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/ http://www.ebi.ac.uk/ kostelansky-2007-2 Molecular architecture and functional model of the complete yeast ESCRT-I heterotetramer. 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 Kostelansky MS., Schluter C., Tam YY., Lee S., Ghirlando R., Beach B., Conibear E., Hurley JH. hurley@helix.nih.gov Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation kostelansky-2007-1 Molecular architecture and functional model of the complete yeast ESCRT-I heterotetramer. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Kostelansky MS., Schluter C., Tam YY., Lee S., Ghirlando R., Beach B., Conibear E., Hurley JH. hurley@helix.nih.gov Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation kostelansky-2007-3 Molecular architecture and functional model of the complete yeast ESCRT-I heterotetramer. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Kostelansky MS., Schluter C., Tam YY., Lee S., Ghirlando R., Beach B., Conibear E., Hurley JH. Superdex 200 used for molecular seiving. hurley@helix.nih.gov Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation kostelansky-2007-4 Molecular architecture and functional model of the complete yeast ESCRT-I heterotetramer. in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Kostelansky MS., Schluter C., Tam YY., Lee S., Ghirlando R., Beach B., Conibear E., Hurley JH. hurley@helix.nih.gov Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation mvb12_yeast Multivesicular body sorting factor 12 YGR206W G7740 12 kDa multivesicular body sorting factor ESCRT-I complex subunit MVB12 MVB12 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNNNVEELLRRIPLYNKYGKDFPQETVTRFQMPEFKLPALQPTRDLLCPWYEECDNITKVCQLHDSSNKKFDQWYKEQYLSKKPPGIVGNTLLSPSRKDNS E48B085E0FACBAC8 stp22_yeast Suppressor protein STP22 of temperature-sensitive alpha-factor receptor and arginine permease STP22 VPS23 YCL008C YCL8C Vacuolar protein sorting-associated protein 23 ESCRT-I complex subunit VPS23 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSANGKISVPEAVVNWLFKVIQPIYNDGRTTFHDSLALLDNFHSLRPRTRVFTHSDGTPQLLLSIYGTISTGEDGSSPHSIPVIMWVPSMYPVKPPFISINLENFDMNTISSSLPIQEYIDSNGWIALPILHCWDPAAMNLIMVVQELMSLLHEPPQDQAPSLPPKPNTQLQQEQNTPPLPPKPKSPHLKPPLPPPPPPQPASNALDLMDMDNTDISPTNHHEMLQNLQTVVNELYREDVDYVADKILTRQTVMQESIARFHEIIAIDKNHLRAVEQAIEQTMHSLNAQIDVLTANRAKVQQFSSTSHVDDEDVNSIAVAKTDGLNQLYNLVAQDYALTDTIECLSRMLHRGTIPLDTFVKQGRELARQQFLVRWHIQRITSPLS FEDE3BE79F063BCE srn2_yeast Protein SRN2 SRN2 ESCRT-I complex subunit VPS37 SRN10 Vacuolar protein sorting-associated protein 37 VPS37 L2958 L9233.5 YLR119W protein protein yeast Saccharomyces cerevisiae Baker's yeast MKVKATKLRIKQRRKNKGLNISRLDIIRAEMDVVPSPGLPEKVNEKSKNIPLPEGINLLSSKEIIDLIQTHRHQLELYVTKFNPLTDFAGKIHAFRDQFKQLEENFEDLHEQKDKVQALLENCRILESKYVASWQDYHSEFSKKYGDIALKKKLEQNTKKLDEESSQLETTTRSIDSADDLDQFIKNYLDIRTQYHLRREKLATWDKQGNLKY 4E99806CED03E345 vps28_yeast Vacuolar protein sorting-associated protein 28 VPS28 VPT28 YPL065W LPE5W ESCRT-I complex subunit VPS28 protein protein yeast Saccharomyces cerevisiae Baker's yeast MQKHNIKLNQNQDISQLFHDEVPLFDNSITSKDKEVIETLSEIYSIVITLDHVEKAYLKDSIDDTQYTNTVDKLLKQFKVYLNSQNKEEINKHFQSIEAFCDTYNITASNAITRLERGIPITAEHAISTTTSAPSGDNKQSSSSDKKFNAKYVAEATGNFITVMDALKLNYNAKDQLHPLLAELLISINRVTRDDFENRSKLIDWIVRINKLSIGDTLTETQIRELLFDLELAYKSFYALLD DE091D3F60C9D132 mvb12 vps23 652965 N/A 652969 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P42939:p.[Leu47Asp;Ile57Asp] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 652970 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 false P25604:p.Met254Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 physical association physical association false false false 1E, 5D, S5D The stoichiometry was determined as percent of the protein in the cell lysate. The copurification of Mvb12 and Vps23 required Vps37. vps37-vps23-1 652965 N/A 652971 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 false N/A 652970 unspecified role unspecified role bait bait P25604:p.Met254Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Stoichiometry: 1.0 physical association physical association false false false 5D vps37-mvb12 652965 N/A 652969 unspecified role unspecified role prey prey region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P42939:p.[Leu47Asp;Ile57Asp] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false N/A 652971 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 5E mvb12 vps23-1 652965 N/A 652970 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 false Stoichiometry: 1.0 N/A 652969 unspecified role unspecified role prey prey region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P42939:p.[Leu47Asp;Ile57Asp] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 physical association physical association false false false 5E stp22-srn2-1 652966 N/A 652972 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652971 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652970 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652969 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false working 23.3 percent, free 31.5 percent 2.70 A 2, 3 Table S1 stp22-srn2-2 652967 N/A 652971 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652972 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652970 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652969 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false 4B ESCRT-I complex consisting of intact Vps23, Vps28, Vps37, and Mvb12 runs on gel filtration chromatography with an apparent molecular weight of 340 kDa stp22-srn2-3 652968 N/A 652971 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652970 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652969 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 652972 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false 4B ESCRT-I complex has a molecular mass of 117 Kda.