version: 120 DIP protein P42345 original sequence version: 149 DIP protein P42345 original sequence version: 134 DIP protein P42345 original sequence version: 148 DIP protein P42345 original sequence version: 118 7D9AD6E784882AB4 rptor_human Regulatory-associated protein of mTOR p150 target of rapamycin (TOR)-scaffold protein RPTOR KIAA1303 RAPTOR protein protein human Homo sapiens Human MESEMLQSPLLGLGEEDEADLTDWNLPLAFMKKRHCEKIEGSKSLAQSWRMKDRMKTVSVALVLCLNVGVDPPDVVKTTPCARLECWIDPLSMGPQKALETIGANLQKQYENWQPRARYKQSLDPTVDEVKKLCTSLRRNAKEERVLFHYNGHGVPRPTVNGEVWVFNKNYTQYIPLSIYDLQTWMGSPSIFVYDCSNAGLIVKSFKQFALQREQELEVAAINPNHPLAQMPLPPSMKNCIQLAACEATELLPMIPDLPADLFTSCLTTPIKIALRWFCMQKCVSLVPGVTLDLIEKIPGRLNDRRTPLGELNWIFTAITDTIAWNVLPRDLFQKLFRQDLLVASLFRNFLLAERIMRSYNCTPVSSPRLPPTYMHAMWQAWDLAVDICLSQLPTIIEEGTAFRHSPFFAEQLTAFQVWLTMGVENRNPPEQLPIVLQVLLSQVHRLRALDLLGRFLDLGPWAVSLALSVGIFPYVLKLLQSSARELRPLLVFIWAKILAVDSSCQADLVKDNGHKYFLSVLADPYMPAEHRTMTAFILAVIVNSYHTGQEACLQGNLIAICLEQLNDPHPLLRQWVAICLGRIWQNFDSARWCGVRDSAHEKLYSLLSDPIPEVRCAAVFALGTFVGNSAERTDHSTTIDHNVAMMLAQLVSDGSPMVRKELVVALSHLVVQYESNFCTVALQFIEEEKNYALPSPATTEGGSLTPVRDSPCTPRLRSVSSYGNIRAVATARSLNKSLQNLSLTEESGGAVAFSPGNLSTSSSASSTLGSPENEEHILSFETIDKMRRASSYSSLNSLIGVSFNSVYTQIWRVLLHLAADPYPEVSDVAMKVLNSIAYKATVNARPQRVLDTSSLTQSAPASPTNKGVHIHQAGGSPPASSTSSSSLTNDVAKQPVSRDLPSGRPGTTGPAGAQYTPHSHQFPRTRKMFDKGPEQTADDADDAAGHKSFISATVQTGFCDWSARYFAQPVMKIPEEHDLESQIRKEREWRFLRNSRVRRQAQQVIQKGITRLDDQIFLNRNPGVPSVVKFHPFTPCIAVADKDSICFWDWEKGEKLDYFHNGNPRYTRVTAMEYLNGQDCSLLLTATDDGAIRVWKNFADLEKNPEMVTAWQGLSDMLPTTRGAGMVVDWEQETGLLMSSGDVRIVRIWDTDREMKVQDIPTGADSCVTSLSCDSHRSLIVAGLGDGSIRVYDRRMALSECRVMTYREHTAWVVKASLQKRPDGHIVSVSVNGDVRIFDPRMPESVNVLQIVKGLTALDIHPQADLIACGSVNQFTAIYNSSGELINNIKYYDGFMGQRVGAISCLAFHPHWPHLAVGSNDYYISVYSVEKRVR DIP protein Q8N122 original sequence version: 79 DIP protein Q8N122 original sequence version: 129 DIP protein Q8N122 original sequence version: 138 DIP protein Q8N122 original sequence version: 142 DIP protein Q8N122 original sequence version: 91 DIP protein Q8N122 original sequence version: 118 DIP protein Q8N122-1 original sequence version: 89 DIP protein Q8N122 original sequence version: 128 688ED1943F45045A mtor_rat Serine/threonine-protein kinase mTOR FK506-binding protein 12-rapamycin complex-associated protein 1 Rapamycin target protein 1 Mtor Mammalian target of rapamycin Frap1 Raft1 FKBP12-rapamycin complex-associated protein Mechanistic target of rapamycin protein protein rat Rattus norvegicus (Rat) Rat MLGTGPATATAGAATSSNVSVLQQFASGLKSRNEETRAKAAKELQHYVTMELREMSQEESTRFYDQLNHHIFELVSSSDANERKGGILAIASLIGVEGGNSTRIGRFANYLRNLLPSSDPVVMEMASKAIGRLAMAGDTFTAEYVEFEVKRALEWLGADRNEGRRHAAVLVLRELAISVPTFFFQQVQPFFDNIFVAVWDPKQAIREGAVAALRACLILTTQREPKEMQKPQWYRHTFEEAEKGFDETLAKEKGMNRDDRIHGALLILNELVRISSMEGERLREEMEEITQQQLVHDKYCKDLMGFGTKPRHITPFTSFQAVQPQQSNALVGLLGYSSHQGLMGFGASPSPTKSTLVESRCCRDLMEEKFDQVCQWVLKCRSSKNSLIQMTILNLLPRLAAFRPSAFTDTQYLQDTMNHVLSCVKKEKERTAAFQALGLLSVAVRSEFKVYLPRVLDIIRAALPPKDFAHKRQKTVQVDATVFTCISMLARAMGPGIQQDIKELLEPMLAVGLSPALTAVLYDLSRQIPQLKKDIQDGLLKMLSLVLMHKPLRHPGMPKGLAHQLASPGLTTLPEASDVASITLALRTLGSFEFEGHSLTQFVRHCADHFLNSEHKEIRMEAARTCSRLLTPSIHLISGHAHVVSQTAVQVVADVLSKLLVVGITDPDPDIRYCVLASLDERFDAHLAQAENLQALFVALNDQVFEIRELAICTVGRLSSMNPAFVMPFLRKMLIQILTELEHSGIGRIKEQSARMLGHLVSNAPRLIRPYMEPILKALILKLKDPDPDPNPGVINNVLATIGELAQVSGLEMRKWVDELFVIIMDMLQDSSLLAKRQVALWTLGQLVASTGYVVEPYRKYPTLLEVLLNFLKTEQNQGTRREAIRVLGLLGALDPYKHKVNIGMIDQSRDASAVSLSESKSSQDSSDYSTSEMLVNMGNLPLDEFYPAVSMVALMRIFRDQSLSHHHTMVVQAITFIFKSLGLKCVQFLPQVMPTFLNVIRVCDGAIREFLFQQLGMLVSFVKSHIRPYMDEIVTLMREFWVMNTSIQSTIILLIEQIVVALGGEFKLYLPQLIPHMLRVFMHDNSQGRIVSIKLLAAIQLFGANLDDYLHLLLPPIVKLFDAPEVPLPSRKAALETVDRLTESLDFTDYASRIIHPIVRTLDQSPELRSTAMDTLSSLVFQLGKKYQIFIPMVNKVLVRHRINHQRYDVLICRIVKGYTLADEEEDPLIYQHRMLRSSQGDALASGPVETGPMKKLHVSTINLQKAWGAARRVSKDDWLEWLRRLSLELLKDSSSPSLRSCWALAQAYNPMARDLFNAAFVSCWSELNEDQQDELIRSIELALTSQDIAEVTQTLLNLAEFMEHSDKGPLPLRDDNGIVLLGERAAKCRAYAKALHYKELEFQKGPTPAILESLISINNKLQQPEAASGVLEYAMKHFGELEIQATWYEKLHEWEDALVAYDKKMDTNKDDPELMLGRMRCLEALGEWGQLHQQCCEKWTLVNDETQAKMARMAAAAAWGLGQWDSMEEYTCMIPRDTHDGAFYRAVLALHQDLFSLAQQCIDKARDLLDAELTAMAGESYSRAYGAMVSCHMLSELEEVIQYKLVPERREIIRQIWWERLQGCQRIVEDWQKILMVRSLVVSPHEDMRTWLKYASLCGKSGRLALAHKTLVLLLGVDPSRQLDHPLPTVHPQVTYAYMKNMWKSARKIDAFQHMQHFVQTMQQQAQHAIATEDQQHKQELHKLMARCFLKLGEWQLNLQGINESTIPKVLQYYSAATEHDRSWYKAWHAWAVMNFEAVLHYKHQNQARDEKKKLRHASGANITNATTTATTAASAAAATSTEGSNSESEAESNESSPTPSPLQKKVTEDLSKTLLLYTVPAVQGFFRSISLSRGNNLQDTLRVLTLWFDYGHWPDVNEALVEGVKAIQIDTWLQVIPQLIARIDTPRPLVGRLIHQLLTDIGRYHPQALIYPLTVASKSTTTARHNAANKILKNMCEHSNTLVQQAMMVSEELIRVAILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQLPQLTSLELQYVSPKLLMCRDLELAVPGTYDPNQPIIRIQSIAPSLQVITSKQRPRKLTLMGSNGHEFVFLLKGHEDLRQDERVMQLFGLVNTLLANDPTSLRKNLSIQRYAVIPLSTNSGLIGWVPHCDTLHALIRDYREKKKILLNIEHRIMLRMAPDYDHLTLMQKVEVFEHAVNNTAGDDLAKLLWLKSPSSEVWFDRRTNYTRSLAVMSMVGYILGLGDRHPSNLMLDRLSGKILHIDFGDCFEVAMTREKFPEKIPFRLTRMLTNAMEVTGLDGNYRTTCHTVMEVLREHKDSVMAVLEAFVYDPLLNWRLMDTNAKGNKRSRTRTDSYSAGQSVEILDGVELGEPAHKKTGTTVPESIHSFIGDGLVKPEALNKKAIQIINRVRDKLTGRDFSHDDTLDVPTQVELLIKQATSHENLCQCYIGWCPFW DIP protein P42346 original sequence version: 147 BE841EA7B9086F99 frap1-depdc6-1 5 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Using low-salt purification conditions designed to isolate PRAS40, we identified within mTOR immunoprecipitates a 48 kDa protein assigned the NCBI gene symbol DEPDC6 (NCBI gene ID 64798) (Figure 1A). Fig. 1A frap1-depdc6-2 2 6 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 7 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 10 unspecified role unspecified role prey prey physical association physical association In purified preparations of recombinant DEPTOR stably expressed in HEK293E cells, we detected via mass spectrometry endogenous mTOR, as well as raptor and rictor, mTORC1, and mTORC2-specific components, respectively. Fig. 1C frap1-akt1s1-1 2 11 unspecified role unspecified role prey prey 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 10 unspecified role unspecified role prey prey physical association physical association Analogous preparations of recombinant PRAS40, a raptor binding protein, contained only mTORC1 (Figure 1C) Fig. 1C frap1-depdc6-3 1 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Consistent with DEPTOR being part of mTORC1 and mTORC2, endogenous DEPTOR was also detected in immunoprecipitates prepared from HEK293E (Figure 1D) or HeLa cells (Figure 1E) with antibodies that recognize endogenous mTOR, raptor, or rictor, but not actin. Fig. 1D Increased salt concentrations reduced the amounts of DEPTOR recovered in the immunoprecipitates. frap1-depdc6-4 1 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association Consistent with DEPTOR being part of mTORC1 and mTORC2, endogenous DEPTOR was also detected in immunoprecipitates prepared from HEK293E (Figure 1D) or HeLa cells (Figure 1E) with antibodies that recognize endogenous mTOR, raptor, or rictor, but not actin. Fig. 1D Increased salt concentrations reduced the amounts of DEPTOR recovered in the immunoprecipitates. frap1-depdc6-5 1 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey physical