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/ navarro-2012-2 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. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag visualisation tag visualisation Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ., Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ. pmccormick@ub.edu PloS one 2012 Only protein-protein interactions imex curation imex curation navarro-2012-4 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. fluorescence fluorescence technology fluorescence tag visualisation tag visualisation Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ., Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ. SRET experiment:- Sequential BRET-FRET; the oxidation of an Rluc substrate triggers acceptor excitation by BRET and sub- sequent fluorescent energy transfer to a FRET acceptor. This technique permits heteromers formed by three different proteins to be identified. In this instance there is an initial bioluminescent energy transfer between Rluc and a blue shifted GFP2, then a second fluorescent energy transfer event between excited GFP2 and YFP. pmccormick@ub.edu PloS one 2012 Only protein-protein interactions imex curation imex curation navarro-2012-3 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. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ., Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ. pmccormick@ub.edu PloS one 2012 Only protein-protein interactions imex curation imex curation navarro-2012-5 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. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag visualisation tag visualisation Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ., Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ. BRET with BiFC assay;Sequential Split Renilla Luciferase Complementation followed by Bimolecular Fluorescence Complementation (BIFC). One D2 receptor was fused to the N-terminal fragment (nRluc8) and another D2 receptor was fused to the C-terminal fragment (cRluc8) acting as BRET donor after full length Rluc8 reconstitution; one Sigma receptor was fused to an YFP Venus N-terminal fragment (nVenus) and another Sigma receptor was fused to the YFP Venus C-terminal fragment (cVenus), acting as BRET acceptor after YFP Venus reconstitution. pmccormick@ub.edu PloS one 2012 Only protein-protein interactions imex curation imex curation navarro-2012-6 mouse-brain-striatum Mouse striatum brain_striatum Brain striatum, Neostratum. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ., Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ. pmccormick@ub.edu PloS one 2012 Only protein-protein interactions imex curation imex curation navarro-2012-7 mouse-brain-striatum Mouse striatum brain_striatum Brain striatum, Neostratum. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ., Navarro G., Moreno E., Bonaventura J., Brugarolas M., Farré D., Aguinaga D., Mallol J., Cortés A., Casadó V., Lluís C., Ferre S., Franco R., Canela E., McCormick PJ. pmccormick@ub.edu PloS one 2012 Only protein-protein interactions imex curation imex curation sgmr1_human Sigma non-opioid intracellular receptor 1 SIGMAR1 OPRS1 SRBP Aging-associated gene 8 protein SR31747-binding protein Sigma 1-type opioid receptor AAG8 protein protein human Homo sapiens Human MQWAVGRRWAWAALLLAVAAVLTQVVWLWLGTQSFVFQREEIAQLARQYAGLDHELAFSRLIVELRRLHPGHVLPDEELQWVFVNAGGWMGAMCLLHASLSEYVLLFGTALGSRGHSGRYWAEISDTIISGTFHQWREGTTKSEVFYPGETVVHGPGEATAVEWGPNTWMVEYGRGVIPSTLAFALADTVFSTQDFLTLFYTLRSYARGLRLELTTYLFGQDP drd2_human D(2) dopamine receptor DRD2 Dopamine D2 receptor protein protein human Homo sapiens Human MDPLNLSWYDDDLERQNWSRPFNGSDGKADRPHYNYYATLLTLLIAVIVFGNVLVCMAVSREKALQTTTNYLIVSLAVADLLVATLVMPWVVYLEVVGEWKFSRIHCDIFVTLDVMMCTASILNLCAISIDRYTAVAMPMLYNTRYSSKRRVTVMISIVWVLSFTISCPLLFGLNNADQNECIIANPAFVVYSSIVSFYVPFIVTLLVYIKIYIVLRRRRKRVNTKRSSRAFRAHLRAPLKGNCTHPEDMKLCTVIMKSNGSFPVNRRRVEAARRAQELEMEMLSSTSPPERTRYSPIPPSHHQLTLPDPSHHGLHSTPDSPAKPEKNGHAKDHPKIAKIFEIQTMPNGKTRTSLKTMSRRKLSQQKEKKATQMLAIVLGVFIICWLPFFITHILNIHCDCNIPPVLYSAFTWLGYVNSAVNPIIYTTFNIEFRKAFLKILHC drd2_mouse D(2) dopamine receptor Dopamine D2 receptor Drd2 protein protein mouse Mus musculus Mouse