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/ Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Cell (0092-8674) 2011 imex curation Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed sde26@cornell.edu jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed Dithiobis[succinimidyl propionate] and Dimethyl 3,3´-dithiobispropionimidate used as cross linking agents. Complexes then immunoprecipitated using anti-HA Abs. sde26@cornell.edu Accepted 2011-APR-19 by JYOTI Cell (0092-8674) 2011 jyoti Only protein-protein interactions imex curation Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Cell (0092-8674) 2011 imex curation Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed sde26@cornell.edu jyoti Only protein-protein interactions in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay anti tag western anti tag western blot anti tag western Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed sde26@cornell.edu Accepted 2011-APR-19 by JYOTI Cell (0092-8674) 2011 jyoti Only protein-protein interactions imex curation Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Cell (0092-8674) 2011 imex curation Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed sde26@cornell.edu jyoti Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed sde26@cornell.edu Accepted 2011-APR-19 by JYOTI Cell (0092-8674) 2011 jyoti Only protein-protein interactions imex curation Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Cell (0092-8674) 2011 imex curation Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed sde26@cornell.edu jyoti Only protein-protein interactions in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Stefan CJ., Manford AG., Baird D., Yamada-Hanff J., Mao Y., Emr SD. I have not added any of the 'in vitro' phosphatase data, as the proteins are in vesicles, not purified, also all colocalisations were with subcellular laocation markers. In several cases, could not work out where proteins were expressed Protein binding alters changes in spectral properties of fluorexcent dye (e.g., emission wavelength maximum, termed “blue shift,” and intensity) when moved from an aqueous environment to a more hydrophobic environment. Assay performed in presence of liposomes (presence of liposomes (PC:PS:PI4P, 3:1:1, 0.1 mM lipid total) sde26@cornell.edu Accepted 2011-APR-19 by JYOTI Cell (0092-8674) 2011 jyoti Only protein-protein interactions imex curation 1-phosphatidyl-1d-myo-inositol 4,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pi3p 1-phosphatidyl-1D-myo-inositol 3-phosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pi4p 1-phosphatidyl-1D-myo-inositol 4-phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) sac1_yeast Phosphatidylinositol-3-phosphatase SAC1 SAC1 RSD1 YKL212W Recessive suppressor of secretory defect Phosphatidylinositol-4-phosphate phosphatase protein protein yeast Saccharomyces cerevisiae Baker's yeast MTGPIVYVQNADGIFFKLAEGKGTNDAVIHLANQDQGVRVLGAEEFPVQGEVVKIASLMGFIKLKLNRYAIIANTVEETGRFNGHVFYRVLQHSIVSTKFNSRIDSEEAEYIKLLELHLKNSTFYFSYTYDLTNSLQRNEKVGPAASWKTADERFFWNHYLTEDLRNFAHQDPRIDSFIQPVIYGYAKTVDAVLNATPIVLGLITRRSIFRAGTRYFRRGVDKDGNVGNFNETEQILLAENPESEKIHVFSFLQTRGSVPIYWAEINNLKYKPNLVLGENSLDATKKHFDQQKELYGDNYLVNLVNQKGHELPVKEGYESVVHALNDPKIHYVYFDFHHECRKMQWHRVKLLIDHLEKLGLSNEDFFHKVIDSNGNTVEIVNEQHSVVRTNCMDCLDRTNVVQSVLAQWVLQKEFESADVVATGSTWEDNAPLLTSYQNLWADNADAVSVAYSGTGALKTDFTRTGKRTRLGAFNDFLNSASRYYQNNWTDGPRQDSYDLFLGGFRPHTASIKSPFPDRRPVYIQLIPMIICAALTVLGATIFFPKDRFTSSKNLLYFAGASIVLALSTKFMFKNGIQFVNWPKLVDVGFLVVHQTHDKEQQFKGLKYAQSPKFSKPDPLKRD 73DB1FA163BC17AE 73DB1FA163BC17AE osh3_yeast Oxysterol-binding protein homolog 3 YHR073W OSH3 protein protein yeast Saccharomyces cerevisiae Baker's yeast METIDIQNRSFVVRWVKCGRGDVINYQIKPLKKSIEVGIYKKLKSSVDDHASAVHIAPDTKTLLDYTTKSLLHKGSSSNIEEHHRRSSQHSHSSSNGSDNKRKERSYSSLSISGIQQQSQEIPLREKLSASGFTLVKRVGNVSGNTMVQGDLEVKDTDYYYAFILDNSSSKNAKKKILFNASVINGDNQSMISTRSTPPARPTALSRTSTQQDMLFRVGQGRYLQGYLLKKRRKRLQGFKKRFFTLDFRYGTLSYYLNDHNQTCRGEIVISLSSVSANKKDKIIIIDSGMEVWVLKATTKENWQSWVDALQTCFDDQFEDKDTSTLEENPDILDDDKEVINKSSPQDHDHLTPTATTKSALSHRQHTQKDMDDIYVPLPSESYATFSMNLRLIQQRLEQCKKDSLSYKPTTLHQRSEGLNGTHSSSSVFTNNRVSSFNHSSSGMTSSDSLASEEVPSNKTYIEHALYNQLADLEVFVSRFVTQGEVLFKDHQILCKKAKDTRVSLTSYLSENDEFFDAEEEISRGVIILPDTEDDINNIVEETPLLGKSDQNEFTKEVQLSGSEQIASSSVESYTTNDENHSRKHLKNRHKNRRRGHPHHQKTKSAQSSTETFTSKDLFALSYPKSVTRRNDIPEAAASPPSLLSFLRKNVGKDLSSIAMPVTSNEPISILQLISETFEYAPLLTKATQRPDPITFVSAFAISFLSIYRDKTRTLRKPFNPLLAETFELIREDMGFRLISEKVSHRPPVFAFFAEHLDWECSYTVTPSQKFWGKSIELNNEGILRLKFKTTGELFEWTQPTTILKNLIAGERYMEPVNEFEVHSSKGDKSHILFDKAGMFSGRSEGFKVSIIPPPSSNRKKETLAGKWTQSLANETTHETIWEVGDLVSNPKKKYGFTKFTANLNEITEIEKGNLPPTDSRLRPDIRAYEEGNVDKAEEWKLKLEQLQRERRNKGQDVEPKYFEKVSKNEWKYITGPKSYWERRKKHDWSDISQLW