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/ http://www.ebi.ac.uk/ aguinaga-2019-3 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 Aguinaga D., Medrano M., Cordomí A., Jiménez-Rosés M., Angelats E., Casanovas M., Vega-Quiroga I., Canela EI., Petrovic M., Gysling K., Pardo L., Franco R., Navarro G. Results from PLA experiments (Figs 5+6) have not been included due to ambiguity in the antibodies used. (anti-Ghrelin or anti -Ghrelin Receptor.) rfranco123@gmail.com,rfranco@ub.edu,dimartts@hotmail.com The email address for Gemma Navarro does not work; nor does g.navarro@ub.edu Molecular neurobiology 2019 Only protein-protein interactions imex curation imex curation aguinaga-2019-1 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 Aguinaga D., Medrano M., Cordomí A., Jiménez-Rosés M., Angelats E., Casanovas M., Vega-Quiroga I., Canela EI., Petrovic M., Gysling K., Pardo L., Franco R., Navarro G. Results from PLA experiments (Figs 5+6) have not been included due to ambiguity in the antibodies used. (anti-Ghrelin or anti -Ghrelin Receptor.) rfranco123@gmail.com,rfranco@ub.edu,dimartts@hotmail.com The email address for Gemma Navarro does not work; nor does g.navarro@ub.edu Molecular neurobiology 2019 Only protein-protein interactions imex curation imex curation aguinaga-2019-2 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 Aguinaga D., Medrano M., Cordomí A., Jiménez-Rosés M., Angelats E., Casanovas M., Vega-Quiroga I., Canela EI., Petrovic M., Gysling K., Pardo L., Franco R., Navarro G. Results from PLA experiments (Figs 5+6) have not been included due to ambiguity in the antibodies used. (anti-Ghrelin or anti -Ghrelin Receptor.) rfranco123@gmail.com,rfranco@ub.edu,dimartts@hotmail.com The email address for Gemma Navarro does not work; nor does g.navarro@ub.edu Molecular neurobiology 2019 Only protein-protein interactions imex curation imex curation aguinaga-2019-4 rat-striatal neurons Rattus norvegicus brain-striatum_neuron striatal neurons pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Aguinaga D., Medrano M., Cordomí A., Jiménez-Rosés M., Angelats E., Casanovas M., Vega-Quiroga I., Canela EI., Petrovic M., Gysling K., Pardo L., Franco R., Navarro G. Results from PLA experiments (Figs 5+6) have not been included due to ambiguity in the antibodies used. (anti-Ghrelin or anti -Ghrelin Receptor.) rfranco123@gmail.com,rfranco@ub.edu,dimartts@hotmail.com The email address for Gemma Navarro does not work; nor does g.navarro@ub.edu Molecular neurobiology 2019 Only protein-protein interactions imex curation imex curation q92847-1 Growth hormone secretagogue receptor type 1 GH-releasing peptide receptor Ghrelin receptor GHSR protein protein human Homo sapiens Human MWNATPSEEPGFNLTLADLDWDASPGNDSLGDELLQLFPAPLLAGVTATCVALFVVGIAGNLLTMLVVSRFRELRTTTNLYLSSMAFSDLLIFLCMPLDLVRLWQYRPWNFGDLLCKLFQFVSESCTYATVLTITALSVERYFAICFPLRAKVVVTKGRVKLVIFVIWAVAFCSAGPIFVLVGVEHENGTDPWDTNECRPTEFAVRSGLLTVMVWVSSIFFFLPVFCLTVLYSLIGRKLWRRRRGDAVVGASLRDQNHKQTVKMLAVVVFAFILCWLPFHVGRYLFSKSFEPGSLEIAQISQYCNLVSFVLFYLSAAINPILYNIMSKKYRVAVFRLLGFEPFSQRKLSTLKDESSRAWTESSINT q92847-2 Growth hormone secretagogue receptor type 1 GH-releasing peptide receptor Ghrelin receptor GHSR protein protein human Homo sapiens Human MWNATPSEEPGFNLTLADLDWDASPGNDSLGDELLQLFPAPLLAGVTATCVALFVVGIAGNLLTMLVVSRFRELRTTTNLYLSSMAFSDLLIFLCMPLDLVRLWQYRPWNFGDLLCKLFQFVSESCTYATVLTITALSVERYFAICFPLRAKVVVTKGRVKLVIFVIWAVAFCSAGPIFVLVGVEHENGTDPWDTNECRPTEFAVRSGLLTVMVWVSSIFFFLPVFCLTVLYSLIGRKLWRRRRGDAVVGASLRDQNHKQTVKMLGGSQRALRLSLAGPILSLCLLPSL sgmr1_human Sigma non-opioid intracellular receptor 1 SIGMAR1 OPRS1 SRBP Aging-associated gene 8 protein SR31747-binding protein Sigma 1-type opioid receptor AAG8 protein protein human Homo sapiens Human