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/ harada-2011-1 Structural studies and the assembly of the heptameric post-translational translocon complex. 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 Harada Y., Li H., Wall J.S., Li H., Lennarz W.J. wlennarz@notes.cc.sunysb.edu J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation harada-2011-5 Structural studies and the assembly of the heptameric post-translational translocon complex. yeast Saccharomyces cerevisiae Baker's yeast bn-page blue native page bn-page anti tag western anti tag western blot anti tag western Harada Y., Li H., Wall J.S., Li H., Lennarz W.J. The highly purified complex with progressively higher concentrations of Triton X-100, ranging from 0.1% to 1%, and then analyzed by BN-PAGE. wlennarz@notes.cc.sunysb.edu J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation harada-2011-3 Structural studies and the assembly of the heptameric post-translational translocon complex. yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip weight silver stain molecular weight estimation by silver staining weight silver stain Harada Y., Li H., Wall J.S., Li H., Lennarz W.J. Microsomes were subjected to glycerol gradient centrifugation at 100,000 × g, then incubated with anti-HA gel. The post-translocon was eluted with HA peptide and further purified by ion-exchange chromatography on Q-sepharose. wlennarz@notes.cc.sunysb.edu J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation harada-2011-6 Structural studies and the assembly of the heptameric post-translational translocon complex. in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Harada Y., Li H., Wall J.S., Li H., Lennarz W.J. Microsomes were subjected to glycerol gradient centrifugation at 100,000 × g, then incubated with anti-HA gel. The post-translocon was eluted with HA peptide and further purified by ion-exchange chromatography on Q-sepharose. wlennarz@notes.cc.sunysb.edu J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation harada-2011-2 Structural studies and the assembly of the heptameric post-translational translocon complex. yeast Saccharomyces cerevisiae Baker's yeast comigration in sds comigration in sds page comigration in sds tag fluorescence tag visualisation by fluorescence tag fluorescence Harada Y., Li H., Wall J.S., Li H., Lennarz W.J. Microsomes weresubjected to glycerol gradient centrifugation at 100,000 × g, then incubated with anti-HA gel. The post-translocon was eluted with HA peptide and further purified by ion-exchange chromatography on Q-sepharose.. The purified post-translocon was denatured, reduced, and subsequently labeled with maleimide containing a fluorescent label. wlennarz@notes.cc.sunysb.edu J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation harada-2011-4 Structural studies and the assembly of the heptameric post-translational translocon complex. in vitro In vitro scanning electron microscopy scanning electron microscopy sem sem predetermined predetermined participant predetermined Harada Y., Li H., Wall J.S., Li H., Lennarz W.J. Microsomes were subjected to glycerol gradient centrifugation at 100,000 × g, then incubated with anti-HA gel. The post-translocon was eluted with HA peptide and further purified by ion-exchange chromatography on Q-sepharose. wlennarz@notes.cc.sunysb.edu J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation sec63_yeast Protein translocation protein SEC63 PTL1 YOR254C SEC63 NPL1 Sec62/63 complex 73 kDa subunit Protein NPL1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPTNYEYDEASETWPSFILTGLLMVVGPMTLLQIYQIFFGANAEDGNSGKSKEFNEEVFKNLNEEYTSDEIKQFRRKFDKNSNKKSKIWSRRNIIIIVGWILVAILLQRINSNDAIKDAATKLFDPYEILGISTSASDRDIKSAYRKLSVKFHPDKLAKGLTPDEKSVMEETYVQITKAYESLTDELVRQNYLKYGHPDGPQSTSHGIALPRFLVDGSASPLLVVCYVALLGLILPYFVSRWWARTQSYTKKGIHNVTASNFVSNLVNYKPSEIVTTDLILHWLSFAHEFKQFFPDLQPTDFEKLLQDHINRRDSGKLNNAKFRIVAKCHSLLHGLLDIACGFRNLDIALGAINTFKCIVQAVPLTPNCQILQLPNVDKEHFITKTGDIHTLGKLFTLEDAKIGEVLGIKDQAKLNETLRVASHIPNLKIIKADFLVPGENQVTPSSTPYISLKVLVRSAKQPLIPTSLIPEENLTEPQDFESQRDPFAMMSKQPLVPYSFAPFFPTKRRGSWCCLVSSQKDGKILQTPIIIEKLSYKNLNDDKDFFDKRIKMDLTKHEKFDINDWEIGTIKIPLGQPAPETVGDFFFRVIVKSTDYFTTDLDITMNMKVRDSPAVEQVEVYSEEDDEYSTDDDETESDDESDASDYTDIDTDTEAEDDESPE sec62_yeast Translocation protein SEC62 SEC62 YPL094C LPG14C Sec62/63 complex 30 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAVGPGSNAGASVNGGSATAIATLLRNHKELKQRQGLFQAKQTDFFRYKRFVRALHSEEYANKSARQPEIYPTIPSNKIEDQLKSREIFIQLIKAQMVIPVKKLHSQECKEHGLKPSKDFPHLIVSNKAQLEADEYFVWNYNPRTYMDYLIVIGVVSIILALVCYPLWPRSMRRGSYYVSLGAFGILAGFFAVAILRLILYVLSLIVYKDVGGFWIFPNLFEDCGVLESFKPLYGFGEKDTYSYKKKLKRMKKKQAKRESNKKKAINEKAEQN sec66_yeast Translocation protein SEC66 SEC71 YBR1232 YBR171W SEC66 HSS1 Sec62/63 complex 31.5 kDa subunit Protein HSS1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSEFNETKFSNNGTFFETEEPIVETKSISVYTPLIYVFILVVSLVMFASSYRKKQAKKISEQPSIFDENDAHDLYFQIKEMSENEKIHEKVLKAALLNRGAESVRRSLKLKELAPQINLLYKNGSIGEDYWKRFETEVKLIELEFKDTLQEAERLQPGWVQLFVMVCKEICFNQALSRRYQSILKRKEVCIKEWELKINNDGRLVN E2A8BB4994A227D4 sec72_yeast