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/ Neuronal Calcium and cAMP Cross-Talk Mediated by Cannabinoid CB1 Receptor and EF-Hand Calcium Sensor Interactions. Neuronal Calcium and cAMP Cross-Talk Mediated by Cannabinoid CB1 Receptor and EF-Hand Calcium Sensor Interactions. Frontiers in Cell and Developmental Biology 2017 imex curation Angelats E., Requesens M., Aguinaga D., Kreutz MR., Franco R., Navarro G. Interactions involving a Calneuron-1 construct truncated at the C terminus (calneuron-1ΔC) and an N-terminal myristoylation-deficient NCS1 mutant have not been included as no details of these construts can be traced and the authors did not reply to a request for information. rfranco123@gmail.com,rfranco@ub.edu 03-Oct-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. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Angelats E., Requesens M., Aguinaga D., Kreutz MR., Franco R., Navarro G. Interactions involving a Calneuron-1 construct truncated at the C terminus (calneuron-1ΔC) and an N-terminal myristoylation-deficient NCS1 mutant have not been included as no details of these construts can be traced and the authors did not reply to a request for information. rfranco123@gmail.com,rfranco@ub.edu Accepted 2019-SEP-27 AT 13:53 BST AT 13:53 BST by MARTA Frontiers in Cell and Developmental Biology 2017 Only protein-protein interactions imex curation imex curation Neuronal Calcium and cAMP Cross-Talk Mediated by Cannabinoid CB1 Receptor and EF-Hand Calcium Sensor Interactions. Neuronal Calcium and cAMP Cross-Talk Mediated by Cannabinoid CB1 Receptor and EF-Hand Calcium Sensor Interactions. Frontiers in Cell and Developmental Biology 2017 imex curation Angelats E., Requesens M., Aguinaga D., Kreutz MR., Franco R., Navarro G. Interactions involving a Calneuron-1 construct truncated at the C terminus (calneuron-1ΔC) and an N-terminal myristoylation-deficient NCS1 mutant have not been included as no details of these construts can be traced and the authors did not reply to a request for information. rfranco123@gmail.com,rfranco@ub.edu 03-Oct-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Angelats E., Requesens M., Aguinaga D., Kreutz MR., Franco R., Navarro G. mouse striatal neurons. Interactions involving a Calneuron-1 construct truncated at the C terminus (calneuron-1ΔC) and an N-terminal myristoylation-deficient NCS1 mutant have not been included as no details of these construts can be traced and the authors did not reply to a request for information. rfranco123@gmail.com,rfranco@ub.edu Accepted 2019-SEP-27 AT 13:53 BST AT 13:53 BST by MARTA Frontiers in Cell and Developmental Biology 2017 Only protein-protein interactions imex curation imex curation Neuronal Calcium and cAMP Cross-Talk Mediated by Cannabinoid CB1 Receptor and EF-Hand Calcium Sensor Interactions. Neuronal Calcium and cAMP Cross-Talk Mediated by Cannabinoid CB1 Receptor and EF-Hand Calcium Sensor Interactions. Frontiers in Cell and Developmental Biology 2017 imex curation Angelats E., Requesens M., Aguinaga D., Kreutz MR., Franco R., Navarro G. Interactions involving a Calneuron-1 construct truncated at the C terminus (calneuron-1ΔC) and an N-terminal myristoylation-deficient NCS1 mutant have not been included as no details of these construts can be traced and the authors did not reply to a request for information. rfranco123@gmail.com,rfranco@ub.edu 03-Oct-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. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag visualisation tag visualisation Angelats E., Requesens M., Aguinaga D., Kreutz MR., Franco R., Navarro G. Interactions involving the N-terminal myristoylation-deficient NCS1 mutant and/or the calneuron 1 c-terminally deleted mutant have not been included as insufficient details of these mutants can be traced. Fig 2A:the calneuron 1 c-terminally deleted mutant (n-terminal YFP) as stated in pmid: 19682431 -ref 13 from the stated Hradsky-2013 reference, (also Hradsky 2011 Ref.) is of RATTINE origin. As it did not bind in Fig 2A = no interaction and as not Human so cannot add a "required to bind" feature..Similarly for the NCS1 mutant. Interactions involving a Calneuron-1 construct truncated at the C terminus (calneuron-1ΔC) and an N-terminal myristoylation-deficient NCS1 mutant have not been included as no details of these construts can be traced and the authors did not reply to a request for information. rfranco123@gmail.com,rfranco@ub.edu Accepted 2019-SEP-27 AT 13:53 BST AT 13:53 BST by MARTA Frontiers in Cell and Developmental Biology 2017 Only protein-protein interactions imex curation imex curation cabp8_mouse Calcium-binding protein 8 Calneuron I Calneuron-1 MNCb-0849 Caln1 Cabp8 protein protein mouse Mus musculus Mouse MRLPEQPGDGKPENETKGDQETPERGEEPRRSPAPDFPTWEKMPFHHVTAGLLYKGNYLNRSLSAGSDSEQLANISVEELDEIREAFRVLDRDGNGFISKQELGMAMRSLGYMPSEVELAIIMQRLDMDGDGQVDFDEFMTILGPKLVSSEGRDGFLGNTIDSIFWQFDMQRVTLEELKHILYHAFRDHLTMKDIENIIINEEESLNETSGNCQTEFEGVHSQKQNRQTCVRKSLICAFAMAFIISVMLIAANQILRSGME D38AE0DFBBAE38D9 cnr1_human Cannabinoid receptor 1 CNR1 CNR CANN6 protein protein human Homo sapiens Human MKSILDGLADTTFRTITTDLLYVGSNDIQYEDIKGDMASKLGYFPQKFPLTSFRGSPFQEKMTAGDNPQLVPADQVNITEFYNKSLSSFKENEENIQCGENFMDIECFMVLNPSQQLAIAVLSLTLGTFTVLENLLVLCVILHSRSLRCRPSYHFIGSLAVADLLGSVIFVYSFIDFHVFHRKDSRNVFLFKLGGVTASFTASVGSLFLTAIDRYISIHRPLAYKRIVTRPKAVVAFCLMWTIAIVIAVLPLLGWNCEKLQSVCSDIFPHIDETYLMFWIGVTSVLLLFIVYAYMYILWKAHSHAVRMIQRGTQKSIIIHTSEDGKVQVTRPDQARMDIRLAKTLVLILVVLIICWGPLLAIMVYDVFGKMNKLIKTVFAFCSMLCLLNSTVNPIIYALRSKDLRHAFRSMFPSCEGTAQPLDNSMGDSDCLHKHANNAASVHRAAESCIKSTVKIAKVTMSVSTDTSAEAL DIP protein P21554 original sequence version: 162 1D2E49061D12ABF2 ncs1_mouse Neuronal calcium sensor 1 Ncs1 Freq Frequenin homolog protein protein mouse Mus musculus Mouse MGKSNSKLKPEVVEELTRKTYFTEKEVQQWYKGFIKDCPSGQLDAAGFQKIYKQFFPFGDPTKFATFVFNVFDENKDGRIEFSEFIQALSVTSRGTLDEKLRWAFKLYDLDNDGYITRNEMLDIVDAIYQMVGNTVELPEEENTPEKRVDRIFAMMDKNADGKLTLQEFQEGSKADPSIVQALSLYDGLV DIP protein Q8BNY6 original sequence version: 99 Neuronal calcium sensor, regulator of G protein-coupled receptor phosphorylation in a calcium dependent manner. Directly regulates GRK1 (RHOK), but not GRK2 to GRK5. Can substitute for calmodulin mint 9AF8E26A23F80D4F cnr1_mouse Cannabinoid receptor 1 Cnr1 Brain-type cannabinoid receptor protein protein mouse Mus musculus Mouse MKSILDGLADTTFRTITTDLLYVGSNDIQYEDIKGDMASKLGYFPQKFPLTSFRGSPFQEKMTAGDNSPLVPAGDTTNITEFYNKSLSSFKENEDNIQCGENFMDMECFMILNPSQQLAIAVLSLTLGTFTVLENLLVLCVILHSRSLRCRPSYHFIGSLAVADLLGSVIFVYSFVDFHVFHRKDSPNVFLFKLGGVTASFTASVGSLFLTAIDRYISIHRPLAYKRIVTRPKAVVAFCLMWTIAIVIAVLPLLGWNCKKLQSVCSDIFPLIDETYLMFWIGVTSVLLLFIVYAYMYILWKAHSHAVRMIQRGTQKSIIIHTSEDGKVQVTRPDQARMDIRLAKTLVLILVVLIICWGPLLAIMVYDVFGKMNKLIKTVFAFCSMLCLLNSTVNPIIYALRSKDLRHAFRSMFPSCEGTAQPLDNSMGDSDCLHKHANNTASMHRAAESCIKSTVKIAKVTMSVSTDTSAEAL E504168191CB6429 cabp8_human Calcium-binding protein 8 CALN1 CABP8 Calneuron I Calneuron-1 protein protein human Homo sapiens Human MRLPEQPGEGKPENEKKGDGGALGGGEEPPRSQAPDFPTWEKMPFHHVTAGLLYKGNYLNRSLSAGSDSEQLANISVEELDEIREAFRVLDRDGNGFISKQELGMAMRSLGYMPSEVELAIIMQRLDMDGDGQVDFDEFMTILGPKLVSSEGRDGFLGNTIDSIFWQFDMQRITLEELKHILYHAFRDHLTMKDIENIIINEEESLNETSGNCQTEFEGVHSQKQNRQTCVRKSLICAFAMAFIISVMLIAANQILRSGME 