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/ A second trimeric complex containing homologs of the Sec61p complex functions in protein transport across the ER membrane of S. cerevisiae. A second trimeric complex containing homologs of the Sec61p complex functions in protein transport across the ER membrane of S. cerevisiae. EMBO J. (0261-4189) 1996 imex curation Finke K., Plath K., Panzner S., Prehn S., Rapoport TA., Hartmann E., Sommer T. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom western blot western blot Immuno blot Immuno blot Finke K., Plath K., Panzner S., Prehn S., Rapoport TA., Hartmann E., Sommer T. A digitonin extract was prepared from yeast microsomes, and subjected to chromatography on a Q-Sepharose column. The protein complexes were then bound to SP-Sepharose and eluted with a salt gradient. Accepted 2010-APR-16 by JYOTI. EMBO J. (0261-4189) 1996 jyoti Only protein-protein interactions imex curation A second trimeric complex containing homologs of the Sec61p complex functions in protein transport across the ER membrane of S. cerevisiae. A second trimeric complex containing homologs of the Sec61p complex functions in protein transport across the ER membrane of S. cerevisiae. EMBO J. (0261-4189) 1996 imex curation Finke K., Plath K., Panzner S., Prehn S., Rapoport TA., Hartmann E., Sommer T. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Finke K., Plath K., Panzner S., Prehn S., Rapoport TA., Hartmann E., Sommer T. Yeast cells were solubilized with digitonin, and the extract was centrifuged to obtain a supematant fraction (DE, for digitonin extract) and a ribosome pellet. Membrane proteins associated with the ribosomes were then released by treatment with puromycin and high salt concentrations (referred to as RAMP, for ribosome-associated membrane proteins). Accepted 2010-APR-16 by JYOTI. EMBO J. (0261-4189) 1996 jyoti Only protein-protein interactions imex curation A second trimeric complex containing homologs of the Sec61p complex functions in protein transport across the ER membrane of S. cerevisiae. A second trimeric complex containing homologs of the Sec61p complex functions in protein transport across the ER membrane of S. cerevisiae. EMBO J. (0261-4189) 1996 imex curation Finke K., Plath K., Panzner S., Prehn S., Rapoport TA., Hartmann E., Sommer T. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Finke K., Plath K., Panzner S., Prehn S., Rapoport TA., Hartmann E., Sommer T. Accepted 2010-APR-16 by JYOTI. EMBO J. (0261-4189) 1996 jyoti Only protein-protein interactions imex curation sec62_yeast Translocation protein SEC62 SEC62 YPL094C LPG14C Sec62/63 complex 30 kDa subunit protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAVGPGSNAGASVNGGSATAIATLLRNHKELKQRQGLFQAKQTDFFRYKRFVRALHSEEYANKSARQPEIYPTIPSNKIEDQLKSREIFIQLIKAQMVIPVKKLHSQECKEHGLKPSKDFPHLIVSNKAQLEADEYFVWNYNPRTYMDYLIVIGVVSIILALVCYPLWPRSMRRGSYYVSLGAFGILAGFFAVAILRLILYVLSLIVYKDVGGFWIFPNLFEDCGVLESFKPLYGFGEKDTYSYKKKLKRMKKKQAKRESNKKKAINEKAEQN 2B2BA0DAAF3ACCA1 sc6b2_yeast Protein transport protein SBH2 SBH2 SEB2 Ssh1 complex subunit SBH2 Ssh1 complex subunit beta YER019BC YER019C-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MAASVPPGGQRILQKRRQAQSIKEKQAKQTPTSTRQAGYGGSSSSILKLYTDEANGFRVDSLVVLFLSVGFIFSVIALHLLTKFTHII 5A4F5B9184202E88 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 D5D200DF2DCA81DB 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 AB34F12FB7B3CBD4 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 340F2EA913850D85 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 23B0313EB37744FA 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 E2A8BB4994A227D4 ssh1_yeast Sec sixty-one protein homolog SSH1 Ssh1 complex subunit alpha Ssh1 complex subunit SSH1 YBR2020 YBR283C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSGFRLIDIVKPILPILPEVELPFEKLPFDDKIVYTIFAGLIYLFAQFPLVGLPKATTPNVNDPIYFLRGVFGCEPRTLLEFGLFPNISSGLILQLLAGLKVIKVNFKIQSDRELFQSLTKVFAIVQYVILTNIFIFAGYFGDDLSVVQIGLINFQLVGAGIFTTLLAEVIDKGFGFSSGAMIINTVVIATNLVADTFGVSQIKVGEDDQTEAQGALINLIQGLRSKHKTFIGGIISAFNRDYLPNLTTTIIVLAIAIIVCYLQSVRVELPIRSTRARGTNNVYPIKLLYTGCLSVLFSYTILFYIHIFAFVLIQLVAKNEPTHIICKIMGHYENANNLLAVPTFPLSLLAPPTSFFKGVTQQPLTFITYSAFILVTGIWFADKWQAISGSSARDVALEFKDQGITLMGRREQNVAKELNKVIPIAAVTGASVLSLITVIGESLGLKGKAAGIVVGIAGGFSLLEVITIEYQQSGGQSALNQVLGVPGAM 