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/ zhou-2010-2 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-4 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-1 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-3 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-5 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. tap tandem affinity purification TAP TAP tap predetermined predetermined participant predetermined Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-6 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma confocal microscopy confocal microscopy immunostaining immunostaining Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-7 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-zr-75-1 Human zr-75-1 breast carcinoma cells zr-75-1 ZR-75-1, Human breast ductal carcinoma cell line. Reported to express the oestrogen and progesterone receptors. confocal microscopy confocal microscopy immunostaining immunostaining Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-9 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-a_549 Homo sapiens Caucasian lung carcinoma cell line a_549 Human Caucasian lung carcinoma anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-10 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-nci_h292 Homo sapiens H292 lung carcinoma tissue nci_h292 H292 lung carcinoma tissue mucoepidermoid; Derived from: lymph node metastasis. Often referred in the literature as H-292. mucoepidermoid; Derived from: lymph node metastasis. Often referred in the literature as H-292. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-11 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. mesau-bhk Mesocricetus auratus baby kidney cell line BHK bhk Golden (Syrian) Hamster baby kidney cell line BHK Established from the kidneys of five unsexed, one-day-old hamsters in 1961. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation zhou-2010-12 NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhou H., Liu W., Su Y., Wei Z., Liu J., Kolluri SK., Wu H., Cao Y., Chen J., Wu Y., Yan T., Cao X., Gao W., Molotkov A., Jiang F., Li WG., Lin B., Zhang HP., Yu J., Luo SP., Zeng JZ., Duester G., Huang PQ., Zhang XK. Cancer - Interactions investigated in the context of cancer Proteins purified with InterPlay Mammalian TAP System (Stratagene) were analyzed by HPLC. Sulindac was detected with a photoarray detector. Details of constructs supplied by the authors. xzhang@burnham.org Cancer Cell (1535-6108) 2010 Only protein-protein interactions imex curation sulindac Sulindac, an NSAID small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) rxra_human Retinoic acid receptor RXR-alpha RXRA NR2B1 Retinoid X receptor alpha Nuclear receptor subfamily 2 group B member 1 protein protein human Homo sapiens Human MDTKHFLPLDFSTQVNSSLTSPTGRGSMAAPSLHPSLGPGIGSPGQLHSPISTLSSPINGMGPPFSVISSPMGPHSMSVPTTPTLGFSTGSPQLSSPMNPVSSSEDIKPPLGLNGVLKVPAHPSGNMASFTKHICAICGDRSSGKHYGVYSCEGCKGFFKRTVRKDLTYTCRDNKDCLIDKRQRNRCQYCRYQKCLAMGMKREAVQEERQRGKDRNENEVESTSSANEDMPVERILEAELAVEPKTETYVEANMGLNPSSPNDPVTNICQAADKQLFTLVEWAKRIPHFSELPLDDQVILLRAGWNELLIASFSHRSIAVKDGILLATGLHVHRNSAHSAGVGAIFDRVLTELVSKMRDMQMDKTELGCLRAIVLFNPDSKGLSNPAEVEALREKVYASLEAYCKHKYPEQPGRFAKLLLRLPALRSIGLKCLEHLFFFKLIGDTPIDTFLMEMLEAPHQMT k-80003 K-80003, a Sulindac (NSAID) analog. small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) K-80003 has increased affinity to RXRa (than Sulindac) but lacks COX inhibitory activity. It is also a more potent inhibitor of AKT activation and cancer cell growth. p85a_human Phosphatidylinositol 3-kinase regulatory subunit alpha PIK3R1 GRB1 Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha protein protein human Homo sapiens Human MSAEGYQYRALYDYKKEREEDIDLHLGDILTVNKGSLVALGFSDGQEARPEEIGWLNGYNETTGERGDFPGTYVEYIGRKKISPPTPKPRPPRPLPVAPGSSKTEADVEQQALTLPDLAEQFAPPDIAPPLLIKLVEAIEKKGLECSTLYRTQSSSNLAELRQLLDCDTPSVDLEMIDVHVLADAFKRYLLDLPNPVIPAAVYSEMISLAPEVQSSEEYIQLLKKLIRSPSIPHQYWLTLQYLLKHFFKLSQTSSKNLLNARVLSEIFSPMLFRFSAASSDNTENLIKVIEILISTEWNERQPAPALPPKPPKPTTVANNGMNNNMSLQDAEWYWGDISREEVNEKLRDTADGTFLVRDASTKMHGDYTLTLRKGGNNKLIKIFHRDGKYGFSDPLTFSSVVELINHYRNESLAQYNPKLDVKLLYPVSKYQQDQVVKEDNIEAVGKKLHEYNTQFQEKSREYDRLYEEYTRTSQEIQMKRTAIEAFNETIKIFEEQCQTQERYSKEYIEKFKREGNEKEIQRIMHNYDKLKSRISEIIDSRRRLEEDLKKQAAEYREIDKRMNSIKPDLIQLRKTRDQYLMWLTQKGVRQKKLNEWLGNENTEDQYSLVEDDEDLPHHDEKTWNVGSSNRNKAENLLRGKRDGTFLVRESSKQGCYACSVVVDGEVKHCVINKTATGYGFAEPYNLYSSLKELVLHYQHTSLVQHNDSLNVTLAYPVYAQQRR 3-h_alitretinoin Tritiated Alitretinoin, (9-cis)-retinoic acid. small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) rxra_mesau_protein Retinoic acid receptor RXR-alpha protein protein mesau Mesocricetus auratus (Golden hamster) Golden hamster An amino acid sequence which is not represented in UniProt p85a_mesau_protein Phosphatidylinositol 3-kinase regulatory subunit alpha protein protein mesau Mesocricetus auratus (Golden hamster) Golden hamster An amino acid sequence which is not represented in UniProt rxra-sulindac-1 958305 n/a 958316 unspecified role unspecified