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/ Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Mol. Cell. Biol. (0270-7306) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. davet Only protein-protein interactions 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 Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 davet Only protein-protein interactions imex curation Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Mol. Cell. Biol. (0270-7306) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. davet Only protein-protein interactions 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 Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 davet Only protein-protein interactions imex curation Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Mol. Cell. Biol. (0270-7306) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. davet Only protein-protein interactions 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 Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 davet Only protein-protein interactions imex curation Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Mol. Cell. Biol. (0270-7306) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. davet 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 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 Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 davet Only protein-protein interactions imex curation Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Mol. Cell. Biol. (0270-7306) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. davet Only protein-protein interactions 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 Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 davet Only protein-protein interactions imex curation Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Mol. Cell. Biol. (0270-7306) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. davet Only protein-protein interactions 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 Accepted 2007-Not a month!-19 by DAVET. A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 davet Only protein-protein interactions imex curation Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Identification and characterization of putative tumor suppressor NGB, a GTP-binding protein that interacts with the neurofibromatosis 2 protein. Mol. Cell. Biol. (0270-7306) 2007 imex curation 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 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. davet Only protein-protein interactions 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 Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. A human brain cDNA library was used. Mol. Cell. Biol. (0270-7306) 2007 davet Only protein-protein interactions imex curation 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 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 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 0A05E65E65D6DB62 merl_human Merlin Moesin-ezrin-radixin-like protein Neurofibromin-2 Schwannomin Schwannomerlin NF2 SCH protein protein human Homo sapiens Human MAGAIASRMSFSSLKRKQPKTFTVRIVTMDAEMEFNCEMKWKGKDLFDLVCRTLGLRETWFFGLQYTIKDTVAWLKMDKKVLDHDVSKEEPVTFHFLAKFYPENAEEELVQEITQHLFFLQVKKQILDEKIYCPPEASVLLASYAVQAKYGDYDPSVHKRGFLAQEELLPKRVINLYQMTPEMWEERITAWYAEHRGRARDEAEMEYLKIAQDLEMYGVNYFAIRNKKGTELLLGVDALGLHIYDPENRLTPKISFPWNEIRNISYSDKEFTIKPLDKKIDVFKFNSSKLRVNKLILQLCIGNHDLFMRRRKADSLEVQQMKAQAREEKARKQMERQRLAREKQMREEAERTRDELERRLLQMKEEATMANEALMRSEETADLLAEKAQITEEEAKLLAQKAAEAEQEMQRIKATAIRTEEEKRLMEQKVLEAEVLALKMAEESERRAKEADQLKQDLQEAREAERRAKQKLLEIATKPTYPPMNPIPAPLPPDIPSFNLIGDSLSFDFKDTDMKRLSMEIEKEKVEYMEKSKHLQEQLNELKTEIEALKLKERETALDILHNENSDRGGSSKHNTIKKLTLQSAKSRVAFFEEL B1A1BF2BD5DA561C guanosine 5'-[gamma-thio]triphosphate GTP-gamma-S small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) nf2-ngb 6 9 unspecified role unspecified role prey prey 10 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 physical association physical association A human brain cDNA library was used in this screen because merlin is highly expressed in the brain (44, 58). An N-terminal portion (amino acids 1 to 374) of merlin was cloned into the EcoRI and BamHI sites of pJK202 to create the bait pNLexA-NF2N. Three clones that specifically interacted with the bait were identified. Sequence analysis revealed that two of the clones contained overlapping sequences of a cDNA (26). The largest clone contained a 243-amino-acid open reading frame with a conserved GTP-binding domain named NGB (NF2-associated GTP binding protein) 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 4 9 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 10 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 physical association physical association BAIT gene AF120334 P35240; Q9NPG2 transfection HA-tagged NF2 and Flag-tagged NGB were cotransfected into HEK293 cells. After 48 h of the transfection, the cells were lysed and immunoprecipitated with anti-HA (NF2) or anti-Flag (NGB) antibody. The immunoprecipitates were subjected to Western blot analysis. As shown in Fig. 2A and B, Flag-NGB and HA-NF2 were readily detected in anti-HA and anti-Flag immunoprecipitates, respectively. 