I2D I2D navarro-2009-2 Interactions between calmodulin, adenosine A2A, and dopamine D2 receptors. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag fluorescence tag visualisation by fluorescence tag fluorescence Navarro G., Aymerich MS., Marcellino D., Cortes A., Casado V., Mallol J., Canela EI., Agnati L., Woods AS., Fuxe K., Lluis C., Lanciego JL., Ferre S., Franco R. rfranco@unav.es J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation Curated by: I2D - Interologous Interaction Database navarro-2009-3 Interactions between calmodulin, adenosine A2A, and dopamine D2 receptors. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Navarro G., Aymerich MS., Marcellino D., Cortes A., Casado V., Mallol J., Canela EI., Agnati L., Woods AS., Fuxe K., Lluis C., Lanciego JL., Ferre S., Franco R. rfranco@unav.es J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation Curated by: I2D - Interologous Interaction Database navarro-2009-1 Interactions between calmodulin, adenosine A2A, and dopamine D2 receptors. rat-brain-striatum Rattus norvegicus brian stiatum brain_striatum Brain striatum, Neostratum. anti bait coip anti bait coimmunoprecipitation anti bait coip sequence tag sequence tag identification sequence tag Navarro G., Aymerich MS., Marcellino D., Cortes A., Casado V., Mallol J., Canela EI., Agnati L., Woods AS., Fuxe K., Lluis C., Lanciego JL., Ferre S., Franco R. rfranco@unav.es J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation Curated by: I2D - Interologous Interaction Database navarro-2009-4 Interactions between calmodulin, adenosine A2A, and dopamine D2 receptors. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Navarro G., Aymerich MS., Marcellino D., Cortes A., Casado V., Mallol J., Canela EI., Agnati L., Woods AS., Fuxe K., Lluis C., Lanciego JL., Ferre S., Franco R. Sequential BRET-FRET (SRET) permits identification of heteromers formed by three different proteins. In SRET, the oxidation of a Renilla luciferase (Rluc) substrate by an Rluc fusion protein triggers acceptor excitation of a second fusion protein by BRET and subsequent FRET to a third fusion protein. rfranco@unav.es J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation Curated by: I2D - Interologous Interaction Database calm_human Calmodulin CALM1 CALM CAM CAM1 CALM2 CAMB CALM3 CAM3 CAMC CAMIII CAM2 CALML2 protein protein human Homo sapiens Human MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK aa2ar_human Adenosine receptor A2a ADORA2A ADORA2 protein protein human Homo sapiens Human MPIMGSSVYITVELAIAVLAILGNVLVCWAVWLNSNLQNVTNYFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFIACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGTRAKGIIAICWVLSFAIGLTPMLGWNNCGQPKEGKNHSQGCGEGQVACLFEDVVPMNYMVYFNFFACVLVPLLLMLGVYLRIFLAARRQLKQMESQPLPGERARSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCPDCSHAPLWLMYLAIVLSHTNSVVNPFIYAYRIREFRQTFRKIIRSHVLRQQEPFKAAGTSARVLAAHGSDGEQVSLRLNGHPPGVWANGSAPHPERRPNGYALGLVSGGSAQESQGNTGLPDVELLSHELKGVCPEPPGLDDPLAQDGAGVS drd2_human D(2) dopamine receptor DRD2 Dopamine D2 receptor protein protein human Homo sapiens Human MDPLNLSWYDDDLERQNWSRPFNGSDGKADRPHYNYYATLLTLLIAVIVFGNVLVCMAVSREKALQTTTNYLIVSLAVADLLVATLVMPWVVYLEVVGEWKFSRIHCDIFVTLDVMMCTASILNLCAISIDRYTAVAMPMLYNTRYSSKRRVTVMISIVWVLSFTISCPLLFGLNNADQNECIIANPAFVVYSSIVSFYVPFIVTLLVYIKIYIVLRRRRKRVNTKRSSRAFRAHLRAPLKGNCTHPEDMKLCTVIMKSNGSFPVNRRRVEAARRAQELEMEMLSSTSPPERTRYSPIPPSHHQLTLPDPSHHGLHSTPDSPAKPEKNGHAKDHPKIAKIFEIQTMPNGKTRTSLKTMSRRKLSQQKEKKATQMLAIVLGVFIICWLPFFITHILNIHCDCNIPPVLYSAFTWLGYVNSAVNPIIYTTFNIEFRKAFLKILHC calm_rat Calmodulin Calm1 Calm Cam Cam1 Calm2 Cam2 Camb Calm3 Cam3 Camc CaMII protein protein rat Rattus norvegicus (Rat) Rat MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK aa2ar_rat Adenosine receptor A2a Adora2a protein protein rat Rattus norvegicus (Rat) Rat