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/ Structural and functional organization of the ESCRT-I trafficking complex. Structural and functional organization of the ESCRT-I trafficking complex. Cell (0092-8674) 2006 imex curation Kostelansky MS., Sun J., Lee S., Kim J., Ghirlando R., Hierro A., Emr SD., Hurley JH. hurley@helix.nih.gov 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 4 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 Accepted 2007-FEB-6 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2006 Only protein-protein interactions 4 imex curation Structural and functional organization of the ESCRT-I trafficking complex. Structural and functional organization of the ESCRT-I trafficking complex. Cell (0092-8674) 2006 imex curation Kostelansky MS., Sun J., Lee S., Kim J., Ghirlando R., Hierro A., Emr SD., Hurley JH. hurley@helix.nih.gov 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 4 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. Interactions from figure 6 not included since the organism of the proteins forming the ESCRT II not known. Auhtor contacted - no response. 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 Accepted 2007-FEB-6 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2006 Only protein-protein interactions 4 imex curation Structural and functional organization of the ESCRT-I trafficking complex. Structural and functional organization of the ESCRT-I trafficking complex. Cell (0092-8674) 2006 imex curation Kostelansky MS., Sun J., Lee S., Kim J., Ghirlando R., Hierro A., Emr SD., Hurley JH. hurley@helix.nih.gov 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 4 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 Accepted 2007-FEB-6 by DAVET. 02-MAR-2006: Contacted by JYOTI. 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 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 DE091D3F60C9D132 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 FEDE3BE79F063BCE vps28-stp22 3 4 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 [EBI-1178318]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 214-214, sequence length 213 [EBI-1178318]132-214 ecoli Escherichia coli (strain K12) 6 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 ecoli Escherichia coli (strain K12) 5 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 ecoli Escherichia coli (strain K12) physical association physical association 4, table S2 2.8A working 20.3, free 25.6 The crystal structures of a Vps23:Vps28 core subcomplex and the Vps23:Vps28:Vps37 core were solved at 2.1 and 2.8 � resolution. vps28-stp22-1 3 5 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 ecoli Escherichia coli (strain K12) 6 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction 2, table S2 2.1A working 24.1, free 26.1 The crystal structures of a Vps23:Vps28 core subcomplex and the Vps23:Vps28:Vps37 core were solved at 2.1 and 2.8 � resolution. vps28-stp22-2 2 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 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 ecoli Escherichia coli (strain K12) physical association physical association 1A, 1B This study commenced with the expression and characterization of a recombinant form of the yeast ESCRT-I complex, referred to here as EI[Delta]UEV-A, in which the UEV domain of Vps23, residues 1�161, was deleted (Fig. 1A). The above complex yeilded a predominant peak with apparent MW of 190 kDa. Details of experimental procedure not provided hence data from table S3 and 5B not incorporated. No response from authors. vps28-stp22-3 1 6 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association 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. Sedimentation velocity studies of the EI[Delta]UEV-A complex indicated the presence of two species having corrected sedimentation coefficients of approximately 3.6 and 4.8 S (Fig. 1C). Modeling the equilibrium data to a monomerdimer mixture yielded excellent fits a dissociation constant constant Kd of 110-160 micromoles.