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/ Dopamine D2 and D4 receptor heteromerization and its allosteric receptor-receptor interactions. Dopamine D2 and D4 receptor heteromerization and its allosteric receptor-receptor interactions. Biochemical and biophysical research communications 2011 imex curation Borroto-Escuela DO., Van Craenenbroeck K., Romero-Fernandez W., Guidolin D., Woods AS., Rivera A., Haegeman G., Agnati LF., Tarakanov AO., Fuxe K. Dark space Dataset = no 10? Kjell.Fuxe@ki.se 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Borroto-Escuela DO., Van Craenenbroeck K., Romero-Fernandez W., Guidolin D., Woods AS., Rivera A., Haegeman G., Agnati LF., Tarakanov AO., Fuxe K. Dark space Dataset = no 10? Kjell.Fuxe@ki.se Accepted 2016-APR-28 AT 09:44 BST AT 09:44 BST by ORCHARD Biochemical and biophysical research communications 2011 Only protein-protein interactions imex curation imex curation Dopamine D2 and D4 receptor heteromerization and its allosteric receptor-receptor interactions. Dopamine D2 and D4 receptor heteromerization and its allosteric receptor-receptor interactions. Biochemical and biophysical research communications 2011 imex curation Borroto-Escuela DO., Van Craenenbroeck K., Romero-Fernandez W., Guidolin D., Woods AS., Rivera A., Haegeman G., Agnati LF., Tarakanov AO., Fuxe K. Dark space Dataset = no 10? Kjell.Fuxe@ki.se 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Borroto-Escuela DO., Van Craenenbroeck K., Romero-Fernandez W., Guidolin D., Woods AS., Rivera A., Haegeman G., Agnati LF., Tarakanov AO., Fuxe K. Dark space Dataset = no 10? Kjell.Fuxe@ki.se Accepted 2016-APR-28 AT 09:44 BST AT 09:44 BST by ORCHARD Biochemical and biophysical research communications 2011 Only protein-protein interactions imex curation imex curation Dopamine D2 and D4 receptor heteromerization and its allosteric receptor-receptor interactions. Dopamine D2 and D4 receptor heteromerization and its allosteric receptor-receptor interactions. Biochemical and biophysical research communications 2011 imex curation Borroto-Escuela DO., Van Craenenbroeck K., Romero-Fernandez W., Guidolin D., Woods AS., Rivera A., Haegeman G., Agnati LF., Tarakanov AO., Fuxe K. Dark space Dataset = no 10? Kjell.Fuxe@ki.se 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Borroto-Escuela DO., Van Craenenbroeck K., Romero-Fernandez W., Guidolin D., Woods AS., Rivera A., Haegeman G., Agnati LF., Tarakanov AO., Fuxe K. In Supp fig 2 I am not 100% sure what they meant by "wild-type receptors" in this context. (no description I can find in Methods section) I think they must mean adding ?untagged D2- long, OR D4.4 to the heterodimer? Dark space Dataset = no 10? Kjell.Fuxe@ki.se Accepted 2016-APR-28 AT 09:44 BST AT 09:44 BST by ORCHARD Biochemical and biophysical research communications 2011 Only protein-protein interactions imex curation imex curation drd4_human D(4) dopamine receptor DRD4 D(2C) dopamine receptor Dopamine D4 receptor protein protein human Homo sapiens Human MGNRSTADADGLLAGRGPAAGASAGASAGLAGQGAAALVGGVLLIGAVLAGNSLVCVSVATERALQTPTNSFIVSLAAADLLLALLVLPLFVYSEVQGGAWLLSPRLCDALMAMDVMLCTASIFNLCAISVDRFVAVAVPLRYNRQGGSRRQLLLIGATWLLSAAVAAPVLCGLNDVRGRDPAVCRLEDRDYVVYSSVCSFFLPCPLMLLLYWATFRGLQRWEVARRAKLHGRAPRRPSGPGPPSPTPPAPRLPQDPCGPDCAPPAPGLPRGPCGPDCAPAAPSLPQDPCGPDCAPPAPGLPPDPCGSNCAPPDAVRAAALPPQTPPQTRRRRRAKITGRERKAMRVLPVVVGAFLLCWTPFFVVHITQALCPACSVPPRLVSAVTWLGYVNSALNPVIYTVFNAEFRNVFRKALRACC DIP protein P21917 original sequence version: 120 mint 34CBE8320302008E p14416-1 D(2) dopamine receptor Dopamine D2 receptor D2(Long) DRD2 D2A protein protein human Homo sapiens Human MDPLNLSWYDDDLERQNWSRPFNGSDGKADRPHYNYYATLLTLLIAVIVFGNVLVCMAVSREKALQTTTNYLIVSLAVADLLVATLVMPWVVYLEVVGEWKFSRIHCDIFVTLDVMMCTASILNLCAISIDRYTAVAMPMLYNTRYSSKRRVTVMISIVWVLSFTISCPLLFGLNNADQNECIIANPAFVVYSSIVSFYVPFIVTLLVYIKIYIVLRRRRKRVNTKRSSRAFRAHLRAPLKGNCTHPEDMKLCTVIMKSNGSFPVNRRRVEAARRAQELEMEMLSSTSPPERTRYSPIPPSHHQLTLPDPSHHGLHSTPDSPAKPEKNGHAKDHPKIAKIFEIQTMPNGKTRTSLKTMSRRKLSQQKEKKATQMLAIVLGVFIICWLPFFITHILNIHCDCNIPPVLYSAFTWLGYVNSAVNPIIYTTFNIEFRKAFLKILHC 