MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it The orphan GPR50 receptor specifically inhibits MT1 melatonin receptor function through heterodimerization. The orphan GPR50 receptor specifically inhibits MT1 melatonin receptor function through heterodimerization. EMBO J. (0261-4189) 2006 imex curation Levoye A., Dam J., Ayoub MA., Guillaume JL., Couturier C., Delagrange P., Jockers R. 2009-01-08 Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Levoye A., Dam J., Ayoub MA., Guillaume JL., Couturier C., Delagrange P., Jockers R. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation The orphan GPR50 receptor specifically inhibits MT1 melatonin receptor function through heterodimerization. The orphan GPR50 receptor specifically inhibits MT1 melatonin receptor function through heterodimerization. EMBO J. (0261-4189) 2006 imex curation Levoye A., Dam J., Ayoub MA., Guillaume JL., Couturier C., Delagrange P., Jockers R. 2009-01-08 Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Levoye A., Dam J., Ayoub MA., Guillaume JL., Couturier C., Delagrange P., Jockers R. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation The orphan GPR50 receptor specifically inhibits MT1 melatonin receptor function through heterodimerization. The orphan GPR50 receptor specifically inhibits MT1 melatonin receptor function through heterodimerization. EMBO J. (0261-4189) 2006 imex curation Levoye A., Dam J., Ayoub MA., Guillaume JL., Couturier C., Delagrange P., Jockers R. 2009-01-08 Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Levoye A., Dam J., Ayoub MA., Guillaume JL., Couturier C., Delagrange P., Jockers R. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation mt2_human Metallothionein-2 Metallothionein-II Metallothionein-2A MT2A CES1 MT2 protein protein human Homo sapiens Human MDPNCSCAAGDSCTCAGSCKCKECKCTSCKKSCCSCCPVGCAKCAQGCICKGASDKCSCCA 7B0A7E1F99843078 mtr1l_human Melatonin-related receptor GPR50 G protein-coupled receptor 50 H9 protein protein human Homo sapiens Human MGPTLAVPTPYGCIGCKLPQPEYPPALIIFMFCAMVITIVVDLIGNSMVILAVTKNKKLRNSGNIFVVSLSVADMLVAIYPYPLMLHAMSIGGWDLSQLQCQMVGFITGLSVVGSIFNIVAIAINRYCYICHSLQYERIFSVRNTCIYLVITWIMTVLAVLPNMYIGTIEYDPRTYTCIFNYLNNPVFTVTIVCIHFVLPLLIVGFCYVRIWTKVLAARDPAGQNPDNQLAEVRNFLTMFVIFLLFAVCWCPINVLTVLVAVSPKEMAGKIPNWLYLAAYFIAYFNSCLNAVIYGLLNENFRREYWTIFHAMRHPIIFFSGLISDIREMQEARTLARARAHARDQAREQDRAHACPAVEETPMNVRNVPLPGDAAAGHPDRASGHPKPHSRSSSAYRKSASTHHKSVFSHSKAASGHLKPVSGHSKPASGHPKSATVYPKPASVHFKADSVHFKGDSVHFKPDSVHFKPASSNPKPITGHHVSAGSHSKSAFSAATSHPKPTTGHIKPATSHAEPTTADYPKPATTSHPKPTAADNPELSASHCPEIPAIAHPVSDDSDLPESASSPAAGPTKPAASQLESDTIADLPDPTVVTTSTNDYHDVVVIDVEDDPDEMAV mint Does not bind melatonin A0B73A7ACE1053BB gnai1_human Guanine nucleotide-binding protein G(i) subunit alpha-1 GNAI1 Adenylate cyclase-inhibiting G alpha protein protein protein human Homo sapiens Human MGCTLSAEDKAAVERSKMIDRNLREDGEKAAREVKLLLLGAGESGKSTIVKQMKIIHEAGYSEEECKQYKAVVYSNTIQSIIAIIRAMGRLKIDFGDSARADDARQLFVLAGAAEEGFMTAELAGVIKRLWKDSGVQACFNRSREYQLNDSAAYYLNDLDRIAQPNYIPTQQDVLRTRVKTTGIVETHFTFKDLHFKMFDVGGQRSERKKWIHCFEGVTAIIFCVALSDYDLVLAEDEEMNRMHESMKLFDSICNNKWFTDTSIILFLNKKDLFEEKIKKSPLTICYPEYAGSNTYEEAAAYIQCQFEDLNKRKDTKEIYTHFTCATDTKNVQFVFDAVTDVIIKNNLKDCGLF 9F88311B46E62DE3 arrb1_human Beta-arrestin-1 Arrestin beta-1 ARRB1 ARR1 Non-visual arrestin-2 protein protein human Homo sapiens Human MGDKGTRVFKKASPNGKLTVYLGKRDFVDHIDLVDPVDGVVLVDPEYLKERRVYVTLTCAFRYGREDLDVLGLTFRKDLFVANVQSFPPAPEDKKPLTRLQERLIKKLGEHAYPFTFEIPPNLPCSVTLQPGPEDTGKACGVDYEVKAFCAENLEEKIHKRNSVRLVIRKVQYAPERPGPQPTAETTRQFLMSDKPLHLEASLDKEIYYHGEPISVNVHVTNNTNKTVKKIKISVRQYADICLFNTAQYKCPVAMEEADDTVAPSSTFCKVYTLTPFLANNREKRGLALDGKLKHEDTNLASSTLLREGANREILGIIVSYKVKVKLVVSRGGLLGDLASSDVAVELPFTLMHPKPKEEPPHREVPENETPVDTNLIELDTNDDDIVFEDFARQRLKGMKDDKEEEEDGTGSPQLNNR DIP protein P49407 original sequence version: 138 DIP protein P49407 original sequence version: 69 DIP protein P49407 original sequence version: 92 DIP protein P49407 original sequence version: 103 0A3C135092338D10 mt1a_human Metallothionein-1A MT1A MT1S Metallothionein-IA protein protein human Homo sapiens Human MDPNCSCATGGSCTCTGSCKCKECKCTSCKKSCCSCCPMSCAKCAQGCICKGASEKCSCCA mint Metallothioneins have a high content of cysteine residues that bind various heavy metals; these proteins are transcriptionally regulated by both heavy metals and glucocorticoids 8FBA7C54EE8B6A13 gpr50-gpr50 3 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tagged flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association F1B EMBO-Journal To confirm GPR50 homodimerization, we performed co-immunoprecipitation experiments with differentially tagged receptors. When immunoprecipitating