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/ Tsc3p is an 80-amino acid protein associated with serine palmitoyltransferase and required for optimal enzyme activity. Tsc3p is an 80-amino acid protein associated with serine palmitoyltransferase and required for optimal enzyme activity. J. Biol. Chem. (0021-9258) 2000 imex curation Gable K., Slife H., Bacikova D., Monaghan E., Dunn TM. Coimmunoprecipitations performed in microsomes solubilized with 2 mM sucrose monolaurate Tdunn@usuhs.mil 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Gable K., Slife H., Bacikova D., Monaghan E., Dunn TM. Coimmunoprecipitations performed in microsomes solubilized with 2 mM sucrose monolaurate Tdunn@usuhs.mil Accepted 2009-FEB-21 by DAVET. 09-April-2009: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2000 davet Only protein-protein interactions imex curation Tsc3p is an 80-amino acid protein associated with serine palmitoyltransferase and required for optimal enzyme activity. Tsc3p is an 80-amino acid protein associated with serine palmitoyltransferase and required for optimal enzyme activity. J. Biol. Chem. (0021-9258) 2000 imex curation Gable K., Slife H., Bacikova D., Monaghan E., Dunn TM. Coimmunoprecipitations performed in microsomes solubilized with 2 mM sucrose monolaurate Tdunn@usuhs.mil 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Gable K., Slife H., Bacikova D., Monaghan E., Dunn TM. Although Lcb1 and Lcb2 were separately shown to bind to Tsc3, the author has shown that the two proteins are unstable in the absence of their partner, so infers that all 3 proteins must be present in each coimmunoprecipitation. The data has been annotated to reflect this. Coimmunoprecipitations performed in microsomes solubilized with 2 mM sucrose monolaurate Tdunn@usuhs.mil Accepted 2009-FEB-21 by DAVET. 09-April-2009: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2000 davet Only protein-protein interactions imex curation Tsc3p is an 80-amino acid protein associated with serine palmitoyltransferase and required for optimal enzyme activity. Tsc3p is an 80-amino acid protein associated with serine palmitoyltransferase and required for optimal enzyme activity. J. Biol. Chem. (0021-9258) 2000 imex curation Gable K., Slife H., Bacikova D., Monaghan E., Dunn TM. Coimmunoprecipitations performed in microsomes solubilized with 2 mM sucrose monolaurate Tdunn@usuhs.mil 09-April-2009: Contacted by JYOTI. davet 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 Gable K., Slife H., Bacikova D., Monaghan E., Dunn TM. Coimmunoprecipitations performed in microsomes solubilized with 2 mM sucrose monolaurate Tdunn@usuhs.mil Accepted 2009-FEB-21 by DAVET. 09-April-2009: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2000 davet Only protein-protein interactions imex curation lcb2_yeast Serine palmitoyltransferase 2 LCB2 SCS1 YDR062W YD9609.16 D4246 TSC1 Long chain base biosynthesis protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSTPANYTRVPLCEPEELPDDIQKENEYGTLDSPGHLYQVKSRHGKPLPEPVVDTPPYYISLLTYLNYLILIILGHVHDFLGMTFQKNKHLDLLEHDGLAPWFSNFESFYVRRIKMRIDDCFSRPTTGVPGRFIRCIDRISHNINEYFTYSGAVYPCMNLSSYNYLGFAQSKGQCTDAALESVDKYSIQSGGPRAQIGTTDLHIKAEKLVARFIGKEDALVFSMGYGTNANLFNAFLDKKCLVISDELNHTSIRTGVRLSGAAVRTFKHGDMVGLEKLIREQIVLGQPKTNRPWKKILICAEGLFSMEGTLCNLPKLVELKKKYKCYLFIDEAHSIGAMGPTGRGVCEIFGVDPKDVDILMGTFTKSFGAAGGYIAADQWIIDRLRLDLTTVSYSESMPAPVLAQTISSLQTISGEICPGQGTERLQRIAFNSRYLRLALQRLGFIVYGVADSPVIPLLLYCPSKMPAFSRMMLQRRIAVVVVAYPATPLIESRVRFCMSASLTKEDIDYLLRHVSEVGDKLNLKSNSGKSSYDGKRQRWDIEEVIRRTPEDCKDDKYFVN 