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/ sourbier-2013-1 Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. 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 Cancer Cell (1535-6108) 2013 Only protein-protein interactions imex curation sourbier-2013-3 Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. 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 Cancer Cell (1535-6108) 2013 Only protein-protein interactions imex curation sourbier-2013-2 Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. 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 Cancer Cell (1535-6108) 2013 Only protein-protein interactions imex curation sourbier-2013-4 Englerin A Stimulates PKCθ to Inhibit Insulin Signaling and to Simultaneously Activate HSF1: Pharmacologically Induced Synthetic Lethality. 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 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 kpct_human Protein kinase C theta type PRKCQ nPKC-theta PRKCT protein protein human Homo sapiens Human MSPFLRIGLSNFDCGSCQSCQGEAVNPYCAVLVKEYVESENGQMYIQKKPTMYPPWDSTFDAHINKGRVMQIIVKGKNVDLISETTVELYSLAERCRKNNGKTEIWLELKPQGRMLMNARYFLEMSDTKDMNEFETEGFFALHQRRGAIKQAKVHHVKCHEFTATFFPQPTFCSVCHEFVWGLNKQGYQCRQCNAAIHKKCIDKVIAKCTGSAINSRETMFHKERFKIDMPHRFKVYNYKSPTFCEHCGTLLWGLARQGLKCDACGMNVHHRCQTKVANLCGINQKLMAEALAMIESTQQARCLRDTEQIFREGPVEIGLPCSIKNEARPPCLPTPGKREPQGISWESPLDEVDKMCHLPEPELNKERPSLQIKLKIEDFILHKMLGKGSFGKVFLAEFKKTNQFFAIKALKKDVVLMDDDVECTMVEKRVLSLAWEHPFLTHMFCTFQTKENLFFVMEYLNGGDLMYHIQSCHKFDLSRATFYAAEIILGLQFLHSKGIVYRDLKLDNILLDKDGHIKIADFGMCKENMLGDAKTNTFCGTPDYIAPEILLGQKYNHSVDWWSFGVLLYEMLIGQSPFHGQDEEELFHSIRMDNPFYPRWLEKEAKDLLVKLFVREPEKRLGVRGDIRQHPLFREINWEELERKEIDPPFRPKVKSPFDCSNFDKEFLNEKPRLSFADRALINSMDQNMFRNFSFMNPGMERLIS hsf1_human Heat shock factor protein 1 Heat shock transcription factor 1 HSF1 HSTF1 protein protein human Homo sapiens Human MDLPVGPGAAGPSNVPAFLTKLWTLVSDPDTDALICWSPSGNSFHVFDQGQFAKEVLPKYFKHNNMASFVRQLNMYGFRKVVHIEQGGLVKPERDDTEFQHPCFLRGQEQLLENIKRKVTSVSTLKSEDIKIRQDSVTKLLTDVQLMKGKQECMDSKLLAMKHENEALWREVASLRQKHAQQQKVVNKLIQFLISLVQSNRILGVKRKIPLMLNDSGSAHSMPKYSRQFSLEHVHGSGPYSAPSPAYSSSSLYAPDAVASSGPIISDITELAPASPMASPGGSIDERPLSSSPLVRVKEEPPSPPQSPRVEEASPGRPSSVDTLLSPTALIDSILRESEPAPASVTALTDARGHTDTEGRPPSPPPTSTPEKCLSVACLDKNELSDHLDAMDSNLDNLQTMLSSHGFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPRPPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAEDPTISLLTGSEPPKAKDPTVS englerin_a ENGLERIN A small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) sapintoxin_d SAPINTOXIN_D small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) irs1-prkcq 1364589 n/a 1364593 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 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 1364594 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 false phosphorylation phosphorylation reaction phosphorylation false false false 1C PKC-theta was produced in Insect cells. EA - Englerin A irs1-prkcq-2 1364589 n/a 1364594 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 false n/a 1364595 enzyme target enzyme target substrate substrate neutral component neutral component phosphorylation phosphorylation reaction phosphorylation false false false 4D PKC-theta was produced in Insect cells. EA - Englerin A prkcq-englerin 1364590 n/a 1364596 unspecified role unspecified role prey prey n/a 1364597 competitor competitor competitor prey prey n/a 1364594 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 false direct interaction direct interaction false false false 1F PKC-theta was produced in Insect cells. prkcq-sapd 1364590 n/a 1364594 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 false n/a 1364597 unspecified role unspecified role prey prey direct interaction direct interaction false false false 1F-2 PKC-theta was produced in Insect cells. irs1-prkcq-1 1364591 n/a 1364593 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 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 1364594 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 false phosphorylation phosphorylation reaction phosphorylation false false false 1D PKC-theta was produced in Insect cells. EA - Englerin A prkcq-hsf1 1364592 n/a 1364595 unspecified role unspecified role prey prey n/a 1364594 unspecified role unspecified role bait bait physical association physical association false false false 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.