GPR50-YFP, co-precipitated Flag-GPR50 was revealed in SDS PAGE as a monomer at 70 kDa and as a SDS-resistant dimer at 140 kDa (Figure 1B). homomint mint gpr50-mt1a 3 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tagged flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 physical association physical association F1C EMBO-Journal To explore GPR50 heterodimerization with MT1 and MT2, GPR50-YFP was coexpressed with the N-terminally Flag-tagged MT1 and Myc-tagged MT2. Anti-GFP antibodies were indeed able to specifically pull down MT1 and MT2 proteins (Figure 1C and D). homomint mint gpr50-mt2a 3 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 physical association physical association F1D EMBO-Journal To explore GPR50 heterodimerization with MT1 and MT2, GPR50-YFP was coexpressed with the N-terminally Flag-tagged MT1 and Myc-tagged MT2. Anti-GFP antibodies were indeed able to specifically pull down MT1 and MT2 proteins (Figure 1C and D). homomint mint gpr50-gnai1 3 6 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 15:05:50 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation decreasing' 1-349 range MINT-2836383: original sequence: 1-349 : MGPTLAVPTPYGCIGCKLPQPEYPPALIIFMFCAMVITIVVDLIGNSMVILAVTKNKKLRNSGNIFVVSLSVADMLVAIYPYPLMLHAMSIGGWDLSQLQCQMVGFITGLSVVGSIFNIVAIAINRYCYICHSLQYERIFSVRNTCIYLVITWIMTVLAVLPNMYIGTIEYDPRTYTCIFNYLNNPVFTVTIVCIHFVLPLLIVGFCYVRIWTKVLAARDPAGQNPDNQLAEVRNFLTMFVIFLLFAVCWCPINVLTVLVAVSPKEMAGKIPNWLYLAAYFIAYFNSCLNAVIYGLLNENFRREYWTIFHAMRHPIIFFPGLISDIREMQEARTLARARAHARDQARE yfp tagged 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 Stoichiometry: 1.0 physical association physical association F9A EMBO-Journal Gialpha was also observed in co-immunoprecipitates from GPR50-YFP- and GPR50DeltaCter-YFP-containing cell lysates (Figure 9A, lanes 2 and 3). At comparable receptor levels, the amount of Gialpha associated with GPR50-YFP was lower compared to MT1-YFP lysates and even lower for GPR50DeltaCter-YFP. homomint mint mt1a-gnai1 3 6 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 Stoichiometry: 1.0 physical association physical association F9A EMBO-Journal Cell lysates containing equivalent quantities of YFP fusion proteins were immunoprecipitated and the presence of Gialpha was revealed by Western blotting. Gialpha was readily co-precipitated from melatonin-stimulated MT1-YFP-containing cell lysates (Figure 9A, lane 1). homomint mint gpr50-gpr50-1 2 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 proximity proximity close-contact colocalization neighbourhood interaction F2 EMBO-Journal The GPR50-Rluc construct was used as BRET donor and was coexpressed with the BRET acceptor YFP fused to the C-terminus of different receptors (Figure 2A). Significant energy transfer was observed in cells coexpressing GPR50-Rluc with similar amounts of GPR50-YFP, MT1-YFP or MT2-YFP. homomint mint gpr50-mt2a-1 2 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction F2 EMBO-Journal The GPR50-Rluc construct was used as BRET donor and was coexpressed with the BRET acceptor YFP fused to the C-terminus of different receptors (Figure 2A). Significant energy transfer was observed in cells coexpressing GPR50-Rluc with similar amounts of GPR50-YFP, MT1-YFP or MT2-YFP. homomint mint gpr50-mt1a-1 2 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction F2 EMBO-Journal The GPR50-Rluc construct was used as BRET donor and was coexpressed with the BRET acceptor YFP fused to the C-terminus of different receptors (Figure 2A). Significant energy transfer was observed in cells coexpressing GPR50-Rluc with similar amounts of GPR50-YFP, MT1-YFP or MT2-YFP. homomint mint arrb1-mt1a 2 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 Stoichiometry: 1.0 proximity proximity close-contact colocalization neighbourhood interaction F9B F9C EMBO-Journal Dynamic recruitment of beta-arrestin 1 to MT1 was observed upon melatonin stimulation. Coexpression of Flag-GPR50 decreased the melatonin-induced BRET signal Dynamic recruitment of beta-arrestin 1 to MT1 was observed upon melatonin stimulation of cells coexpressing MT1-Rluc and an N-terminally YFP-tagged beta-arrestin 1 fusion protein (Figure 9B). homomint mint gpr50-mt1a-2 1 8 immunostaining immunostaining predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell flag tagged flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed living cell living cell yfp tagged 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 colocalization colocalization F7E EMBO-Journal GPR50-YFP itself localized to the cell membrane when expressed alone or with Flag-MT1 (Figure 7B, D and G). homomint mint