2 A nf2-ngb-1 4 9 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 10 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 physical association physical association PREY gene AF120334 P35240; Q9NPG2 transfection HA-tagged NF2 and Flag-tagged NGB were cotransfected into HEK293 cells. After 48 h of the transfection, the cells were lysed and immunoprecipitated with anti-HA (NF2) or anti-Flag (NGB) antibody. The immunoprecipitates were subjected to Western blot analysis. As shown in Fig. 2A and B, Flag-NGB and HA-NF2 were readily detected in anti-HA and anti-Flag immunoprecipitates, respectively. 2 B nf2-ngb-2 7 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association PREY gene AF120334 2 E To demonstrate that endogenous NGB and NF2 interact, a coimmunoprecipitation experiment was performed using both 82HTB cells and fresh skeletal muscle tissues with antibodies against NF2 and NGB. The specificity of anti-NGB antibody was demonstrated by Western and immunoprecipitation (IP)-Western blot analysis (Fig. 2C and D). Figure 2E shows that NGB was immunoprecipitated with anti-NF2 antibody but not with preimmune serum. The immunoprecipitation of NGB was competed by GST-NF2 fusion protein. ngb-nf2-1 7 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association PREY gene AF120334 2 F To demonstrate that endogenous NGB and NF2 interact, a coimmunoprecipitation experiment was performed using both 82HTB cells and fresh skeletal muscle tissues with antibodies against NF2 and NGB. The specificity of anti-NGB antibody was demonstrated by Western and immunoprecipitation (IP)-Western blot analysis (Fig. 2C and D). Figure 2E shows that NGB was immunoprecipitated with anti-NF2 antibody but not with preimmune serum. Moreover, the immunoprecipitation of NGB was competed by GST-NF2 fusion protein, which was used as an antigen to generate the anti-NF2 antibody. Further, merlin was detected in NGB immunoprecipitates The immunoprecipitation of NGB was competed by GST-NF2 fusion protein. ngb-nf2-2 5 9 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component physical association physical association Q9NPG2 gene AF120334 2 H NGB and merlin predominantly colocalized in the perinuclear cytoplasm ngb-nf2-3 2 9 unspecified role unspecified role prey prey 10 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 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association PREY gene AF120334 2 I Yeast two-hybrid tests of interaction revealed that the amino (aa 1 to 52) and carboxyl (aa 288 to 344) termini of merlin are required for binding to NGB (Fig. 2I). 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 2 9 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 10 unspecified role unspecified role bait bait physical association physical association PREY gene AF120334 2 J The binding region of NGB was mapped to the G-protein homology domain ngb-nf2-5 4 9 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 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 R A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:53:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg394ala,lys395ala' Tue Jun 30 15:36:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg394Ala;Lys395Ala]' 10 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 physical association physical association BAIT gene AF120334 P35240; Q9NPG2 transfection 2 K Immunoprecipitation experiments showed that mutation of lysine-395/arginine-394 to alanine (NGB-K395/R394A) significantly reduced the ability of NGB to bind to merlin ngb-gtp_gamma_s 1 9 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 chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 11 unspecified role unspecified role neutral component neutral component physical association physical association Q9NPG2 gene AF120334 7 A, D NGB binds to GTPS within 2 min. 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 3 9 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 chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 8 enzyme target enzyme target substrate substrate unspecified role unspecified role physical association physical association ENZYME gene AF120334 7 B, C The GTP was [alpha32P]-labelled. Repeated experiments revealed that NGB was capable of hydrolyzing [-32P]GTP in the presence of an excess of unlabeled UTP but not GTP An excess of unlabelled GTP inhibited this interaction. This interaction also occurred in the presence of Merlin.