8276320AD714FACD ncs1_human Neuronal calcium sensor 1 Frequenin homolog Frequenin-like protein Frequenin-like ubiquitous protein NCS1 FLUP FREQ protein protein human Homo sapiens Human MGKSNSKLKPEVVEELTRKTYFTEKEVQQWYKGFIKDCPSGQLDAAGFQKIYKQFFPFGDPTKFATFVFNVFDENKDGRIEFSEFIQALSVTSRGTLDEKLRWAFKLYDLDNDGYITRNEMLDIVDAIYQMVGNTVELPEEENTPEKRVDRIFAMMDKNADGKLTLQEFQEGSKADPSIVQALSLYDGLV 9AF8E26A23F80D4F caln1-cnr1-1 1 8 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed 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 5 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 colocalization colocalization Figure 1A cnr1-ncs1-1 1 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed 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 5 anti tag immunost anti tag immunostaining anti tag immunost tag fluorescence tag visualisation by fluorescence tag fluorescence 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 colocalization colocalization Figure 1A caln1-cnr1-2 3 8 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed 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 5 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 1B, (2A), 3A A Calneuron-1 construct, truncated at the C terminus and of unknown species origin did not bind Cb1. cnr1-ncs1-2 3 9 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed 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 5 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 1C, (2D), 3B An N-terminal myristoylation-deficient NCS1 mutant of unknown species origin and sequence reduced binding. cnr1-caln1-1 3 8 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 5 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed 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 region sufficient to bind sufficient binding region sufficient to bind range range range range P21554:p.Thr321_Ser322delinsAlaAla mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain TS AA Tue Jun 30 19:19:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr321_Ser322delinsAlaAla' proximity proximity close-contact colocalization neighbourhood interaction Figure 2B/C cnr1-caln1-2 3 9 competitor competitor competitor unspecified role unspecified role over-expressed over expressed level over-expressed 8 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed 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 5 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 3A Increasing intracellular Ca2+ concentrations reduced the competitive effect of of NCS1. not sure if it is OK to add NCS1 as an interactor, but this seemed clearest way to show the interaction + calcium. cnr1-ncs1-3 3 8 competitor competitor competitor unspecified role unspecified role over-expressed over expressed level over-expressed 9 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed 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 5 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 3B again not sure if it is OK to add Caln1 as an interactor, but this seemed clearest way to show the interaction + calcium. Increasing intracellular Ca2+ concentrations did not reduce the competitive effect of of calneuron-1. caln1-cnr1-3 2 4 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 Figure 4A Ionomycin -an ionophore used to raise the intracellular level of calcium (Ca2+). caln1-cnr1-4 2 6 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 Figure 4B Ionomycin -an ionophore used to raise the intracellular level of calcium (Ca2+).