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/ lee-2007a-2 Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. 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 anti tag western anti tag western blot anti tag western Lee H., Kim D., Dan HC., Wu EL., Gritsko TM., Cao C., Nicosia SV., Golemis EA., Liu W., Coppola D., Brem SS., Testa JR., Cheng JQ. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with ubiquitin since the organism of origin of this interactor cannot be ascertained. Thus interactions involving ubiquitin could not be added. Joseph.Testa@fccc.edu,ChengJQ@moffitt.usf.edu A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 Only protein-protein interactions imex curation lee-2007a-3 Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. human-a_204 Homo sapiens rhabdomyosarcoma cell line A-204 a_204 Human rhabdomyosarcoma cell line A-204 Established from the rhabdomyosarcoma of a 1-year-old girl in 1973; reported to form small malignant tumors in nude mice consistent with embryonic rhabdomyosarcoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lee H., Kim D., Dan HC., Wu EL., Gritsko TM., Cao C., Nicosia SV., Golemis EA., Liu W., Coppola D., Brem SS., Testa JR., Cheng JQ. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with ubiquitin since the organism of origin of this interactor cannot be ascertained. Thus interactions involving ubiquitin could not be added. Joseph.Testa@fccc.edu,ChengJQ@moffitt.usf.edu A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 Only protein-protein interactions imex curation lee-2007a-5 Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Lee H., Kim D., Dan HC., Wu EL., Gritsko TM., Cao C., Nicosia SV., Golemis EA., Liu W., Coppola D., Brem SS., Testa JR., Cheng JQ. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with ubiquitin since the organism of origin of this interactor cannot be ascertained. Thus interactions involving ubiquitin could not be added. Joseph.Testa@fccc.edu,ChengJQ@moffitt.usf.edu A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 Only protein-protein interactions imex curation lee-2007a-1 Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Lee H., Kim D., Dan HC., Wu EL., Gritsko TM., Cao C., Nicosia SV., Golemis EA., Liu W., Coppola D., Brem SS., Testa JR., Cheng JQ. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with ubiquitin since the organism of origin of this interactor cannot be ascertained. Thus interactions involving ubiquitin could not be added. Joseph.Testa@fccc.edu,ChengJQ@moffitt.usf.edu A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 Only protein-protein interactions imex curation lee-2007a-4 Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. human-a_204 Homo sapiens rhabdomyosarcoma cell line A-204 a_204 Human rhabdomyosarcoma cell line A-204 Established from the rhabdomyosarcoma of a 1-year-old girl in 1973; reported to form small malignant tumors in nude mice consistent with embryonic rhabdomyosarcoma confocal microscopy confocal microscopy predetermined predetermined participant predetermined Lee H., Kim D., Dan HC., Wu EL., Gritsko TM., Cao C., Nicosia SV., Golemis EA., Liu W., Coppola D., Brem SS., Testa JR., Cheng JQ. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with ubiquitin since the organism of origin of this interactor cannot be ascertained. Thus interactions involving ubiquitin could not be added. Joseph.Testa@fccc.edu,ChengJQ@moffitt.usf.edu A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 Only protein-protein interactions imex curation lee-2007a-6 Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay predetermined predetermined participant predetermined Lee H., Kim D., Dan HC., Wu EL., Gritsko TM., Cao C., Nicosia SV., Golemis EA., Liu W., Coppola D., Brem SS., Testa JR., Cheng JQ. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with ubiquitin since the organism of origin of this interactor cannot be ascertained. Thus interactions involving ubiquitin could not be added. Joseph.Testa@fccc.edu,ChengJQ@moffitt.usf.edu A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 Only protein-protein interactions imex curation lee-2007a-7 Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. in vitro In vitro chromatography chromatography technology column chromatography chromatography predetermined predetermined participant predetermined Lee H., Kim D., Dan HC., Wu EL., Gritsko TM., Cao C., Nicosia SV., Golemis EA., Liu W., Coppola D., Brem SS., Testa JR., Cheng JQ. Cancer - Interactions investigated in the context of cancer NGB was incubated with [alpha-32P]GTP (10 microM; 2 Ci/mmol). Aliquots were removed from the reaction mixture at the indicated times, analyzed by poly(ethyleneimine)thin-layer chromatography plates, and exposed to film for 5 min. A unique UniProtKB entry could not be associated with ubiquitin since the organism of origin of this interactor cannot be ascertained. Thus interactions involving ubiquitin could not be added. Joseph.Testa@fccc.edu,ChengJQ@moffitt.usf.edu A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 Only protein-protein interactions imex curation gtpb4_human GTP-binding protein 4 GTPBP4 CRFG NOG1 Chronic renal failure gene protein GTP-binding protein NGB Nucleolar GTP-binding protein 1 protein protein human Homo sapiens Human MAHYNFKKITVVPSAKDFIDLTLSKTQRKTPTVIHKHYQIHRIRHFYMRKVKFTQQNYHDRLSQILTDFPKLDDIHPFYADLMNILYDKDHYKLALGQINIAKNLVDNVAKDYVRLMKYGDSLYRCKQLKRAALGRMCTVIKRQKQSLEYLEQVRQHLSRLPTIDPNTRTLLLCGYPNVGKSSFINKVTRADVDVQPYAFTTKSLFVGHMDYKYLRWQVVDTPGILDHPLEDRNTIEMQAITALAHLRAAVLYVMDLSEQCGHGLREQLELFQNIRPLFINKPLIVVANKCDVKRIAELSEDDQKIFTDLQSEGFPVIETSTLTEEGVIKVKTEACDRLLAHRVETKMKGNKVNEVLNRLHLAIPTRRDDKERPPFIPEGVVARRKRMETEESRKKRERDLELEMGDDYILDLQKYWDLMNLSEKHDKIPEIWEGHNIADYIDPAIMKKLEELEKEEELRTAAGEYDSVSESEDEEMLEIRQLAKQIREKKKLKILESKEKNTQGPRMPRTAKKVQRTVLEKEMRSLGVDMDDKDDAHYAVQARRSRSITRKRKREDSAPPSSVARSGSCSRTPRDVSGLRDVKMVKKAKTMMKNAQKKMNRLGKKGEADRHVFDMKPKHLLSGKRKAGKKDRR merl_human Merlin Moesin-ezrin-radixin-like protein Neurofibromin-2 Schwannomin Schwannomerlin NF2 SCH protein protein human Homo sapiens Human MAGAIASRMSFSSLKRKQPKTFTVRIVTMDAEMEFNCEMKWKGKDLFDLVCRTLGLRETWFFGLQYTIKDTVAWLKMDKKVLDHDVSKEEPVTFHFLAKFYPENAEEELVQEITQHLFFLQVKKQILDEKIYCPPEASVLLASYAVQAKYGDYDPSVHKRGFLAQEELLPKRVINLYQMTPEMWEERITAWYAEHRGRARDEAEMEYLKIAQDLEMYGVNYFAIRNKKGTELLLGVDALGLHIYDPENRLTPKISFPWNEIRNISYSDKEFTIKPLDKKIDVFKFNSSKLRVNKLILQLCIGNHDLFMRRRKADSLEVQQMKAQAREEKARKQMERQRLAREKQMREEAERTRDELERRLLQMKEEATMANEALMRSEETADLLAEKAQITEEEAKLLAQKAAEAEQEMQRIKATAIRTEEEKRLMEQKVLEAEVLALKMAEESERRAKEADQLKQDLQEAREAERRAKQKLLEIATKPTYPPMNPIPAPLPPDIPSFNLIGDSLSFDFKDTDMKRLSMEIEKEKVEYMEKSKHLQEQLNELKTEIEALKLKERETALDILHNENSDRGGSSKHNTIKKLTLQSAKSRVAFFEEL guanosine 5'-[gamma-thio]triphosphate GTP-gamma-S small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 ngb-nf2 FLAG-NGB coimmunoprecipitates HA-Merlin in HEK293 lysates 663122 N/A 663129 unspecified role unspecified role bait bait region 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 false N/A 663130 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false BAIT gene AF120334 P35240; Q9NPG2 transfection 2 A nf2-ngb-1 FLAG-NGB coimmunoprecipitates with HA-Merlin in HEK293 lysates 663122 N/A 663129 unspecified role unspecified role prey prey region 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 false N/A 663130 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false PREY gene AF120334 P35240; Q9NPG2 transfection 2 B ngb-nf2-5 GFP-NGB coimmunoprecipitates HA-Merlin in HEK293 lysates 663122 N/A 663129 unspecified role unspecified role bait bait 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 Q9BZE4:p.