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/ The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. Biochimica et biophysica acta 2015 imex curation Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. pere.garriga@upc.edu 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient Human 293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient ATCC® CRL-3022 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. pere.garriga@upc.edu Accepted 2019-JUN-19 AT 13:22 BST AT 13:22 BST by ORCHARD Biochimica et biophysica acta 2015 Only protein-protein interactions imex curation imex curation The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. Biochimica et biophysica acta 2015 imex curation Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. pere.garriga@upc.edu 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient Human 293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient ATCC® CRL-3022 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. Details of CFP tagged 5HT1A are stated to be found in ref 15; only this publication does not mention "CFP". pere.garriga@upc.edu Accepted 2019-JUN-19 AT 13:22 BST AT 13:22 BST by ORCHARD Biochimica et biophysica acta 2015 Only protein-protein interactions imex curation imex curation The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. Biochimica et biophysica acta 2015 imex curation Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. pere.garriga@upc.edu 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient Human 293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient ATCC® CRL-3022 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. pere.garriga@upc.edu Accepted 2019-JUN-19 AT 13:22 BST AT 13:22 BST by ORCHARD Biochimica et biophysica acta 2015 Only protein-protein interactions imex curation imex curation The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. The zinc binding receptor GPR39 interacts with 5-HT1A and GalR1 to form dynamic heteroreceptor complexes with signaling diversity. Biochimica et biophysica acta 2015 imex curation Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. pere.garriga@upc.edu 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient Human 293s_gnti HEK293S GnTI -adenovirus 5 transformed, glycosylation deficient ATCC® CRL-3022 fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Tena-Campos M., Ramon E., Borroto-Escuela DO., Fuxe K., Garriga P. pere.garriga@upc.edu Accepted 2019-JUN-19 AT 13:22 BST AT 13:22 BST by ORCHARD Biochimica et biophysica acta 2015 Only protein-protein interactions imex curation imex curation 5ht1a_human 5-hydroxytryptamine receptor 1A HTR1A ADRB2RL1 ADRBRL1 G-21 Serotonin receptor 1A protein protein human Homo sapiens Human MDVLSPGQGNNTTSPPAPFETGGNTTGISDVTVSYQVITSLLLGTLIFCAVLGNACVVAAIALERSLQNVANYLIGSLAVTDLMVSVLVLPMAALYQVLNKWTLGQVTCDLFIALDVLCCTSSILHLCAIALDRYWAITDPIDYVNKRTPRRAAALISLTWLIGFLISIPPMLGWRTPEDRSDPDACTISKDHGYTIYSTFGAFYIPLLLMLVLYGRIFRAARFRIRKTVKKVEKTGADTRHGASPAPQPKKSVNGESGSRNWRLGVESKAGGALCANGAVRQGDDGAALEVIEVHRVGNSKEHLPLPSEAGPTPCAPASFERKNERNAEAKRKMALARERKTVKTLGIIMGTFILCWLPFFIVALVLPFCESSCHMPTLLGAIINWLGYSNSLLNPVIYAYFNKDFQNAFKKIIKCKFCRQ 762664FCF62CFD8F gpr39_human G-protein coupled receptor 39 GPR39 protein protein human Homo sapiens Human MASPSLPGSDCSQIIDHSHVPEFEVATWIKITLILVYLIIFVMGLLGNSATIRVTQVLQKKGYLQKEVTDHMVSLACSDILVFLIGMPMEFYSIIWNPLTTSSYTLSCKLHTFLFEACSYATLLHVLTLSFERYIAICHPFRYKAVSGPCQVKLLIGFVWVTSALVALPLLFAMGTEYPLVNVPSHRGLTCNRSSTRHHEQPETSNMSICTNLSSRWTVFQSSIFGAFVVYLVVLLSVAFMCWNMMQVLMKSQKGSLAGGTRPPQLRKSESEESRTARRQTIIFLRLIVVTLAVCWMPNQIRRIMAAAKPKHDWTRSYFRAYMILLPFSETFFYLSSVINPLLYTVSSQQFRRVFVQVLCCRLSLQHANHEKRLRVHAHSTTDSARFVQRPLLFASRRQSSARRTEKIFLSTFQSEAEPQSKSQSLSLESLEPNSGAKPANSAAENGFQEHEV mint homomint 8E3A233420D9021E galr1_human Galanin receptor type 1 GALR1 GALNR GALNR1 protein protein human Homo sapiens Human MELAVGNLSEGNASWPEPPAPEPGPLFGIGVENFVTLVVFGLIFALGVLGNSLVITVLARSKPGKPRSTTNLFILNLSIADLAYLLFCIPFQATVYALPTWVLGAFICKFIHYFFTVSMLVSIFTLAAMSVDRYVAIVHSRRSSSLRVSRNALLGVGCIWALSIAMASPVAYHQGLFHPRASNQTFCWEQWPDPRHKKAYVVCTFVFGYLLPLLLICFCYAKVLNHLHKKLKNMSKKSEASKKKTAQTVLVVVVVFGISWLPHHIIHLWAEFGVFPLTPASFLFRITAHCLAYSNSSVNPIIYAFLSENFRKAYKQVFKCHIRKDSHLSDTKESKSRIDTPPSTNCTHV A0664227157B3410 gpr39-htr1a-1 4 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed c-terminus eyfp enhanced yellow fluorescent protein tag eyfp 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 rho tag rho tag 1D4 tag rho1D4 tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 1 A-F Both receptors share a similar distributional pattern and significantly co-localized in the membrane cell surface and reticular compartments. gpr39-htr1a-2 2 6 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed c-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role fluorescence donor fluorescence donor 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 1G, 2B Both receptors share a similar distributional pattern and significantly co-localized in the membrane cell surface and reticular compartments. Using a saturation assay we assessed specific FRET between the two receptors, in cells co-expressing a constant amount of hrGFP2-5-HT1A (humanized green fluorescent protein 2 fused to the C-terminal tail of the 5-HT1A) (donor) and an increasing concentration of GPR39-EYFP (GPR39 receptor fused to enhanced yellow fluorescent protein) (acceptor). gpr39-htr1a-3 3 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed c-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminus rho tag rho tag 1D4 tag rho1D4 tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 2A-c htr1a-gpr39-1 1 6 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed c-terminus eyfp enhanced yellow fluorescent protein tag eyfp c-term range C-terminal range c-term range c-term range C-terminal range c-term range 7 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminus rho tag rho tag 1D4 tag rho1D4 tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range association association Figure 2A-d htr1a-gpr39-3 2 5 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed c-terminus eyfp enhanced yellow fluorescent protein tag eyfp 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 2B 5-HT1A maintained its capacity to interact with GalR1in the presence of GPR39.