MQWAVGRRWAWAALLLAVAAVLTQVVWLWLGTQSFVFQREEIAQLARQYAGLDHELAFSRLIVELRRLHPGHVLPDEELQWVFVNAGGWMGAMCLLHASLSEYVLLFGTALGSRGHSGRYWAEISDTIISGTFHQWREGTTKSEVFYPGETVVHGPGEATAVEWGPNTWMVEYGRGVIPSTLAFALADTVFSTQDFLTLFYTLRSYARGLRLELTTYLFGQDP ghrl_rat Appetite-regulating hormone Growth hormone secretagogue Growth hormone-releasing peptide Motilin-related peptide Ghrl protein protein rat Rattus norvegicus (Rat) Rat MVSSATICSLLLLSMLWMDMAMAGSSFLSPEHQKAQQRKESKKPPAKLQPRALEGWLHPEDRGQAEEAEEELEIRFNAPFDVGIKLSGAQYQQHGRALGKFLQDILWEEVKEAPANK sgmr1_rat Sigma non-opioid intracellular receptor 1 Sigma 1-type opioid receptor Sigmar1 Oprs1 protein protein rat Rattus norvegicus (Rat) Rat MPWAVGRRWAWITLFLTIVAVLIQAVWLWLGTQSFVFQREEIAQLARQYAGLDHELAFSRLIVELRRLHPGHVLPDEELQWVFVNAGGWMGAMCLLHASLSEYVLLFGTALGSHGHSGRYWAEISDTIISGTFHQWREGTTKSEVYYPGETVVHGPGEATAVEWGPNTWMVEYGRGVIPSTLAFALSDTIFSTQDFLTLFYTLRAYARGLRLELTTYLFGQDP ghsr_rat Growth hormone secretagogue receptor type 1 GH-releasing peptide receptor Ghrelin receptor Ghsr protein protein rat Rattus norvegicus (Rat) Rat MWNATPSEEPEPNVTLDLDWDASPGNDSLPDELLPLFPAPLLAGVTATCVALFVVGISGNLLTMLVVSRFRELRTTTNLYLSSMAFSDLLIFLCMPLDLVRLWQYRPWNFGDLLCKLFQFVSESCTYATVLTITALSVERYFAICFPLRAKVVVTKGRVKLVILVIWAVAFCSAGPIFVLVGVEHENGTDPRDTNECRATEFAVRSGLLTVMVWVSSVFFFLPVFCLTVLYSLIGRKLWRRRGDAAVGASLRDQNHKQTVKMLAVVVFAFILCWLPFHVGRYLFSKSFEPGSLEIAQISQYCNLVSFVLFYLSAAINPILYNIMSKKYRVAVFKLLGFESFSQRKLSTLKDESSRAWTKSSINT ghsr-ghsr-1 3577220 N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2A-Control In the presence of interference peptides, fluorescence decreased by twofold only with TM5 and TM6 peptides. ghsr-ghsr-2 3577220 N/A 3577225 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2B/C-Control In the presence of interference peptides, fluorescence decreased by twofold only with TM5 and TM6 peptides. However in the presence of non-fused GHS-R1a, fluorescence was reduced in the presence of TM4, TM5, and TM6 peptides. ghsr-ghsr-3 3577220 N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2E-control In the presence of interference peptides, fluorescence decreased with TM1, TM2, and TM5 peptides. ghsr-sigmar1-3 3577220 N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577225 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2F/G control In the presence of interference peptides, fluorescence decreased only with TM1 and TM2 peptides. A similar result was obtained in the presence of unfused GHS-R1b. ghsr-ghsr-4 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM6_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2A-2 In the presence of interference peptides, fluorescence decreased by twofold only with TM5 and TM6 peptides. ghsr-ghsr-5 3577220 N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM5_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Figure 2A-1 In the presence of interference peptides, fluorescence decreased by twofold only with TM5 and TM6 peptides. ghsr-ghsr-6 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM5_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577225 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2B/C-1 In the presence of interference peptides, fluorescence decreased by twofold only with TM5 and TM6 peptides. However in the presence of non-fused GHS-R1a, fluorescence was reduced in the presence of TM4, TM5, and TM6 peptides. ghsr-ghsr-7 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM6_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577225 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2B/C-2 In the presence of interference peptides, fluorescence decreased by twofold only with TM5 and TM6 peptides. However in the presence of non-fused GHS-R1a, fluorescence was reduced in the presence of TM4, TM5, and TM6 peptides. ghsr-sigmar1-5 