48267EEA3E0191BD 48267EEA3E0191BD sec61-sbh1-1 3 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Figs. 6A, 7B As expected, antibodies against Sec6lp, Sbhlp or Ssslp brought down all three proteins as subunits of the Sec6lp complex (lanes 1-3). Interaction occurs in SBH2-/- cells sec61-sbh1-2 3 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey association association Fig. 6A sss1-sbh1-1 3 11 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey association association Fig. 6A ssh1-sss1-1 3 8 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association Fig. 6A These data indicate that Sshlp, Sbh2p and Ssslp are contained in a complex. ssh1-sss1-2 3 5 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey association association Figs. 6A, 7A These data indicate that Sshlp, Sbh2p and Ssslp are contained in a complex. Interaction observed in Sbh1-/- cells sec62-sss-1 3 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait association association Fig. 6A sec66-sss-1 3 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey association association Fig. 6A sec61-sbh1-4 1 9 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component physical association physical association Figs. 6C, 7E The trimeric Sec6lp complex eluted mainly in fractions 10 and 11 (arrow 1), as demonstrated by blotting with antibodies directed against its subunits Sec6lp, Sbhlp and Ssslp (Figure 6C). Interaction observed in SEB2-/- cells. ssh1-sss1-3 1 8 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component physical association physical association Figs. 6C, 7D The peak in fraction 12 (arrow 2) is authentic Sshlp complex that contains Sshlp, Sbh2p and Ssslp. Interaction observed in Sbh1-/- cells. ssh1-sss1-4 1 11 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component physical association physical association Fig. 6C, 7E This fraction represents a dimeric complex of Sshlp and Ssslp Interaction occurs in SEB-/- cells sec61-sss1-2 3 7 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Fig. 7A Interaction occurs in SBH1-/- cells sec61-sbh1-5 3 11 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey association association Fig. 7A Interaction occurs in SBH1 -/- cells sec62-sss-2 3 4 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey association association Fig. 6C, 7A, 7B Interaction occurs in SBH1-/- and SEB-/- cells. sec61-sbh1-6 3 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait association association Fig. 7B Interaction occurs in SEB2-/- cells sss1-sbh1-2 3 11 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait association association Fig. 7B Interaction observed in SEB2-/- cells ssh1-sss1-5 3 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association Fig. 7B Interaction occurs in SEB2-/- cells sec61-sss1-3 1 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component physical association physical association Figs. 6C, 7D The trimeric Sec6lp complex eluted mainly in fractions 10 and 11 (arrow 1), as demonstrated by blotting with antibodies directed against its subunits Sec6lp, Sbhlp and Ssslp (Figure 6C). Interaction observed in SBH1-/- cells sec61-sbh1-7 2 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait association association Fig. 8B When the RAMPs were subjected to immunoprecipitation with antibodies against Sec6lp or Sshlp, only the components of the respective trimeric complex were precipitated (Sbhlp and Ssslp, or Sbh2p and Ssslp, respectively; see Figure 8B). ssh1-sss1-6 2 5 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey association association Fig. 8B When the RAMPs were subjected to immunoprecipitation with antibodies against Sec6lp or Sshlp, only the components of the respective trimeric complex were precipitated (Sbhlp and Ssslp, or Sbh2p and Ssslp, respectively; see Figure 8B).