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 yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The Journal of cell biology 2007 imex curation Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk Accepted 2017-MAY-17 AT 09:56 BST AT 09:56 BST by MGT The Journal of cell biology 2007 imex curation The yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The Journal of cell biology 2007 imex curation Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk Accepted 2017-MAY-17 AT 09:56 BST AT 09:56 BST by MGT The Journal of cell biology 2007 imex curation The yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The Journal of cell biology 2007 imex curation Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk Accepted 2017-MAY-17 AT 09:56 BST AT 09:56 BST by MGT The Journal of cell biology 2007 imex curation The yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The yeast orthologue of GRASP65 forms a complex with a coiled-coil protein that contributes to ER to Golgi traffic. The Journal of cell biology 2007 imex curation Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip ms participant Identification by mass spectrometry ms participant Behnia R., Barr FA., Flanagan JJ., Barlowe C., Munro S. sean@mrc-lmb.cam.ac.uk Accepted 2017-MAY-17 AT 09:56 BST AT 09:56 BST by MGT The Journal of cell biology 2007 imex curation sec24_yeast Protein transport protein SEC24 SEC24 Abnormal nuclear morphology 1 ANU1 YIL109C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSHHKKRVYPQAQLQYGQNATPLQQPAQFMPPQDPAAAGMSYGQMGMPPQGAVPSMGQQQFLTPAQEQLHQQIDQATTSMNDMHLHNVPLVDPNAYMQPQVPVQMGTPLQQQQQPMAAPAYGQPSAAMGQNMRPMNQLYPIDLLTELPPPITDLTLPPPPLVIPPERMLVPSELSNASPDYIRSTLNAVPKNSSLLKKSKLPFGLVIRPYQHLYDDIDPPPLNEDGLIVRCRRCRSYMNPFVTFIEQGRRWRCNFCRLANDVPMQMDQSDPNDPKSRYDRNEIKCAVMEYMAPKEYTLRQPPPATYCFLIDVSQSSIKSGLLATTINTLLQNLDSIPNHDERTRISILCVDNAIHYFKIPLDSENNEESADQINMMDIADLEEPFLPRPNSMVVSLKACRQNIETLLTKIPQIFQSNLITNFALGPALKSAYHLIGGVGGKIIVVSGTLPNLGIGKLQRRNESGVVNTSKETAQLLSCQDSFYKNFTIDCSKVQITVDLFLASEDYMDVASLSNLSRFTAGQTHFYPGFSGKNPNDIVKFSTEFAKHISMDFCMETVMRARGSTGLRMSRFYGHFFNRSSDLCAFSTMPRDQSYLFEVNVDESIMADYCYVQVAVLLSLNNSQRRIRIITLAMPTTESLAEVYASADQLAIASFYNSKAVEKALNSSLDDARVLINKSVQDILATYKKEIVVSNTAGGAPLRLCANLRMFPLLMHSLTKHMAFRSGIVPSDHRASALNNLESLPLKYLIKNIYPDVYSLHDMADEAGLPVQTEDGEATGTIVLPQPINATSSLFERYGLYLIDNGNELFLWMGGDAVPALVFDVFGTQDIFDIPIGKQEIPVVENSEFNQRVRNIINQLRNHDDVITYQSLYIVRGASLSEPVNHASAREVATLRLWASSTLVEDKILNNESYREFLQIMKARISK 35E2BDD24CC75899 35E2BDD24CC75899 sfb3_yeast SED5-binding protein 3 SFB3 SEC24-related protein 3 Lethal with SEC13 protein 