MGSSVYITVELAIAVLAILGNVLVCWAVWINSNLQNVTNFFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFFACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGVRAKGIIAICWVLSFAIGLTPMLGWNNCSQKDGNSTKTCGEGRVTCLFEDVVPMNYMVYYNFFAFVLLPLLLMLAIYLRIFLAARRQLKQMESQPLPGERTRSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCSTCRHAPPWLMYLAIILSHSNSVVNPFIYAYRIREFRQTFRKIIRTHVLRRQEPFQAGGSSAWALAAHSTEGEQVSLRLNGHPLGVWANGSATHSGRRPNGYTLGLGGGGSAQGSPRDVELPTQERQEGQEHPGLRGHLVQARVGASSWSSEFAPS calm-a2ar-1 984002 n/a 984006 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 984007 unspecified role unspecified role fluorescence donor fluorescence donor region renilla luciferase renilla luciferase protein tag renilla luciferase undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region required to bind necessary binding region required to bind mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P29274:p.[Arg291Ala;Arg296Ala;Arg300Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false proximity proximity close-contact colocalization neighbourhood interaction false false false Fig. 1a, 2b, 4a Addition of ionomycin for 2 or 5 mins decreases BRET measurement demonstrating that Ca2+ is not required for interaction of Calmodulin with A2aR. Peptide A(291)IREFA(296)QTFA(300) fails to bind calmodulin in Fig. 2b. calm-drd2 984002 n/a 984006 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 984008 unspecified role unspecified role fluorescence donor fluorescence donor region renilla luciferase renilla luciferase protein tag renilla luciferase undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false proximity proximity close-contact colocalization neighbourhood interaction false false false Fig. 1b, 4b Addition of ionomycin for 2, 5 or 10 mins decreases BRET measurement demonstrating that Ca2+ is not required for interaction of Calmodulin with D2 dopamine receptor.. drd2-a2ar 984002 n/a 984007 unspecified role unspecified role fluorescence donor fluorescence donor region renilla luciferase renilla luciferase protein tag renilla luciferase undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 984008 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false proximity proximity close-contact colocalization neighbourhood interaction false false false Fig. 7a Addition of ionomycin for 2 and 5 minutes decreases BRET measurements but increases after 10 minutes incubation. This suggests that Ca2+ binding to CaM induces conformational changes in A2aR but does not disrupt A2aR-DRD2 heteromers. calm-a2ar-2 984003 n/a 984006 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 984007 unspecified role unspecified role neutral component neutral component region renilla luciferase renilla luciferase protein tag renilla luciferase undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 3b,f,j calm-drd2-1 984003 n/a 984006 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 984008 unspecified role unspecified role neutral component neutral component region renilla luciferase renilla luciferase protein tag renilla luciferase undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 3c,g,k calm-a2ar 984004 n/a 984009 unspecified role unspecified role prey prey region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain false region identified peptide identified peptide certain certain sequence position certain certain certain sequence position certain false n/a 984010 unspecified role unspecified role bait bait physical association physical association false false false Table I calm-a2ar-3 984005 n/a 984007 unspecified role unspecified role fluorescence donor fluorescence donor region renilla luciferase renilla luciferase protein tag renilla luciferase undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 984006 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept 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 n/a 984008 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false proximity proximity close-contact colocalization neighbourhood interaction false false false Fig. 6, 7b Addition of 1uM ionomycin in HBSS buffer containing 1.26mM CaCl2 for 10 mins decreases SRET measurement demonstrating that Ca2+ is not required for interaction of Calmodulin-A2aR-DRD2 oligomers.