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/ Csg1p and newly identified Csh1p function in mannosylinositol phosphorylceramide synthesis by interacting with Csg2p. Csg1p and newly identified Csh1p function in mannosylinositol phosphorylceramide synthesis by interacting with Csg2p. J. Biol. Chem. (0021-9258) 2003 imex curation Uemura S., Kihara A., Inokuchi J., Igarashi Y. yigarash@pharm.hokudai.ac.jp 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast pull down pull down anti tag western anti tag western blot anti tag western Uemura S., Kihara A., Inokuchi J., Igarashi Y. yigarash@pharm.hokudai.ac.jp Accepted 2009-DEC-10 by JYOTI. 23-Dec-2009: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2003 jyoti Only protein-protein interactions imex curation sur1_yeast Mannosyl phosphorylinositol ceramide synthase SUR1 SUR1 YPL057C BCL21 CSG1 LPE15 protein protein yeast Saccharomyces cerevisiae Baker's yeast MRKELKYLICFNILLLLSIIYYTFDLLTLCIDDTVKDAILEEDLNPDAPPKPQLIPKIIHQTYKTEDIPEHWKEGRQKCLDLHPDYKYILWTDEMAYEFIKEEYPWFLDTFENYKYPIERADAIRYFILSHYGGVYIDLDDGCERKLDPLLAFPAFLRKTSPLGVSNDVMGSVPRHPFFLKALKSLKHYDKYWFIPYMTIMGSTGPLFLSVIWKQYKRWRIPKNGTVRILQPAYYKMHSYSFFSITKGSSWHLDDAKLMKALENHILSCVVTGFIFGFFILYGEFTFYCWLCSKNFSNLTKNWKLNAIKVRFVTILNSLGLRLKLSKSTSDTASATLLARQQKRLRKDSNTNIVLLKSSRKSDVYDLEKNDSSKYSLGNNSS 5A9AAB5F9A4069E3 5A9AAB5F9A4069E3 csg2_yeast Mannosyl phosphorylinositol ceramide synthase regulatory protein CSG2 CLS2 YBR0404 YBR036C CSG2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSTTLLWFSSVIGYVIQTKCLSNIQSKKEISVGPNGTIATPETNGDNGNSSSLTFYLTFMYFASWLLLVPASRLWEKMRPMFVSDSDSNRNSQFDNNNSGSVTNEDVDTFSHVLDDPQPRIPAQQQKQKIISVATFKYVAKLTVLALIMIVADLTYNMALSLSPAFDVALMQNTAIFEIVTLLYGVCGISRKNYVFRNFLIMMNAVIGILIISYTKATCDMLAGKLSVNPNTGELSDPFLFDRLKGALICGLGALIMGPFAVLWNRWFCSNISKNENSAVVLVKQSTHMALIGIIGMVILLPFIPKFPSRESVESISLFYNDKSFWFSLLGSIIFGSLPSLISILELNRKAPAEYLTTCNLGAIIFMGLAEWVCEPTQTTIVRWEVIGYIMLTVSLLVLSVTLGEGKYHH 15780FEBA54E4FD2 csh1_yeast Mannosyl phosphorylinositol ceramide synthase CSH1 YBR161W YBR1212 CSH1 CSG1/SUR1 homolog 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MKKELKILIIANIALLISIIHYTFDLLTLCIDDTSKDALTDEQLNPPNGFNSTFYESPPQLIPKIIHQTYKTNDIPEQWVKGRQKCIDLHPDYTYILWTDEMSDTFIKQEYPWFLDTFRSYEYPIERADAIRYFILSHYGGIYIDLDDGCERRLDPLLKVPAFLRKTSPTGVSNDVMGSVPRHPFFLKVIKSLKHYKKNWYIPYMTIMGSTGPLFISVVWKQYKRWSNTAENGAVRILQPADYKMHNNSFFSISKGSSWHTGDANFMKTLENHILSCVVTGFIFGFFILYGEFTFYTWLCSGPFNNKRYYIQWLSDKFKLHKWKLTSSYKNKEKRRNPTRHEYNSRGKRLRKDSNIPYDSVFLDIEKNHAKFTDLT 34D992039DA0B796 sur1-csg2-1 1 3 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 2 unspecified role unspecified role bait bait c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Although Csg2p-3xHA itself had no affinity to the Ni-NTA column, it was recovered in the eluted fraction when co-expressed with Csg1p-His6-Myc (Fig. 5B). Fig. 5B csh1-csg2-1 1 3 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 4 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Next, to determine the interaction between Csh1p and Csg2p, cell lysates prepared from KHY195 cells bearing pSU8 (CSG2�3�HA) and either pRS423 (vector) or pSU41(CSH1-HIS6-MYC) were subjected to the pull-down assay using the Ni-NTA column. Fig. 5C