Translocation protein SEC72 SEC67 SIM2 L8003.18 YLR292C SEC72 Sec62/63 complex 23 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MVTLEYNANSKLITASDAVVALSTETNIDQINVLTTSLIGETNPNFTPQPNEALSKMIKGLFESGMKNLQQKKLNEALKNVSLAIEMAQRKRAPWEAFAIQLPELHFMLRSKIDLCLILGKHLEALQDLDFLLGTGLIQPDVFVRKADCLLKLRQWEEARATCERGLALAPEDMKLRALLIETARNLAEYNGE sc61a_yeast Protein transport protein SEC61 Sec61 complex subunit SEC61 L3502.5 YLR378C SEC61 Sec61 complex subunit alpha protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSNRVLDLFKPFESFLPEVIAPERKVPYNQKLIWTGVSLLIFLILGQIPLYGIVSSETSDPLYWLRAMLASNRGTLLELGVSPIITSSMIFQFLQGTQLLQIRPESKQDRELFQIAQKVCAIILILGQALVVVMTGNYGAPSDLGLPICLLLIFQLMFASLIVMLLDELLSKGYGLGSGISLFTATNIAEQIFWRAFAPTTVNSGRGKEFEGAVIAFFHLLAVRKDKKRALVEAFYRTNLPNMFQVLMTVAIFLFVLYLQGFRYELPIRSTKVRGQIGIYPIKLFYTSNTPIMLQSALTSNIFLISQILFQKYPTNPLIRLIGVWGIRPGTQGPQMALSGLAYYIQPLMSLSEALLDPIKTIVYITFVLGSCAVFSKTWIEISGTSPRDIAKQFKDQGMVINGKRETSIYRELKKIIPTAAAFGGATIGALSVGSDLLGTLGSGASILMATTTIYGYYEAAAKEGGFTKNLVPGFSDLM sc6b1_yeast Protein transport protein SBH1 Sec61 complex subunit SBH1 YER087BC YER087C-B SBH1 SEB1 Sec61 complex subunit beta protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSPTPPGGQRTLQKRKQGSSQKVAASAPKKNTNSNNSILKIYSDEATGLRVDPLVVLFLAVGFIFSVVALHVISKVAGKLF sc61g_yeast Protein transport protein SSS1 Sec61 complex subunit SSS1 Ssh1 complex subunit SSS1 D4475 YDR086C SSS1 Sec61 complex subunit gamma Ssh1 complex subunit gamma protein protein yeast Saccharomyces cerevisiae Baker's yeast MARASEKGEEKKQSNNQVEKLVEAPVEFVREGTQFLAKCKKPDLKEYTKIVKAVGIGFIAVGIIGYAIKLIHIPIRYVIV sec62-sec63-2 1533290 n/a 1533296 unspecified role unspecified role prey prey c-terminal 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 false n/a 1533297 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 false physical association physical association false false false Fig. 1C, 1D sec62-sec66 1533290 n/a 1533298 unspecified role unspecified role prey prey c-terminal 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 false n/a 1533297 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 false physical association physical association false false false Fig. 1C, 1D sec62-sec72 1533290 n/a 1533299 unspecified role unspecified role prey prey c-terminal 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 false n/a 1533297 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 false physical association physical association false false false Fig. 1C, 1D sec62-sec61 1533290 n/a 1533300 unspecified role unspecified role prey prey c-terminal 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 false n/a 1533297 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 false physical association physical association false false false Fig. 1C, 1D sec62-sbh1 1533290 n/a 1533301 unspecified role unspecified role prey prey c-terminal 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 false n/a 1533297 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 false physical association physical association false false false Fig. 1C, 1D sec62-sss1 1533290 n/a 1533297 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 false n/a 1533302 unspecified role unspecified role prey prey c-terminal 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 false physical association physical association false false false Fig. 1C, 1D sbh1-sec61-4 1533291 n/a 1533299 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533300 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533298 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533302 unspecified role unspecified role neutral component neutral component c-terminal 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 1533297 unspecified role unspecified role neutral component neutral component 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 false n/a 1533296 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533301 unspecified role unspecified role neutral component neutral component c-terminal 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 false physical association physical association false false false Fig. 4B, 4C sbh1-sec61-5 1533291 n/a 1533299 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533298 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533297 unspecified role unspecified role neutral component neutral component 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 false n/a 1533296 unspecified role unspecified role neutral component neutral component c-terminal 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 false physical association physical association false false false Fig. 4B, 4C sbh1-sec61-6 1533291 n/a 1533302 unspecified role unspecified role neutral component neutral component c-terminal 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 1533300 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533301 unspecified role unspecified role neutral component neutral component c-terminal 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 false physical association physical association false false false Fig. 4B, 4C