MDPLNLSWYDDDLERQNWSRPFNGSEGKPDRPHYNYYAMLLTLLIFIIVFGNVLVCMAVSREKALQTTTNYLIVSLAVADLLVATLVMPWVVYLEVVGEWKFSRIHCDIFVTLDVMMCTASILNLCAISIDRYTAVAMPMLYNTRYSSKRRVTVMIAIVWVLSFTISCPLLFGLNNTDQNECIIANPAFVVYSSIVSFYVPFIVTLLVYIKIYIVLRKRRKRVNTKRSSRAFRANLKTPLKGNCTHPEDMKLCTVIMKSNGSFPVNRRRMDAARRAQELEMEMLSSTSPPERTRYSPIPPSHHQLTLPDPSHHGLHSNPDSPAKPEKNGHAKIVNPRIAKFFEIQTMPNGKTRTSLKTMSRRKLSQQKEKKATQMLAIVLGVFIICWLPFFITHILNIHCDCNIPPVLYSAFTWLGYVNSAVNPIIYTTFNIEFRKAFMKILHC mint This is one of the five types (D1 to D5) of receptors for dopamine. The activity of this receptor is mediated by G proteins which inhibit adenylyl cyclase sgmr1_mouse Sigma non-opioid intracellular receptor 1 Sigma 1-type opioid receptor Sigmar1 Oprs1 protein protein mouse Mus musculus Mouse MPWAAGRRWAWITLILTIIAVLIQAAWLWLGTQNFVFSREEIAQLARQYAGLDHELAFSRLIVELRRLHPGHVLPDEELQWVFVNAGGWMGAMCILHASLSEYVLLFGTALGSHGHSGRYWAEISDTIISGTFHQWKEGTTKSEVFYPGETVVHGPGEATALEWGPNTWMVEYGRGVIPSTLFFALADTFFSTQDYLTLFYTLRAYARGLRLELTTYLFGQDS drd2-sigmar1-1 3577107 N/A 3577113 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role luminescence donor luminescence donor over-expressed over expressed level over-expressed C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 1A, 3A Cocaine;CHEBI:27958 PRE-084;a sigma receptor agonist, selective for the sigma-1 subtype. drd2-sigmar1-3 3577107 N/A 3577113 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577113 unspecified role unspecified role luminescence donor luminescence donor over-expressed over expressed level over-expressed C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 2A, 3B No significant changes in BRETmax or BRET50 were observed on addition of either agonist PRE084 or antagonist PD144418 or cocaine. drd2-sigmar1-4 3577107 N/A 3577114 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role luminescence donor luminescence donor over-expressed over expressed level over-expressed C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 3C/D Cocaine;CHEBI:27958 PRE-084;a sigma receptor agonist, selective for the sigma-1 subtype. Treatment with Sigma-1 siRNA completely abolished the effect of cocaine or PRE084 on the BRET saturation curve. drd2-sigmar1-5 3577108 N/A 3577113 unspecified role unspecified role luminescence donor luminescence donor over-expressed over expressed level over-expressed C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577113 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role luminescence transmitter luminescence transmitter over-expressed over expressed level over-expressed C-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag 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 Figure 2C drd2-sigmar1-6 3577108 N/A 3577113 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role luminescence donor luminescence donor over-expressed over expressed level over-expressed C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role luminescence transmitter luminescence transmitter over-expressed over expressed level over-expressed C-terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag 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 Figure 2B drd2-sigmar1-2 3577109 N/A 3577113 immunocytochemistry immunocytochemistry unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus renilla luciferase renilla luciferase protein tag renilla luciferase c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Figure 1C drd2-sigmar1-7 3577110 N/A 3577113 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed C-terminus venus-c venus c-terminal fragment c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577113 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed C-terminus venus-n venus n-terminal fragment c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed C-terminus luciferase-n luciferase-n luciferase-n c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3577114 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed C-terminus luciferase-c luciferase-c luciferase-c c-term range C-terminal range c-term range c-term range C-terminal range c-term range false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 2D drd2-sigmar1-8 3577111 N/A 3577115 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3577116 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Figure 6A drd2-sigmar1-9 3577112 N/A 3577115 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3577116 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 6 B/D