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/ Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Cancer Cell (1535-6108) 2013 imex curation Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov 01-March-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay western blot western blot Immuno blot Immuno blot Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov Accepted 2013-FEB-20 by ORCHARD 01-March-2013: Contacted by IntAct-Help. Cancer Cell (1535-6108) 2013 Only protein-protein interactions imex curation Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Cancer Cell (1535-6108) 2013 imex curation Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov 01-March-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov Accepted 2013-FEB-20 by ORCHARD 01-March-2013: Contacted by IntAct-Help. Cancer Cell (1535-6108) 2013 Only protein-protein interactions imex curation Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Cancer Cell (1535-6108) 2013 imex curation Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov 01-March-2013: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov Accepted 2013-FEB-20 by ORCHARD 01-March-2013: Contacted by IntAct-Help. Cancer Cell (1535-6108) 2013 Only protein-protein interactions imex curation Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. Cancer Cell (1535-6108) 2013 imex curation Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov 01-March-2013: Contacted by IntAct-Help. Only protein-protein interactions human-786_o Homo sapiens renal cell adenocarcinoma 786_o Human renal cell adenocarcinoma cell line CRL-1932 is the ATCC identifier. ATCC identifier is CRL-1932. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sourbier C., Scroggins B.T., Ratnayake R., Prince T.L., Lee S., Lee M.J., Nagy P.L., Lee Y.H., Trepel J.B., Beutler J.A., Linehan W.M., Neckers L. Cancer - Interactions investigated in the context of cancer The shift of fluorescence emission to 420 nm was measured to determine SAPD binding/reduction of EA binding to PKC proteins. Interactions involving the use of "PKC activity assay from Enzo Life Sciences" (Kinase Assay) cannot be entered as the reaction substrate is only described as a "peptide" and Enzo does not wish to further identify it. Interactions involving HSP90 have not been included due to lack of antibody specificity. The authors have confirmed that the IRS1 Kinase assay substrate is Rattine in origin. (Millipore 13-124). neckersl@mail.nih.gov Accepted 2013-FEB-20 by ORCHARD 01-March-2013: Contacted by IntAct-Help. Cancer Cell (1535-6108) 2013 Only protein-protein interactions imex curation irs1_rat Insulin receptor substrate 1 pp185 Irs1 Irs-1 protein protein rat Rattus norvegicus (Rat) Rat MASPPDTDGFSDVRKVGYLRKPKSMHKRFFVLRAASEAGGPARLEYYENEKKWRHKSSAPKRSIPLESCFNINKRADSKNKHLVALYTRDEHFAIAADSEAEQDSWYQALLQLHNRAKAHHDGAGGGCGGSCSGSSGVGEAGEDLSYDTGPGPAFKEVWQVILKPKGLGQTKNLIGIYRLCLTSKTISFVKLNSEAAAVVLQLMNIRRCGHSENFFFIEVGRSAVTGPGEFWMQVDDSVVAQNMHETILEAMRAMSDEFRPRTKSQSSSSCSNPISVPLRRHHLNNPPPSQVGLTRRSRTESITATSPASMVGGKPGSFRVRASSDGEGTMSRPASVDGSPVSPSTNRTHAHRHRGSSRLHPPLNHSRSIPMPSSRCSPSATSPVSLSSSSTSGHGSTSDCLFPRRSSASVSGSPSDGGFISSDEYGSSPCDFRSSFRSVTPDSLGHTPPARGEEELSNYICMGGKGASTLTAPNGHYILSRGGNGHRYIPGATMGTSPALTGDEAAGAADLDNRFRKRTHSAGTSPTISHQKTPSQSSVVSIEEYTEMMPAAYPPGGGSGGRLPGYRHSAFVPTHSYPEEGLEMHHLERRGGHHRPDSSNLHTDDGYMPMSPGVAPVPSNRKGNGDYMPMSPKSVSAPQQIINPIRRHPQRVDPNGYMMMSPSGSCSPDIGGGSCSSSSISAAPSGSSYGKPWTNGVGGHHTHALPHAKPPVESGGGKLLPCTGDYMNMSPVGDSNTSSPSECYYGPEDPQHKPVLSYYSLPRSFKHTQRPGEPEEGARHQHLRLSSSSGRLRYTATAEDSSSSTSSDSLGGGYCGARPESSVTHPHHHALQPHLPRKVDTAAQTNSRLARPTRLSLGDPKASTLPRVREQQQQQQQQQQSSLHPPEPKSPGEYVNIEFGSGQPGYLAGPATSRSSPSVRCLPQLHPAPREETGSEEYMNMDLGPGRRATWQESGGVELGRVGPAPPGAASICRPTRSVPNSRGDYMTMQIGCPRQSYVDTSPVAPVSYADMRTGIAAEKVSLPRTTGAAPPPSSTASASASVTPQGAAEQAAHSSLLGGPQGPGGMSAFTRVNLSPNHNQSAKVIRADTQGCRRRHSSETFSAPTRAANTVSFGAGAAGGGSGGGSEDVKRHSSASFENVWLRPGDLGGASKESAPGCGAAGGLEKSLNYIDLDLVKDVKQHPQDCPSQQQSLPPPPPHQPLGSNEGSSPRRSSEDLSTYASINFQKQPEDRQ A274BC7540CA85C5 kpct_human Protein kinase C theta type PRKCQ nPKC-theta PRKCT protein protein human Homo sapiens Human MSPFLRIGLSNFDCGSCQSCQGEAVNPYCAVLVKEYVESENGQMYIQKKPTMYPPWDSTFDAHINKGRVMQIIVKGKNVDLISETTVELYSLAERCRKNNGKTEIWLELKPQGRMLMNARYFLEMSDTKDMNEFETEGFFALHQRRGAIKQAKVHHVKCHEFTATFFPQPTFCSVCHEFVWGLNKQGYQCRQCNAAIHKKCIDKVIAKCTGSAINSRETMFHKERFKIDMPHRFKVYNYKSPTFCEHCGTLLWGLARQGLKCDACGMNVHHRCQTKVANLCGINQKLMAEALAMIESTQQARCLRDTEQIFREGPVEIGLPCSIKNEARPPCLPTPGKREPQGISWESPLDEVDKMCHLPEPELNKERPSLQIKLKIEDFILHKMLGKGSFGKVFLAEFKKTNQFFAIKALKKDVVLMDDDVECTMVEKRVLSLAWEHPFLTHMFCTFQTKENLFFVMEYLNGGDLMYHIQSCHKFDLSRATFYAAEIILGLQFLHSKGIVYRDLKLDNILLDKDGHIKIADFGMCKENMLGDAKTNTFCGTPDYIAPEILLGQKYNHSVDWWSFGVLLYEMLIGQSPFHGQDEEELFHSIRMDNPFYPRWLEKEAKDLLVKLFVREPEKRLGVRGDIRQHPLFREINWEELERKEIDPPFRPKVKSPFDCSNFDKEFLNEKPRLSFADRALINSMDQNMFRNFSFMNPGMERLIS B3C53AB892D5210A sapintoxin_d SAPINTOXIN_D small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) hsf1_human Heat shock factor protein 1 Heat shock transcription factor 1 HSF1 HSTF1 protein protein human Homo sapiens Human MDLPVGPGAAGPSNVPAFLTKLWTLVSDPDTDALICWSPSGNSFHVFDQGQFAKEVLPKYFKHNNMASFVRQLNMYGFRKVVHIEQGGLVKPERDDTEFQHPCFLRGQEQLLENIKRKVTSVSTLKSEDIKIRQDSVTKLLTDVQLMKGKQECMDSKLLAMKHENEALWREVASLRQKHAQQQKVVNKLIQFLISLVQSNRILGVKRKIPLMLNDSGSAHSMPKYSRQFSLEHVHGSGPYSAPSPAYSSSSLYAPDAVASSGPIISDITELAPASPMASPGGSIDERPLSSSPLVRVKEEPPSPPQSPRVEEASPGRPSSVDTLLSPTALIDSILRESEPAPASVTALTDARGHTDTEGRPPSPPPTSTPEKCLSVACLDKNELSDHLDAMDSNLDNLQTMLSSHGFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPRPPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAEDPTISLLTGSEPPKAKDPTVS DIP protein Q00613 original sequence version: 97 DIP protein Q00613 original sequence version: 157 DIP protein Q00613 original sequence version: 161 DIP protein Q00613-1 original sequence version: 161 735074507C954365 englerin_a ENGLERIN A small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) irs1-prkcq 2 5 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 6 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range phosphorylation phosphorylation reaction phosphorylation 1C PKC-theta was produced in Insect cells. EA - Englerin A irs1-prkcq-1 1 5 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 6 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range phosphorylation phosphorylation reaction phosphorylation 1D PKC-theta was produced in Insect cells. EA - Englerin A prkcq-englerin 3 9 unspecified role unspecified role prey prey 7 competitor competitor competitor prey prey 6 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction 1F PKC-theta was produced in Insect cells. prkcq-hsf1 4 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association 4C HSP90 interaction with the PKCθ/HSF1 immunocomplex was reduced by EA treatment (left panel). HSP90 cannot be entered as an interactor due to lack of antibody specificity. irs1-prkcq-2 2 6 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 8 enzyme target enzyme target substrate substrate neutral component neutral component phosphorylation phosphorylation reaction phosphorylation 4D PKC-theta was produced in Insect cells. EA - Englerin A prkcq-sapd 3 6 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 7 unspecified role unspecified role prey prey direct interaction direct interaction 1F-2 PKC-theta was produced in Insect cells.