9BF8EA36C988A2E2 p14416-2 D(2) dopamine receptor Dopamine D2 receptor D2(Short) DRD2 D2B protein protein human Homo sapiens Human MDPLNLSWYDDDLERQNWSRPFNGSDGKADRPHYNYYATLLTLLIAVIVFGNVLVCMAVSREKALQTTTNYLIVSLAVADLLVATLVMPWVVYLEVVGEWKFSRIHCDIFVTLDVMMCTASILNLCAISIDRYTAVAMPMLYNTRYSSKRRVTVMISIVWVLSFTISCPLLFGLNNADQNECIIANPAFVVYSSIVSFYVPFIVTLLVYIKIYIVLRRRRKRVNTKRSSRAFRAHLRAPLKEAARRAQELEMEMLSSTSPPERTRYSPIPPSHHQLTLPDPSHHGLHSTPDSPAKPEKNGHAKDHPKIAKIFEIQTMPNGKTRTSLKTMSRRKLSQQKEKKATQMLAIVLGVFIICWLPFFITHILNIHCDCNIPPVLYSAFTWLGYVNSAVNPIIYTTFNIEFRKAFLKILHC BFC25AC601DFDC7F drd2-drd4-1 1 4 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 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 1A The number of repeats of 16 amino acids in the third cytoplasmic loop is highly polymorphic and varies among different alleles. Alleles corresponding in size to a 2 (D4.2), 3 (D4.3), 4 (D4.4), 5 (D4.5), 6 (D4.6), 7 (D4.7) and 9 (D4.9) repeats have been described. All variants (4.2, 4.4 4.7) of DRD4 interacted with DRD2 short form. drd2-drd4-2 2 4 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction 1C The number of repeats of 16 amino acids in the third cytoplasmic loop is highly polymorphic and varies among different alleles. Alleles corresponding in size to a 2 (D4.2), 3 (D4.3), 4 (D4.4), 5 (D4.5), 6 (D4.6), 7 (D4.7) and 9 (D4.9) repeats have been described. All variants of DRD4 tested (4.2, 4.4, 4.7) interacted with DRD2 long form, the lowest number of red clusters being seen for the longest variant of DRD4 (DRD4-7). drd2-drd4-3 3 4 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 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 5 unspecified role unspecified role fluorescence donor fluorescence donor c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range proximity proximity close-contact colocalization neighbourhood interaction 2 All variants of DRD4 interacted with DRD2 long form. The DRD4 longest (4-7) variant shows lesser affinity, with reduced BRETmax and increased BRET50 values, compared to other variants. drd2-drd4-4 3 4 competitor competitor competitor unspecified role unspecified role 4 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 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 5 unspecified role unspecified role fluorescence donor fluorescence donor c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range proximity proximity close-contact colocalization neighbourhood interaction Supp fig 2 All variants of DRD4 interacted with DRD2 long form. The DRD4 longest (4-7) variant shows lesser affinity, with reduced BRETmax and increased BRET50 values, compared to other variants. drd2-drd4-5 3 4 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 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 5 competitor competitor competitor unspecified role unspecified role 5 unspecified role unspecified role fluorescence donor fluorescence donor c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range proximity proximity close-contact colocalization neighbourhood interaction Supp fig 2i All variants of DRD4 interacted with DRD2 long form. The DRD4 longest (4-7) variant shows lesser affinity, with reduced BRETmax and increased BRET50 values, compared to other variants. drd2-drd4-6 3 4 unspecified role unspecified role fluorescence donor fluorescence donor region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 4 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain 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 proximity proximity close-contact colocalization neighbourhood interaction Supplementary fig 3 With regard to homomer formation, D4.7R showed the greatest affinity. drd2-drd4-7 3 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept 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 5 unspecified role unspecified role fluorescence donor fluorescence donor c_terminus yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range proximity proximity close-contact colocalization neighbourhood interaction Supp figure 3