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-2006-1 Structural and functional organization of the ESCRT-I trafficking complex. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Kostelansky MS., Sun J., Lee S., Kim J., Ghirlando R., Hierro A., Emr SD., Hurley JH. hurley@helix.nih.gov Cell (0092-8674) 2006 Only protein-protein interactions 4 imex curation kostelansky-2006-2 Structural and functional organization of the ESCRT-I trafficking complex. 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., Sun J., Lee S., Kim J., Ghirlando R., Hierro A., Emr SD., Hurley JH. Gel filteration followed by MS and protein sequencing were used to identify the core of the ESCRT I complex. The complex was analysed by Abs 280 of the proteins. hurley@helix.nih.gov Cell (0092-8674) 2006 Only protein-protein interactions 4 imex curation kostelansky-2006-3 Structural and functional organization of the ESCRT-I trafficking complex. in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Kostelansky MS., Sun J., Lee S., Kim J., Ghirlando R., Hierro A., Emr SD., Hurley JH. Gel filtered ESCRT I complex with deleted VPS23 was used for sedimentation analysis. Sedimentation equilibrium centrifugation and Sedimentation velocity centrifugation were both used to determine the complex properties. Abs 280nm was used to determine the proteins in the fractions. hurley@helix.nih.gov Cell (0092-8674) 2006 Only protein-protein interactions 4 imex curation 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 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 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 vps28-stp22 492249 N/A 492252 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 492253 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 false ecoli Escherichia coli (strain K12) N/A 492254 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false 4, table S2 2.8A working 20.3, free 25.6 vps28-stp22-1 492249 N/A 492254 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 false ecoli Escherichia coli (strain K12) N/A 492253 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 2, table S2 2.1A working 24.1, free 26.1 vps28-stp22-2 492250 N/A 492254 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) N/A 492252 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) N/A 492253 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false 1A, 1B The above complex yeilded a predominant peak with apparent MW of 190 kDa. vps28-stp22-3 492251 N/A 492253 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 492254 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 492252 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false 1C, 1D, 1F The above complex exist as two species having sedimentation coefficients of 3.6 and 4.8S. Monomeric 1:1:1 ternary complex with Molecular weight of 78 kDa is the major species with a dimeric state also observed. Modeling the equilibrium data to a monomerdimer mixture yielded excellent fits a dissociation constant constant Kd of 110-160 micromoles.