sec62-sec63-3 1533291 n/a 1533299 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533298 unspecified role unspecified role neutral component neutral component c-terminal 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 false n/a 1533296 unspecified role unspecified role neutral component neutral component c-terminal 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 false physical association physical association false false false Fig. 4B, 4C sbh1-sec61-1 1533292 n/a 1533299 unspecified role unspecified role prey prey n/a 1533296 unspecified role unspecified role prey prey n/a 1533300 unspecified role unspecified role prey prey n/a 1533297 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 false n/a 1533302 unspecified role unspecified role prey prey n/a 1533298 unspecified role unspecified role prey prey n/a 1533301 unspecified role unspecified role prey prey association association false false false Fig. 2, 5A sec62-sbh1-1 1533292 n/a 1533297 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 false n/a 1533302 unspecified role unspecified role prey prey n/a 1533298 unspecified role unspecified role prey prey n/a 1533301 unspecified role unspecified role prey prey n/a 1533296 unspecified role unspecified role prey prey c-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1533299 unspecified role unspecified role prey prey n/a 1533300 unspecified role unspecified role prey prey association association false false false Fig. 5A sbh1-sec61-8 1533293 n/a 1533302 unspecified role unspecified role neutral component neutral component n/a 1533301 unspecified role unspecified role neutral component neutral component n/a 1533296 unspecified role unspecified role neutral component neutral component n/a 1533300 unspecified role unspecified role neutral component neutral component n/a 1533299 unspecified role unspecified role neutral component neutral component n/a 1533298 unspecified role unspecified role neutral component neutral component n/a 1533297 unspecified role unspecified role neutral component neutral component 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 false physical association physical association false false false Fig. 6B Complex mass 287 kDa, standard deviation = 42 kDa sbh1-sec61-9 1533293 n/a 1533297 unspecified role unspecified role neutral component neutral component 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 false n/a 1533296 unspecified role unspecified role neutral component neutral component c-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1533301 unspecified role unspecified role neutral component neutral component n/a 1533302 unspecified role unspecified role neutral component neutral component n/a 1533300 unspecified role unspecified role neutral component neutral component n/a 1533299 unspecified role unspecified role neutral component neutral component n/a 1533298 unspecified role unspecified role neutral component neutral component physical association physical association false false false Fig. 6B (lower panel), 7A Complex mass 287 kDa, standard deviation = 42 kDa sec61-sec62 1533294 n/a 1533299 unspecified role unspecified role neutral component neutral component alexa 680 labelled cysteine alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1533296 unspecified role unspecified role neutral component neutral component alexa 680 labelled cysteine alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 n/a 1533298 unspecified role unspecified role neutral component neutral component alexa 680 labelled cysteine alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 n/a 1533300 unspecified role unspecified role neutral component neutral component alexa 680 labelled cysteine alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 n/a 1533297 unspecified role unspecified role neutral component neutral component alexa 680 labelled cysteine alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c_terminus 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 false Stoichiometry: 1.0 n/a 1533302 unspecified role unspecified role neutral component neutral component alexa 680 labelled cysteine alexa label alexa label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 physical association physical association false false false Fig. 2 sbh1-sec61-2 1533295 n/a 1533299 unspecified role unspecified role neutral component neutral component n/a 1533298 unspecified role unspecified role neutral component neutral component n/a 1533297 unspecified role unspecified role neutral component neutral component 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 false n/a 1533300 unspecified role unspecified role neutral component neutral component n/a 1533296 unspecified role unspecified role neutral component neutral component n/a 1533301 unspecified role unspecified role neutral component neutral component n/a 1533302 unspecified role unspecified role neutral component neutral component physical association physical association false false false Fig. 3 Complex mass 287 kDa, standard deviation = 42 kDa