UniProt UniProt http://www.uniprot.org Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. mbard@iupui.edu 07-Sept-2007: Contacted by JYOTI. orchard yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. mbard@iupui.edu Accepted 2007-AUG-30 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. mbard@iupui.edu 07-Sept-2007: Contacted by JYOTI. orchard yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast cosedimentation cosedimentation anti tag western anti tag western blot anti tag western Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. Samples were subjected to two rounds of sucrose gradient centrifugation; (10-60%) and (30-60%). mbard@iupui.edu Accepted 2007-AUG-30 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. mbard@iupui.edu 07-Sept-2007: Contacted by JYOTI. orchard yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip antibody detection identification by antibody antibody detection Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. Solubilized membrane protein of cells either transformed with ERG27-HA and Myc-ERG28 or ERG26-HA and Myc-ERG28 were coimmunoprecipitated with mouse anti-HA matrix beads. mbard@iupui.edu Accepted 2007-AUG-30 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Protein-protein interactions among C-4 demethylation enzymes involved in yeast sterol biosynthesis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. mbard@iupui.edu 07-Sept-2007: Contacted by JYOTI. orchard 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 Mo C., Valachovic M., Randall SK., Nickels JT., Bard M. mbard@iupui.edu Accepted 2007-AUG-30 by ORCHARD. 07-Sept-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard msmo_yeast Methylsterol monooxygenase ERG25 FET6 C-4 methylsterol oxidase Sterol-C4-methyl oxidase YGR060W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAVFNNATLSGLVQASTYSQTLQNVAHYQPQLNFMEKYWAAWYSYMNNDVLATGLMFFLLHEFMYFFRCLPWFIIDQIPYFRRWKLQPTKIPSAKEQLYCLKSVLLSHFLVEAIPIWTFHPMCEKLGITVEVPFPSLKTMALEIGLFFVLEDTWHYWAHRLFHYGVFYKYIHKQHHRYAAPFGLSAEYAHPAETLSLGFGTVGMPILYVMYTGKLHLFTLCVWITLRLFQAVDSHSGYDFPWSLNKIMPFWAGAEHHDLHHHYFIGNYASSFRWWDYCLDTESGPEAKASREERMKKRAENNAQKKTN E0422C416DD17794 erg26_yeast Sterol-4-alpha-carboxylate 3-dehydrogenase, decarboxylating ERG26 C-3 sterol dehydrogenase C-4 decarboxylase NAD(P)-dependent cholesterol dehydrogenase YGL001C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSKIDSVLIIGGSGFLGLHLIQQFFDINPKPDIHIFDVRDLPEKLSKQFTFNVDDIKFHKGDLTSPDDMENAINESKANVVVHCASPMHGQNPDIYDIVNVKGTRNVIDMCKKCGVNILVYTSSAGVIFNGQDVHNADETWPIPEVPMDAYNETKAIAEDMVLKANDPSSDFYTVALRPAGIFGPGDRQLVPGLRQVAKLGQSKFQIGDNNNLFDWTYAGNVADAHVLAAQKLLDPKTRTAVSGETFFITNDTPTYFWALARTVWKADGHIDKHVIVLKRPVAICAGYLSEWVSKMLGKEPGLTPFRVKIVCAYRYHNIAKAKKLLGYTPRVGIEEGINKTLAWMDEGL 87CA8FF8D64B0327 87CA8FF8D64B0327 erg27_yeast 3-keto-steroid reductase ERG27 YLR100W protein protein yeast Saccharomyces cerevisiae Baker's yeast MNRKVAIVTGTNSNLGLNIVFRLIETEDTNVRLTIVVTSRTLPRVQEVINQIKDFYNKSGRVEDLEIDFDYLLVDFTNMVSVLNAYYDINKKYRAINYLFVNAAQGIFDGIDWIGAVKEVFTNPLEAVTNPTYKIQLVGVKSKDDMGLIFQANVFGPYYFISKILPQLTRGKAYIVWISSIMSDPKYLSLNDIELLKTNASYEGSKRLVDLLHLATYKDLKKLGINQYVVQPGIFTSHSFSEYLNFFTYFGMLCLFYLARLLGSPWHNIDGYKAANAPVYVTRLANPNFEKQDVKYGSATSRDGMPYIKTQEIDPTGMSDVFAYIQKKKLEWDEKLKDQIVETRTPI D9A0589C049D8C49 D9A0589C049D8C49 erg28_yeast Ergosterol biosynthetic protein 28 YER044C ERG28 BUD18 protein protein yeast Saccharomyces cerevisiae Baker's yeast MFSLQDVITTTKTTLAAMPKGYLPKWLLFISIVSVFNSIQTYVSGLELTRKVYERKPTETTHLSARTFGTWTFISCVIRFYGAMYLNEPHIFELVFMSYMVALFHFGSELLIFRTCKLGKGFMGPLVVSTTSLVWMYKQREYYTGVAW F1BD39AFA180E367 F1BD39AFA180E367 erg28-erg27-1 1 7 unspecified role unspecified role prey prey region 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 8 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 5B It was found that Erg28p and Erg27p reciprocally coimmunoprecipitated (Fig. 5B); erg27-erg28-1 1 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 7 unspecified role unspecified role bait bait region 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 physical association physical association 5B It was found that Erg28p and Erg27p reciprocally coimmunoprecipitated (Fig. 5B); erg25-erg27-1 4 5 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey region 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 physical association physical association 5D Erg25p also coimmunoprecipitated with Erg27p and Erg28p by using rabbit anti-Erg25p (Fig. 5D). erg25-erg28-2 4 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait physical association physical association 5D Erg25p also coimmunoprecipitated with Erg27p and Erg28p by using rabbit anti-Erg25p (Fig. 5D). erg27-erg28-2 2 7 unspecified role unspecified role neutral component neutral component region 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 8 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 6A Erg26p/Erg28p and Erg27p/Erg28p Comigrate in the Same Complex. erg26-erg28-1 2 6 unspecified role unspecified role neutral component neutral component region 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 8 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association 6B Erg26p/Erg28p and Erg27p/Erg28p Comigrate in the Same Complex. erg27-erg28-erg25-1 3 7 unspecified role unspecified role bait bait region 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 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role prey prey association association 8.1 erg26-erg25-erg28-1 3 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role bait bait region 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 5 unspecified role unspecified role prey prey association association 8.2