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/ Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 imex curation Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. mguzman@quim.ucm.es,clluis@ub.edu 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-brain-striatum HUMAN BRAIN STRIATUM brain_striatum Brain striatum, Neostratum. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. Human caudate-putamen post mortem samples from control subjects and Huntingdon's Disease patients. mguzman@quim.ucm.es,clluis@ub.edu Accepted 2019-OCT-02 AT 10:43 BST AT 10:43 BST by MARTA Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 Only protein-protein interactions imex curation imex curation Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 imex curation Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. mguzman@quim.ucm.es,clluis@ub.edu 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-brain-striatum Mouse striatum brain_striatum Brain striatum, Neostratum. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. Dorsal-striatum sections from:- wild-type HdhQ7/Q7 mice, mutant huntingtin-expressing heterozygous HdhQ7/Q111 mice, mice transgenic for exon 1 of the human huntingtin gene with a greatly expanded CAG tract (R6/1 mice, ~115 CAG repeats; R6/2 mice, ~160 CAG repeats). mguzman@quim.ucm.es,clluis@ub.edu Accepted 2019-OCT-02 AT 10:43 BST AT 10:43 BST by MARTA Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 Only protein-protein interactions imex curation imex curation Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 imex curation Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. mguzman@quim.ucm.es,clluis@ub.edu 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-brain-striatum Mouse striatum brain_striatum Brain striatum, Neostratum. pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. I have not amalgamed with Expt-5 as there are no comparisons with polyGlutamine mutants in this experiment - different Exp-Mod. dorsal striatum from mice of differing genotypes. mguzman@quim.ucm.es,clluis@ub.edu Accepted 2019-OCT-02 AT 10:43 BST AT 10:43 BST by MARTA Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 Only protein-protein interactions imex curation imex curation Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 imex curation Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. mguzman@quim.ucm.es,clluis@ub.edu 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-STHdhQ7/Q7 mouse STHdhQ7/Q7 sthdhq7/q7 STHdhQ7/Q7 mouse (brain)- striatal derived cell line from a knock in transgenic mouse containing homozygous Huntingtin (HTT) loci with a humanized Exon 1 containing 7 polyglutamine repeats. CVCL_M590 pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. Huntingtin-expressing STHdhQ7 striatal neuroblasts. These wild-type cells express Huntingtin with 7 glutamine residues. mguzman@quim.ucm.es,clluis@ub.edu Accepted 2019-OCT-02 AT 10:43 BST AT 10:43 BST by MARTA Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 Only protein-protein interactions imex curation imex curation Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 imex curation Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. mguzman@quim.ucm.es,clluis@ub.edu 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-STHdhq111/q111 immortalised embryonic striatal cells with humanised HTT Exon 1 and 111 CAG repeats sthdhq111 STHdh Q111/Q111 https://www.coriell.org/0/sections/Search/Sample_Detail.aspx?Ref=CH00095&PgId=166 Coriell Cell Repositories;CH00095 Cellosaurus STHdhQ111 (CVCL_M591) pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. Mutant Huntingtin-expressing STHdhQ111 striatal neuroblasts. These cells express mutant Huntingtin with 111 glutamine residues. mguzman@quim.ucm.es,clluis@ub.edu Accepted 2019-OCT-02 AT 10:43 BST AT 10:43 BST by MARTA Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 Only protein-protein interactions imex curation imex curation Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 imex curation Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. mguzman@quim.ucm.es,clluis@ub.edu 21-Nov-2019: 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. bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Moreno E., Chiarlone A., Medrano M., Puigdellívol M., Bibic L., Howell LA., Resel E., Puente N., Casarejos MJ., Perucho J., Botta J., Suelves N., Ciruela F., Ginés S., Galve-Roperh I., Casadó V., Grandes P., Lutz B., Monory K., Canela EI., Lluís C., McCormick PJ., Guzmán M. mguzman@quim.ucm.es,clluis@ub.edu Accepted 2019-OCT-02 AT 10:43 BST AT 10:43 BST by MARTA Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 2018 Only protein-protein interactions imex curation imex curation cnr1_mouse Cannabinoid receptor 1 Cnr1 Brain-type cannabinoid receptor protein protein mouse Mus musculus Mouse MKSILDGLADTTFRTITTDLLYVGSNDIQYEDIKGDMASKLGYFPQKFPLTSFRGSPFQEKMTAGDNSPLVPAGDTTNITEFYNKSLSSFKENEDNIQCGENFMDMECFMILNPSQQLAIAVLSLTLGTFTVLENLLVLCVILHSRSLRCRPSYHFIGSLAVADLLGSVIFVYSFVDFHVFHRKDSPNVFLFKLGGVTASFTASVGSLFLTAIDRYISIHRPLAYKRIVTRPKAVVAFCLMWTIAIVIAVLPLLGWNCKKLQSVCSDIFPLIDETYLMFWIGVTSVLLLFIVYAYMYILWKAHSHAVRMIQRGTQKSIIIHTSEDGKVQVTRPDQARMDIRLAKTLVLILVVLIICWGPLLAIMVYDVFGKMNKLIKTVFAFCSMLCLLNSTVNPIIYALRSKDLRHAFRSMFPSCEGTAQPLDNSMGDSDCLHKHANNTASMHRAAESCIKSTVKIAKVTMSVSTDTSAEAL