association physical association Consistent with DEPTOR being part of mTORC1 and mTORC2, endogenous DEPTOR was also detected in immunoprecipitates prepared from HEK293E (Figure 1D) or HeLa cells (Figure 1E) with antibodies that recognize endogenous mTOR, raptor, or rictor, but not actin. Fig. 1D Increased salt concentrations reduced the amounts of DEPTOR recovered in the immunoprecipitates. frap1-depdc6-6 3 6 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey physical association physical association Consistent with DEPTOR being part of mTORC1 and mTORC2, endogenous DEPTOR was also detected in immunoprecipitates prepared from HEK293E (Figure 1D) or HeLa cells (Figure 1E) with antibodies that recognize endogenous mTOR, raptor, or rictor, but not actin. Fig. 1E frap1-depdc6-7 3 11 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey physical association physical association Consistent with DEPTOR being part of mTORC1 and mTORC2, endogenous DEPTOR was also detected in immunoprecipitates prepared from HEK293E (Figure 1D) or HeLa cells (Figure 1E) with antibodies that recognize endogenous mTOR, raptor, or rictor, but not actin. Fig. 1E frap1-depdc6-8 3 10 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association Consistent with DEPTOR being part of mTORC1 and mTORC2, endogenous DEPTOR was also detected in immunoprecipitates prepared from HEK293E (Figure 1D) or HeLa cells (Figure 1E) with antibodies that recognize endogenous mTOR, raptor, or rictor, but not actin. Fig. 1E frap1-depdc6-9 3 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Consistent with DEPTOR being part of mTORC1 and mTORC2, endogenous DEPTOR was also detected in immunoprecipitates prepared from HEK293E (Figure 1D) or HeLa cells (Figure 1E) with antibodies that recognize endogenous mTOR, raptor, or rictor, but not actin. Fig. 1E frap-depdc6-1 4 8 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 association association To test this, we co-overexpressed DEPTOR with epitope-tagged mTOR or mLST8/GβL and analyzed mTOR or mLST8/GβL immunoprecipitates for the presence of DEPTOR. Fig. 1F frap-depdc6-2 4 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 12 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 8 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 association association To test this, we co-overexpressed DEPTOR with epitope-tagged mTOR or mLST8/GβL and analyzed mTOR or mLST8/GβL immunoprecipitates for the presence of DEPTOR. Fig. 1F frap-depdc6-3 4 12 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 physical association physical association DEPTOR interacts with a C-terminal portion of mTOR (aa 1483�2000) Fig. 1F, 1G frap1-depdc6-10 1 6 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey 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 undefined mutation mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:48:08 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'ser_thr-ala' association association Compared to wild-type DEPTOR, the 13�S/T→A DEPTOR mutant bound better to mTOR within mTORC1 and mTORC2 (Figure 5A). Fig. 5A frap1-depdc6-11 1 11 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey 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 undefined mutation mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:47:40 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'ser_thr-ala' association association Compared to wild-type DEPTOR, the 13�S/T→A DEPTOR mutant bound better to mTOR within mTORC1 and mTORC2 (Figure 5A). Fig. 5A