1 LST1 YHR098C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQQNILAASVSALSLDESTVHTGGASSKKSRRPHRAYHNFSSGTVPTLGNSPYTTPQLNQQDGFQQPQAFTPKQFGGFNNGSGSVMSTPVMVSQERFGASEASSPYGQSMLDMTAPQPTSHIVPTQRFEDQAQYLQRSFETCRDSVPPLPTTQFYCVDQGSCDPHLMSLSMYNIPESEHLRAATKLPLGLTIQPFSTLTPNDAEVPTIPLPMDGTPLRCRRCRAYANPKFQFTYDSSVICNICRVKMQVPGEHFAPMGPNGQRSDLNEKSELLHGTVDFLVPSIYNAIQEKELLPLHYVFLIDVSLLANENGSSLAMVEGVRSCIEYISDFQPNCEVAIIVYDNKLRFFNLRPDLDNAQEYIVSELDDVFLPFYNGLFVKPGNSMKIINDTLIKISGYISTDKYSHVPQVCYGSALQAAKLALDTVTGGQGGKIICSLNSLPTIGNGNLSLKRDNAHIAHVKCDNGFYKKLASDFLKSYISLDLYVTNAGFIDMATVGHPVEMTSGILKYYPHFQQETDAFTLVNDMVTNVSNIVGYQALLKVRCSTGLSVEQYYCDSSDNTDHDPIIPVLTRDTTLDVLLKYDSKIKTGTDVHFQTALLYTDIDGVRKVRSINTSGAVSNNIREIFKFINQNPVMRIMIKDVIKTLGDCDFVKIRRLIDDKMVEILTQYRGLVSSNSSTQLILPDSIKTLPAYMLAFEKSELMKPNAQSTRGNERIYDLLKYDSLNSAQLCYKLYPQIVPFHVLLEETDLTFYDANDKLLQINSSSINNLSVRASHSNFINGGCYLIFQGDTIYLWFNENTNRMLLQDLLSVDESLPVSQISLFSGTLPETGTSINQKASNVIKNWQQVVNKSSLPLVLLRPNVDQYYSNVMSQLLCEDKTVNRIESYDNYLVIMHKKIQEKLQKDDFIKVSTAATHENIHQKFVQF 6A028AD6B1A2E031 6A028AD6B1A2E031 sfb2_yeast SED5-binding protein 2 SFB2 ISS1 YNL049C N2505 YNL2505C SEC24-related protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSHHKKRVYPQAQVPYIASMPIVAEQQQSQQQIDQTAYAMGNLQLNNRANSFTQLAQNQQFPGSGKVVNQLYPVDLFTELPPPIRDLSLPPLPITISQDNIVTPSEYSNVPYQYVRSTLKAVPKTNSLLKKTKLPFAIVIRPYLHLQDSDNQVPLNTDGVIVRCRRCRSYMNPFVVFINQGRKWQCNICRFKNDVPFGFDQNLQGAPINRYERNEIKNSVVDYLAPVEYSVREPPPSVYVFLLDVSQNAVKNGLLATSARTILENIEFLPNHDGRTRIAIICVDHSLHYFYVPLDDDYEVSDEDDEESDGEEEDEDEEEEDVDNSETIQMFDIGDLDEPFLPMPSDELVVPLKYCKNNLETLLKKIPEIFQDTHSSKFALGPALKAASNLIKSTGGKVEVISSTLPNTGIGKLKKRSEQGILNTPKESSQLLSCKDSFYKTFTIECNKLQITVDMFLASEDYMDVATLSHLGRFSGGQTHFYPGFNATSLNDVTKFTRELSRHLSMDISMEAVMRVRCSTGLRATSFFGHFFNRSSDLCAFSTMPRDQSYLFGISIEDSLMAEYCYLQVSTLLTLNTGERRIRVMTLALPTSESAREVFASADQLAITDFMTQNAVTKALNSSMYSARDFITKSLEDILNAYKKEISMSNINSVTSLNLCANLRMLPLLMNGLSKHIALRPGVVPSDYRASALNRLETEPLHYLIKSIYPTVYSLHDMPDEVGLPDFEGKTVLPEPINATISLFERYGLYLIDNSAELFLWVGGDAVPELLIDVFNTDTISQIPVGKSELPLLNDSPFNERLRRIIGRIRENNDTITFQSLYIIRGPSINEPANLNSEKDMASLRLWVLSTLVEDKVLNCASYREYLQSMKTSINR 4534CE7E27514379 sec23_yeast Protein transport protein SEC23 SEC23 P9705.14 YPR181C protein protein yeast Saccharomyces cerevisiae Baker's yeast