[Arg394Ala;Lys395Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 663130 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false BAIT gene AF120334 P35240; Q9NPG2 transfection 2 K nf2-ngb-2 NGB coimmunoprecipitates with Merlin in 82HTB (A-204) cell lysates 663123 N/A 663129 unspecified role unspecified role prey prey N/A 663130 unspecified role unspecified role bait bait physical association physical association false false false PREY gene AF120334 2 E The immunoprecipitation of NGB was competed by GST-NF2 fusion protein. ngb-nf2-1 NGB coimmunoprecipitates Merlin in 82HTB (A-204) cell lysates 663123 N/A 663129 unspecified role unspecified role bait bait N/A 663130 unspecified role unspecified role prey prey physical association physical association false false false PREY gene AF120334 2 F The immunoprecipitation of NGB was competed by GST-NF2 fusion protein. ngb-nf2-3 NGB interacts with Merlin in a yeast two-hybrid assay 663124 N/A 663129 unspecified role unspecified role prey prey N/A 663130 unspecified role unspecified role bait bait n-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false PREY gene AF120334 2 I Neither the amino terminus nor the carboxyl region alone binds to NGB, suggesting that interdomain association between the amino and carboxyl termini of merlin, which is essential for its tumor suppressor activity. ngb-nf2-4 NGB interacts with Merlin in a yeast two-hybrid assay 663124 N/A 663129 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 663130 unspecified role unspecified role bait bait physical association physical association false false false PREY gene AF120334 2 J nf2-ngb 663125 N/A 663129 unspecified role unspecified role prey prey N/A 663130 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false PREY gene AF120334 The largest clone is stated to contain a 243 aa open reading frame with a conserved GTP-binding domain (aa 171-350). ngb-nf2-2 NGB and Merlin colocalize in 82HTB (A-204) cells 663126 N/A 663129 unspecified role unspecified role neutral component neutral component N/A 663130 unspecified role unspecified role neutral component neutral component physical association physical association false false false Q9NPG2 gene AF120334 2 H ngb-gtp_gamma_s NGB binds GTPgammaS 663127 N/A 663129 unspecified role unspecified role neutral component neutral component region 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 false chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 N/A 663131 unspecified role unspecified role neutral component neutral component physical association physical association false false false Q9NPG2 gene AF120334 7 A, D The GTPgammaS was [35S]-labelled. The NGB-[35S]GTPgammaS binding activity was completely blocked by an excess (20-fold) of unlabeled GTP or GDP but not ATP, UTP, or CTP, indicating that NGB is a specific guanine nucleotide-binding protein. The NGB-[35S]GTPgammaS binding activity was completely blocked by an excess (20-fold) of unlabeled GTP or GDP. THe GTP-binding activity of NGB is not affected by Merlin. ngb-gtp NGB has intrinsic GTPase activity 663128 N/A 663129 enzyme enzyme unspecified role unspecified role region 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 false chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 N/A 663132 enzyme target enzyme target substrate substrate unspecified role unspecified role physical association physical association false false false ENZYME gene AF120334 7 B, C The GTP was [alpha32P]-labelled. An excess of unlabelled GTP inhibited this interaction. This interaction also occurred in the presence of Merlin.