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM5_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2E-1 In the presence of interference peptides, fluorescence decreased with TM1, TM2, and TM5 peptides. ghsr-ghsr-8 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM1_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2E-2 In the presence of interference peptides, fluorescence decreased with TM1, TM2, and TM5 peptides. ghsr-ghsr-9 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM2_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2E-3 In the presence of interference peptides, fluorescence decreased with TM1, TM2, and TM5 peptides. ghsr-sigmar1-6 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM1_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577225 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2F/G-1 In the presence of interference peptides, fluorescence decreased only with TM1 and TM2 peptides. A similar result was obtained in the presence of unfused GHS-R1b. ghsr-ghsr-10 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM2_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577225 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2F/G-2 In the presence of interference peptides, fluorescence decreased only with TM1 and TM2 peptides. A similar result was obtained in the presence of unfused GHS-R1b. ghsr-ghsr-17 3577220 N/A 3577224 competitor competitor competitor unspecified role unspecified role purified purified TM4_peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3577225 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region cyfp cYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3577224 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed region nyfp nYFP undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Figure 2B/C-3 In the presence of interference peptides, fluorescence decreased by twofold only with TM5 and TM6 peptides. However in the presence of non-fused GHS-R1a, fluorescence was reduced in the presence of TM4, TM5, and TM6 peptides. ghsr-sigmar1-1 3577221 N/A 3577224 immunocytochemistry immunocytochemistry 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 false N/A 3577226 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed 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 colocalization colocalization false false false Figure 1A,3A cocaine PRE-084 -a Sigma 1R agonist Both Cocaine and PRE-084 increased colocalization of sigma- 1 R and GHS-R1a at the cell surface. ghsr-sigmar1-2 3577222 N/A 3577224 immunocytochemistry immunocytochemistry unspecified role unspecified role luminescence accept luminescence acceptor luminescence accept over-expressed over expressed level over-expressed 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 3577226 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 false proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 1C, 3B Addition of Cocaine or PRE-084 (Sigma-1R agonist) did not show any significant effect of treatments versus control. ghsr-sigmar1-4 3577223 N/A 3577227 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3577228 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3577229 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 false false false Figure 5C/D, 6 It is stated that "GHS-R1a-Sigma 1 R heteroreceptor complexes were identified in striatal primary cultures". The Methods section states use of "anti-GHS-R1a antibody (1/100, AbCam,Cambridge, UK)";however the image legend text states that "a polyclonal anti-ghrelin primary antibody (1/100, AbCam," was used...). The only Uniprot entry for Rat GHSR has been used here as that would appear to be the most probable protein detected. Cocaine Interaction was seen in thick striatal sections and also striatal primary cultures (neurons).