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/ NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. NSAID Sulindac and Its Analog Bind RXRalpha and Inhibit RXRalpha-Dependent AKT Signaling. Cancer Cell (1535-6108) 2010 imex curation 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 13-Sept-2010: Contacted by JYOTI. orchard Only protein-protein interactions 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 For this Expt 'Pull down' came from the Kollari paper also. [The species of RXRa is not stated in this paper, but an earlier paper] (Kolluri et al.,2005) refers to human RXRa LBD (223�462) prepared as a polyhistidine-tagged fusion protein. (the authors responded very quickly if you think there is need to further clarify this.) 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 Accepted 2010-AUG-6 by ORCHARD. 13-Sept-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation 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 sulindac Sulindac, an NSAID small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 3-h_alitretinoin Tritiated Alitretinoin, (9-cis)-retinoic acid. small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 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. 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 DIP protein P27986 original sequence version: 181 DIP protein P27986 original sequence version: 184 DIP protein P27986-1 original sequence version: 135 DIP protein P27986 original sequence version: 135 DIP protein P27986-1 original sequence version: 145 DIP protein P27986-1 original sequence version: 147 DIP protein P27986 original sequence version: 191 B9DAD8416C33140F 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 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 DIP protein P19793 original sequence version: 177 DIP protein P19793 original sequence version: 107 DIP protein P19793 original sequence version: 153 DIP protein P19793 original sequence version: 131 DIP protein P19793 original sequence version: 136 DIP protein P19793 original sequence version: 169 DIP protein P19793 original sequence version: 161 DIP protein P19793 original sequence version: 139 7F952B580AD84C42 rxra-sulindac 1 13 unspecified role unspecified role prey prey 14 competitor competitor competitor prey prey 18 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 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 physical association physical association Sulindac bound to RXRa, but not RAR, with an IC50 of 80 uM. Fig 1A. (9-cis)-retinoic acid was radiolabelled. rxra-sulindac-1 8 13 unspecified role unspecified role neutral component neutral component 18 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 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 direct interaction direct interaction 19F nuclear magnetic resonance (NMR) spectra showed that the signal intensity of the fluorine spectrum of Sulindac was strongly suppressed by the RXRa LBD but not by Nur77 protein, demonstrating a direct and specific binding. Fig 1D. rxra-p85a-1 2 18 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey physical association physical association 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-p85a 4 18 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 rxra-d80 RXRa- D80 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 16 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 physical association physical association 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-sulindac-2 3 13 unspecified role unspecified role prey prey 18 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 physical association physical association Sulindac binding to RXRa in cells. Figs 1B, S1B. rxra-p85a-2 4 16 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 18 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 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 physical association physical association RXRa/1-134 but not RXRa/223-462 could interact with p85a. Fig 5B. "modulating-domain" taken from SwissP - no relevant domain name in Interpro for the exact aa nos. I have entered the X Ref IPR021780 - which is aas 17 - 127, a "Protein of unknown function DUF3345". Should this X ref be removed? rxra-p85a-3 6 18 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 region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 16 unspecified role unspecified role neutral component neutral component colocalization colocalization 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 7 18 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 region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 16 unspecified role unspecified role neutral component neutral component colocalization colocalization 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-k80003 8 15 unspecified role unspecified role neutral component neutral component 18 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 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 direct interaction direct interaction Fig 7E. The binding of K-80003 to RXRα was also confirmed by 19F NMR binding assays (Figure 7E) p85a-rxra 5 18 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 region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 16 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 physical association physical association 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 9 16 unspecified role unspecified role prey prey 18 unspecified role unspecified role bait bait physical association physical association 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 10 17 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey physical association physical association 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. There seem to be no UniProt or ENA entries for Hamster RXRA proteins, and no UNiProt entry for p85a-hamster - did not bother to look in ENA for this: so I have created entries in the protein editor ... p85a-rxra-3 11 18 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey physical association physical association 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).