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/ A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. The Journal of biological chemistry 2016 imex curation Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) 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. fluorescence fluorescence technology fluorescence tag fluorescence tag visualisation by fluorescence tag fluorescence Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. The image and image text describe the interactions for SRET as annotated here. However the main text of the publication describes the interactions as the other way around - notably Fig 6C with GHS-R1a-YFP (rather than GSH-R1b-YFP) being added to GHS-R1b-Rluc and GHS-R1a-GFP;similarly for Fig. 6D. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. SRET:Sequential BRET-FRET;BRET combined with FRET. Oxidation of a Renilla Luciferase substrate triggers acceptor excitation by BRET and subsequent energy transfer to a FRET acceptor. YFP emission was determined after adding DeepBlueC as luciferase substrate. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) Accepted 2019-SEP-30 AT 11:27 BST AT 11:27 BST by PPORRAS The Journal of biological chemistry 2016 Only protein-protein interactions imex curation imex curation A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. The Journal of biological chemistry 2016 imex curation Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) 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 Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) Accepted 2019-SEP-30 AT 11:27 BST AT 11:27 BST by PPORRAS The Journal of biological chemistry 2016 Only protein-protein interactions imex curation imex curation A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. The Journal of biological chemistry 2016 imex curation Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) 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 Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) Accepted 2019-SEP-30 AT 11:27 BST AT 11:27 BST by PPORRAS The Journal of biological chemistry 2016 Only protein-protein interactions imex curation imex curation A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. A Significant Role of the Truncated Ghrelin Receptor GHS-R1b in Ghrelin-induced Signaling in Neurons. The Journal of biological chemistry 2016 imex curation Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) 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. copa-ret complemented donor-acceptor resonance energy transfer bret complementation assay bret complementation copa-ret tag fluorescence tag visualisation by fluorescence tag fluorescence Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S., Navarro G., Aguinaga D., Angelats E., Medrano M., Moreno E., Mallol J., Cortés A., Canela EI., Casadó V., McCormick PJ., Lluís C., Ferré S. beta-arrestin-2 recruitment assays have not been annotated as the species origin of arrestin-2 cannot be traced. there are several refs to beta arrestin-2 mainly from Ref 5;none of the mutations given match any species. BRET/BIFC; BRET sensors, the donor Rluc8 and the acceptor YFP Venus, are split into two hemiproteins, with each split sensor being fused to one of the four putative interacting receptors. BRET indicates reconstitution of both sensors and close proximity of the four receptors. sferre@intra.nida.nih.gov,dimartts@hotmail.com The Navarro email address either gets no response or is not active. (dimartts@hotmail.com) Accepted 2019-SEP-30 AT 11:27 BST AT 11:27 BST by PPORRAS The Journal of biological chemistry 2016 Only protein-protein interactions imex curation imex curation drd1_human D(1A) dopamine receptor DRD1 Dopamine D1 receptor protein protein human Homo sapiens Human MRTLNTSAMDGTGLVVERDFSVRILTACFLSLLILSTLLGNTLVCAAVIRFRHLRSKVTNFFVISLAVSDLLVAVLVMPWKAVAEIAGFWPFGSFCNIWVAFDIMCSTASILNLCVISVDRYWAISSPFRYERKMTPKAAFILISVAWTLSVLISFIPVQLSWHKAKPTSPSDGNATSLAETIDNCDSSLSRTYAISSSVISFYIPVAIMIVTYTRIYRIAQKQIRRIAALERAAVHAKNCQTTTGNGKPVECSQPESSFKMSFKRETKVLKTLSVIMGVFVCCWLPFFILNCILPFCGSGETQPFCIDSNTFDVFVWFGWANSSLNPIIYAFNADFRKAFSTLLGCYRLCPATNNAIETVSINNNGAAMFSSHHEPRGSISKECNLVYLIPHAVGSSEDLKKEEAAGIARPLEKLSPALSVILDYDTDVSLEKIQPITQNGQHPT 64E062D765D0DBA7 q92847-2 Growth hormone secretagogue receptor type 1 GH-releasing peptide receptor Ghrelin receptor GHSR protein protein human Homo sapiens Human MWNATPSEEPGFNLTLADLDWDASPGNDSLGDELLQLFPAPLLAGVTATCVALFVVGIAGNLLTMLVVSRFRELRTTTNLYLSSMAFSDLLIFLCMPLDLVRLWQYRPWNFGDLLCKLFQFVSESCTYATVLTITALSVERYFAICFPLRAKVVVTKGRVKLVIFVIWAVAFCSAGPIFVLVGVEHENGTDPWDTNECRPTEFAVRSGLLTVMVWVSSIFFFLPVFCLTVLYSLIGRKLWRRRRGDAVVGASLRDQNHKQTVKMLGGSQRALRLSLAGPILSLCLLPSL 8032FDB4C6BDD42B q92847-1 Growth hormone secretagogue receptor type 1 GH-releasing peptide receptor Ghrelin receptor GHSR protein protein human Homo sapiens Human MWNATPSEEPGFNLTLADLDWDASPGNDSLGDELLQLFPAPLLAGVTATCVALFVVGIAGNLLTMLVVSRFRELRTTTNLYLSSMAFSDLLIFLCMPLDLVRLWQYRPWNFGDLLCKLFQFVSESCTYATVLTITALSVERYFAICFPLRAKVVVTKGRVKLVIFVIWAVAFCSAGPIFVLVGVEHENGTDPWDTNECRPTEFAVRSGLLTVMVWVSSIFFFLPVFCLTVLYSLIGRKLWRRRRGDAVVGASLRDQNHKQTVKMLAVVVFAFILCWLPFHVGRYLFSKSFEPGSLEIAQISQYCNLVSFVLFYLSAAINPILYNIMSKKYRVAVFRLLGFEPFSQRKLSTLKDESSRAWTESSINT D1B62710DA9DC0C6 ghsr-ghsr-11 3 6 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 7 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 colocalization colocalization Figure 1C ghsr-ghsr-12 2 7 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 7 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 6A ghsr-ghsr-13 2 6 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 6 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 6B ghsr-ghsr-14 1 6 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence 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 6 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 7 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 physical association physical association Figure 6C ghsr-ghsr-15 1 7 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence 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 6 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 7 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 physical association physical association Figure 6D ghsr-ghsr-16 4 6 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept 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 7 unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept 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 6 unspecified role unspecified role luminescence donor luminescence donor over-expressed over expressed level over-expressed C-terminus renilla-n renilla-n c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role luminescence donor luminescence donor over-expressed over expressed level over-expressed C-terminus renilla-c renilla-c c-term range C-terminal range c-term range c-term range C-terminal range c-term range 41 39 45 43 physical association physical association Figure 6E drd1-ghsr-1 2 7 unspecified role unspecified role ancillary ancillary over-expressed over expressed level over-expressed 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 6 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 proximity proximity close-contact colocalization neighbourhood interaction Figure 11E In the absence of GHS-R1a cDNA only a non-specific interaction was observed. ghrelin ghrelin plus SKF81297 (a D1R agonist) drd1-ghsr-3 2 7 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 6 unspecified role unspecified role ancillary ancillary over-expressed over expressed level over-expressed proximity proximity close-contact colocalization neighbourhood interaction Figure 11F In the absence of GHS-R1b cDNA only a non-specific interaction was observed.