06DF4A709CC9F29B 06DF4A709CC9F29B lcb1_yeast Serine palmitoyltransferase 1 YMR296C LCB1 END8 TSC2 Long chain base biosynthesis protein 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MAHIPEVLPKSIPIPAFIVTTSSYLWYYFNLVLTQIPGGQFIVSYIKKSHHDDPYRTTVEIGLILYGIIYYLSKPQQKKSLQAQKPNLSPQEIDALIEDWEPEPLVDPSATDEQSWRVAKTPVTMEMPIQNHITITRNNLQEKYTNVFNLASNNFLQLSATEPVKEVVKTTIKNYGVGACGPAGFYGNQDVHYTLEYDLAQFFGTQGSVLYGQDFCAAPSVLPAFTKRGDVIVADDQVSLPVQNALQLSRSTVYYFNHNDMNSLECLLNELTEQEKLEKLPAIPRKFIVTEGIFHNSGDLAPLPELTKLKNKYKFRLFVDETFSIGVLGATGRGLSEHFNMDRATAIDITVGSMATALGSTGGFVLGDSVMCLHQRIGSNAYCFSACLPAYTVTSVSKVLKLMDSNNDAVQTLQKLSKSLHDSFASDDSLRSYVIVTSSPVSAVLHLQLTPAYRSRKFGYTCEQLFETMSALQKKSQTNKFIEPYEEEEKFLQSIVDHALINYNVLITRNTIVLKQETLPIVPSLKICCNAAMSPEELKNACESVKQSILACCQESNK 9F7F93E4B2C70FDB 9F7F93E4B2C70FDB tsc3_yeast Serine palmitoyltransferase-regulating protein TSC3 TSC3 Temperature-sensitive CSG2-mutant suppressor protein 3 YBR058C-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MTQHKSSMVYIPTTKEAKRRNGKSEGILNTIEEVVEKLYWTYYIHLPFYLMASFDSFFLHVFFLTIFSLSFFGILKYCFL F23EE19603FEC87E F23EE19603FEC87E lcb1-lcb2-1 3 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association The rabbit polyclonal antibodies to either Lcb1p or Lcb2p coimmunoprecipitate both Lcb1p and Lcb2p, indicating that the two proteins are associated (Fig. 5A). Figs. 5A and 5B Interaction does not require the presence of Tsc3 lcb1-lcb2-2 3 4 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association The rabbit polyclonal antibodies to either Lcb1p or Lcb2p coimmunoprecipitate both Lcb1p and Lcb2p, indicating that the two proteins are associated (Fig. 5A). Figs. 5A and 5B Interaction does not require the presence of Tsc3 lcb1-lcb2-3 3 5 unspecified role unspecified role prey prey region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait physical association physical association The Lcb1p-GST fusion protein restores SPT activity (data not shown) and coimmunoprecipitates with Lcb2p (Fig. 5A, lanes 6 and 7). Fig. 5A Tagged protein was expressed in cells lacking lcb1 lcb1-lcb2-4 3 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The Lcb1p-GST fusion protein restores SPT activity (data not shown) and coimmunoprecipitates with Lcb2p (Fig. 5A, lanes 6 and 7). Fig. 5A Tagged protein was expressed in cells lacking lcb1 lcb1-lcb2-5 2 4 unspecified role unspecified role bait bait 6 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 5 unspecified role unspecified role prey prey association association The rabbit polyclonal antibodies to either Lcb1p or Lcb2p coimmunoprecipitated Tsc3p-HA along with Lcb1p/Lcb2p (Fig. 6C) Fig. 6C Tsc3 is lost from this protein complex at high salt concentrations lcb1-lcb2-6 2 5 unspecified role unspecified role bait bait 6 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 4 unspecified role unspecified role prey prey association association The rabbit polyclonal antibodies to either Lcb1p or Lcb2p coimmunoprecipitated Tsc3p-HA along with Lcb1p/Lcb2p (Fig. 6C) Fig. 6C Tsc3 is lost from this protein complex at high salt concentrations lcb1-lcb2-7 1 6 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 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role prey prey association association or conversely, the monoclonal anti-HA antibodies coimmunoprecipitated Lcb1p and Lcb2p along with Tsc3p-HA (Fig. 6D). Fig. 6D