B1AD1210EFFB9AFE B1AD1210EFFB9AFE scs2_yeast Vesicle-associated membrane protein-associated protein SCS2 YER120W SCS2 VAP homolog 1 Choline sensitivity suppressor protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAVEISPDVLVYKSPLTEQSTEYASISNNSDQTIAFKVKTTAPKFYCVRPNAAVVAPGETIQVQVIFLGLTEEPAADFKCRDKFLVITLPSPYDLNGKAVADVWSDLEAEFKQQAISKKIKVKYLISPDVHPAQNQNIQENKETVEPVVQDSEPKEVPAVVNEKEVPAEPETQPPVQVKKEEVPPVVQKTVPHENEKQTSNSTPAPQNQIKEAATVPAENESSSMGIFILVALLILVLGWFYR 45390E24E31D392E 45390E24E31D392E kes1_yeast Oxysterol-binding protein homolog 4 KES1 OSH4 Protein KES1 LPI3C P2614 YPL145C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQYASSSSWTSFLKSIASFNGDLSSLSAPPFILSPISLTEFSQYWAEHPELFLEPSFINDDNYKEHCLIDPEVESPELARMLAVTKWFISTLKSQYCSRNESLGSEKKPLNPFLGELFVGKWENKEHPEFGETVLLSEQVSHHPPVTAFSIFNDKNKVKLQGYNQIKASFTKSLMLTVKQFGHTMLDIKDESYLVTPPPLHIEGILVASPFVELEGKSYIQSSTGLLCVIEFSGRGYFSGKKNSFKARIYKDSKDSKDKEKALYTISGQWSGSSKIIKANKKEESRLFYDAARIPAEHLNVKPLEEQHPLESRKAWYDVAGAIKLGDFNLIAKTKTELEETQRELRKEEEAKGISWQRRWFKDFDYSVTPEEGALVPEKDDTFLKLASALNLSTKNAPSGTLVGDKEDRKEDLSSIHWRFQRELWDEEKEIVL 360E4A46ABE2D87B osh3-scs2 3 9 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association The GST-Osh31–613 fusion binds Scs2-3xHA from solubilized yeast lysates. Fig. 3B 1_phosphatidy-kes1-1 2 11 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P35844:p.[Arg236Glu;Lys242_Lys243delinsGluGlu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R E certain certain sequence position certain certain certain sequence position certain KK EE Thu Jul 07 12:17:11 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was arg236_lys242_243glu Fri Jul 22 11:15:27 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg236glu,lys242glu,lys243glu' Wed May 02 18:59:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg236glu,lys_lys242-243glu_glu' Tue Jun 30 18:21:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg236Glu;Lys242_Lys243delinsGluGlu]' 6 unspecified role unspecified role bait bait direct interaction direct interaction Supp. Fig. 6B pi4p-kes1 2 7 unspecified role unspecified role bait bait 11 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P35844:p.[Arg236Glu;Lys242_Lys243delinsGluGlu] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R E certain certain sequence position certain certain certain sequence position certain KK EE Thu Jul 07 12:17:17 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was arg236_lys242_243glu Fri Jul 22 11:08:34 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg236glu,lys242glu,lys243glu' Wed May 02 20:03:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg236glu,lys_lys242-243glu_glu' Tue Jun 30 17:39:04 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg236Glu;Lys242_Lys243delinsGluGlu]' direct interaction direct interaction Supp. Fig. 6B 1_phosphatidy-kes1-1 2 11 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait direct interaction direct interaction Supp. Fig. 6B osh3-sac1 1 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 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 Supp. Fig. 7A P13 membrane fractions from cells expressing 3xHA-Osh3 and Sac1-13xmyc or Sac1-13xmyc alone were incubated with cross-linkers, solubilized, immunoprecipitated with anti-HA beads and analyzed by SDS-PAGE and western analysis to detect Osh3-Sac1 complexes. osh7-sac1-3 1 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 7A osh3-sac1-1 4 9 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 8 unspecified role unspecified role neutral component neutral component cys341nbd (4-nitrobenzo-2-oxa-1,3-diazole) fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P32368:p.[Cys392Ser;Cys395Ser] mutation mutation certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:06:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys392ser,cys395ser' Tue Jun 30 20:14:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys392Ser;Cys395Ser]' direct interaction direct interaction Fig. 7B