MDFETNEDINGVRFTWNVFPSTRSDANSNVVPVGCLYTPLKEYDELNVAPYNPVVCSGPHCKSILNPYCVIDPRNSSWSCPICNSRNHLPPQYTNLSQENMPLELQSTTIEYITNKPVTVPPIFFFVVDLTSETENLDSLKESIITSLSLLPPNALIGLITYGNVVQLHDLSSETIDRCNVFRGDREYQLEALTEMLTGQKPTGPGGAASHLPNAMNKVTPFSLNRFFLPLEQVEFKLNQLLENLSPDQWSVPAGHRPLRATGSALNIASLLLQGCYKNIPARIILFASGPGTVAPGLIVNSELKDPLRSHHDIDSDHAQHYKKACKFYNQIAQRVAANGHTVDIFAGCYDQIGMSEMKQLTDSTGGVLLLTDAFSTAIFKQSYLRLFAKDEEGYLKMAFNGNMAVKTSKDLKVQGLIGHASAVKKTDANNISESEIGIGATSTWKMASLSPYHSYAIFFEIANTAANSNPMMSAPGSADRPHLAYTQFITTYQHSSGTNRIRVTTVANQLLPFGTPAIAASFDQEAAAVLMARIAVHKAETDDGADVIRWLDRTLIKLCQKYADYNKDDPQSFRLAPNFSLYPQFTYYLRRSQFLSVFNNSPDETAFYRHIFTREDTTNSLIMIQPTLTSFSMEDDPQPVLLDSISVKPNTILLLDTFFFILIYHGEQIAQWRKAGYQDDPQYADFKALLEEPKLEAAELLVDRFPLPRFIDTEAGGSQARFLLSKLNPSDNYQDMARGGSTIVLTDDVSLQNFMTHLQQVAVSGQA 69811913848265FB 69811913848265FB grh1_yeast GRASP65 homolog protein 1 GRH1 YDR517W protein protein yeast Saccharomyces cerevisiae Baker's yeast MFRIAKNLVRTFEQSVQDTLALSQDSSNLDAFFQSIPPNLLSAQLESPVDAVSEGVKHTNVNETLSGLRIVWVDEMQFQLQSFFDYIVGFNDDPVPVVSNQHGFSYPDYRRITSIFNEHCGRTLKVNIWSAKGGTFRDEYISIISKESDDLDDVSLNHDERRPSSGEAHQFQALGFKVQWTPLIASTFTYHILNVNIPDGPAQSAGLIPDEDYIIGCQDGLLATGGETLLQDIVRSRANYDLVLYVYNKVSDCVRPITVHIGPDGRLGCNVGYGFLHRIPTVKHCPQQAQQQGQDDNPVPVPVPVESETAFVPSAFTAPPVPTKKKSKNKKGTQPLAMDDYFNEGRDKSSTAAKSAESDILAPPPQKQSSSD A7CE62DFE4631214 A7CE62DFE4631214 bug1_yeast Binder of USO1 and GRH1 protein 1 BUG1 YDL099W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSEQESDEVKRMKQLEEARKRVEELKKKKNKKNKGKKNKNSSATGSIGSETPDLEGTPGEESTQEETVKANSTKSENNDQNDVDEESEEKEIEQVKSDPSGTTEKDIEEINSTSSNVGKDDAENTKKEEVQEVIKNNNDEQTADAGKTIEPQEEKKIVQTQEGNEPSNTSEAADDLFANDGNEESDFLTTIKKQKEEDELTKLRAENEKLTQENKQLKFLNMENETTVDDLQDQLQEKEDIINGLQNDLQTARDELIAAVEKLKLAEAKAARNTTATPIQFADFNTSSNNLTPSQSVTNSGTQVAHGNNMEVDRVMLNKWRQWNVDMTTWRSIGSGPIMEF E0C5AFBD74062EE9 bug1-grh1-1 1 9 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 10 unspecified role unspecified role neutral component neutral component n-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization Fig. 2A Grh1 and Bug1 colocalize on the cis-Golgi and form a heterooligomeric complex. bug1-grh1-2 3 9 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 2E bug1-grh1-3 3 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 2C, 3C To confirm the interaction between Grh1 and Bug1, HA2-Bug1 was immunoprecipitated, and endogenous Grh1 was found to be enriched in the precipitate bug1-bug1-1 2 10 unspecified role unspecified role bait bait n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role prey prey c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 2D We also found that Bug1-HA3 could be immunoprecipitated with Flag3-Bug1 grh1-sec23-2 4 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association Fig. 4A Interaction was not observed in a MAK3-/- strain grh1-sec23-3 3 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Fig. 4B Interaction was not observed in a MAK3-/- strain