E504168191CB6429 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 2EC8460CA11FC7F0 aa2ar_mouse Adenosine receptor A2a Adora2a protein protein mouse Mus musculus Mouse MGSSVYIMVELAIAVLAILGNVLVCWAVWINSNLQNVTNFFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFFACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGMRAKGIIAICWVLSFAIGLTPMLGWNNCSQKDENSTKTCGEGRVTCLFEDVVPMNYMVYYNFFAFVLLPLLLMLAIYLRIFLAARRQLKQMESQPLPGERTRSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCSTCQHAPPWLMYLAIILSHSNSVVNPFIYAYRIREFRQTFRKIIRTHVLRRQEPFRAGGSSAWALAAHSTEGEQVSLRLNGHPLGVWANGSAPHSGRRPNGYTLGPGGGGSTQGSPGDVELLTQEHQEGQEHPGLGDHLAQGRVGTASWSSEFAPS E5670F442D243E71 aa2ar_human Adenosine receptor A2a ADORA2A ADORA2 protein protein human Homo sapiens Human MPIMGSSVYITVELAIAVLAILGNVLVCWAVWLNSNLQNVTNYFVVSLAAADIAVGVLAIPFAITISTGFCAACHGCLFIACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGTRAKGIIAICWVLSFAIGLTPMLGWNNCGQPKEGKNHSQGCGEGQVACLFEDVVPMNYMVYFNFFACVLVPLLLMLGVYLRIFLAARRQLKQMESQPLPGERARSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCPDCSHAPLWLMYLAIVLSHTNSVVNPFIYAYRIREFRQTFRKIIRSHVLRQQEPFKAAGTSARVLAAHGSDGEQVSLRLNGHPPGVWANGSAPHPERRPNGYALGLVSGGSAQESQGNTGLPDVELLSHELKGVCPEPPGLDDPLAQDGAGVS 9438E9D64A6BE61B cnr1_human Cannabinoid receptor 1 CNR1 CNR CANN6 protein protein human Homo sapiens Human MKSILDGLADTTFRTITTDLLYVGSNDIQYEDIKGDMASKLGYFPQKFPLTSFRGSPFQEKMTAGDNPQLVPADQVNITEFYNKSLSSFKENEENIQCGENFMDIECFMVLNPSQQLAIAVLSLTLGTFTVLENLLVLCVILHSRSLRCRPSYHFIGSLAVADLLGSVIFVYSFIDFHVFHRKDSRNVFLFKLGGVTASFTASVGSLFLTAIDRYISIHRPLAYKRIVTRPKAVVAFCLMWTIAIVIAVLPLLGWNCEKLQSVCSDIFPHIDETYLMFWIGVTSVLLLFIVYAYMYILWKAHSHAVRMIQRGTQKSIIIHTSEDGKVQVTRPDQARMDIRLAKTLVLILVVLIICWGPLLAIMVYDVFGKMNKLIKTVFAFCSMLCLLNSTVNPIIYALRSKDLRHAFRSMFPSCEGTAQPLDNSMGDSDCLHKHANNAASVHRAAESCIKSTVKIAKVTMSVSTDTSAEAL DIP protein P21554 original sequence version: 162 1D2E49061D12ABF2 adora2a-cnr1-2 3 7 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 9 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 Figure 1, Supplementary Figure 2 When using tissue from mutant mice bearing a genetic deletion of CB1R in forebrain GABAergic neurons, a reduction in the number of dots was observed, but no reduction was seen when using mice with a genetic deletion of CB1R glutamatergic neurons. cnr1-adora2a-1 6 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus cyfp cYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus nyfp nYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 2B adora2a-cnr1-3 6 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus cyfp cYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 competitor competitor competitor unspecified role unspecified role TM5_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain C-terminus tat tag transactivating tag Tat tag tat tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range chemical synthesis Chemical synthesis (Chemical synthesis) 11 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus nyfp nYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 2B-2 adora2a-cnr1-4 6 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus cyfp cYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 competitor competitor competitor unspecified role unspecified role TM6_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminus tat tag transactivating tag Tat tag tat tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range chemical synthesis Chemical synthesis (Chemical synthesis) 11 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed C-terminus nyfp nYFP c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 2B-3 adora2a-cnr1-5 5 7 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 9 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 Figure 3a Expression of mutant huntingtin does not prevent heteromerization of both receptors. adora2a-cnr1-7 2 7 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 9 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 Figure 4A, Supplementary Figure S7a/b By the age of 4 months in heterozygous mutant HdhQ7/Q111 mice expression of A2AR-CB1R heteromers was at similar levels as wild-type, but by the age of 6-8 months was almost completely lost. A similar result was obtained using R6/1 or R6/2 mice. adora2a-cnr1-6 4 7 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 9 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 Figure 3a adora2a-cnr1-8 1 10 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 11 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 Figure 5, Supplementary figure 8 A2AR-CB1R heteromers were readily evident in the caudate-putamen of control individuals, and were also detected at those normal levels in asymptomatic huntingtin gene-mutation carriers (HD grade 0) and early symptomatic HD patients (HD grades 1–2). However number of cells with A2AR-CB1R heteromers was strongly reduced in caudate-putamen samples from high-grade, advanced HD patients (HD grades 3–4). adora2a-cnr1-9 2 8 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level 7 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 Supplementary Figure 6C, 7C CB1R-D2R heteromer expression was not reduced in 6-month-old mutant HdhQ7/Q111 mice compared with their wild-type controls. A similar result was obtained using R6/1 or R6/2 mice.