role neutral component neutral component n/a 958317 unspecified role unspecified role neutral component neutral component lbd ligand binding 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 direct interaction direct interaction false false false Fig 1D. rxra-k80003 958305 n/a 958318 unspecified role unspecified role neutral component neutral component n/a 958317 unspecified role unspecified role neutral component neutral component lbd ligand binding 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 direct interaction direct interaction false false false Fig 7E. rxra-p85a 958306 n/a 958317 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 false rxra-d80 RXRa- D80 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 958319 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 false physical association physical association false false false Similarly to tRXRa, RXRa/Delta80 interacted with p85a, which was strongly enhanced by TNF-a. In contrast, the full-length RXRa did not interact with p85a either in the absence or presence of TNF-a. Fig 5A. TNF-alpha rxra-p85a-2 958306 n/a 958319 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 false n/a 958317 unspecified role unspecified role prey prey modulating-domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false RXRa/1-134 but not RXRa/223-462 could interact with p85a. Fig 5B. rxra-sulindac 958307 n/a 958316 unspecified role unspecified role prey prey n/a 958320 competitor competitor competitor prey prey n/a 958317 unspecified role unspecified role bait bait lbd ligand binding 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 physical association physical association false false false Sulindac bound to RXRa, but not RAR, with an IC50 of 80 uM. Fig 1A. (9-cis)-retinoic acid was radiolabelled. rxra-p85a-1 958308 n/a 958317 unspecified role unspecified role bait bait n/a 958319 unspecified role unspecified role prey prey physical association physical association false false false The interaction of p85a with RXRa/tRXRa in the presence of TNF-a (or 9-cis-RA) was inhibited by Sulindac. Fig 3E. Sulindac. TNFa. RXRa was precipitated using an antibody that recognizes the C-terminal E/F domain, (amino acids 225-462). rxra-sulindac-2 958309 n/a 958316 unspecified role unspecified role prey prey n/a 958317 unspecified role unspecified role bait bait c terminus tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Sulindac binding to RXRa in cells. Figs 1B, S1B. rxra-p85a-3 958310 n/a 958317 unspecified role unspecified role neutral component neutral component rxra-d80 RXRa- D80 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 958319 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false RXRa/D80 predominantly resided in the cytoplasm, with occasional punctate plasma membrane localization. Thus, deletion of the N-terminal sequences of RXRa alters its subcellular localization and confers its ability to interact with p85a. Fig 5D. rxra-p85a-4 958311 n/a 958317 unspecified role unspecified role neutral component neutral component rxra-d80 RXRa- D80 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 958319 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false RXRa/D80 predominantly resided in the cytoplasm, with occasional punctate plasma membrane localization. Thus, deletion of the N-terminal sequences of RXRa alters its subcellular localization and confers its ability to interact with p85a. Fig 5D. p85a-rxra 958312 n/a 958317 unspecified role unspecified role prey prey rxra-d80 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 958319 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 false physical association physical association false false false The interaction of RXRa/D80 with p85a either in the absence or presence of TNFa was more potently inhibited by K-80003 than by Sulindac. Fig 8C. Sulindac, K-80003. TNFa. p85a-rxra-1 958313 n/a 958319 unspecified role unspecified role prey prey n/a 958317 unspecified role unspecified role bait bait physical association physical association false false false The interaction of p85a with RXRa/tRXRa in the presence of 9-cis-RA or (TNF-a) was inhibited by Sulindac. Fig S3D. Sulindac. 9-cis-RA. RXRa was precipitated using an antibody that recognizes the C-terminal E/F domain, (amino acids 225-462). p85a-rxra-2 958314 n/a 958321 unspecified role unspecified role bait bait n/a 958322 unspecified role unspecified role prey prey physical association physical association false false false Induction of RXRa-p85a interaction by RA. Fig S3E. 9-cis-Retinoic Acid. RXRa was precipitated using an antibody that recognizes the C-terminal E/F domain. p85a-rxra-3 958315 n/a 958317 unspecified role unspecified role bait bait n/a 958319 unspecified role unspecified role prey prey physical association physical association false false false Induction of RXRa-p85a interaction by RA. Fig S3E. 9-cis-Retinoic Acid. RXRa was precipitated using an antibody that recognizes the C-terminal E/F domain, (amino acids 225-462).