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/ Functional reconstitution of ESCRT-III assembly and disassembly. Functional reconstitution of ESCRT-III assembly and disassembly. Cell (0092-8674) 2009 imex curation Saksena S., Wahlman J., Teis D., Johnson AE., Emr SD. I have not attempted to capture the data in Fig. 6A where up to 6 components are present, as it does not appear to me to clear what is binding to what - you can't tell if its one big complex or several small ones. ajohnson@medicine.tamhsc.edu, sde26@cornell.edu 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 6 in vitro In vitro fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret predetermined predetermined participant predetermined Saksena S., Wahlman J., Teis D., Johnson AE., Emr SD. I have not attempted to capture the data in Fig. 6A where up to 6 components are present, as it does not appear to me to clear what is binding to what - you can't tell if its one big complex or several small ones. ajohnson@medicine.tamhsc.edu, sde26@cornell.edu Accepted 2009-MAR-17 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation Functional reconstitution of ESCRT-III assembly and disassembly. Functional reconstitution of ESCRT-III assembly and disassembly. Cell (0092-8674) 2009 imex curation Saksena S., Wahlman J., Teis D., Johnson AE., Emr SD. I have not attempted to capture the data in Fig. 6A where up to 6 components are present, as it does not appear to me to clear what is binding to what - you can't tell if its one big complex or several small ones. ajohnson@medicine.tamhsc.edu, sde26@cornell.edu 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 6 in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Saksena S., Wahlman J., Teis D., Johnson AE., Emr SD. I have not attempted to capture the data in Fig. 6A where up to 6 components are present, as it does not appear to me to clear what is binding to what - you can't tell if its one big complex or several small ones. ajohnson@medicine.tamhsc.edu, sde26@cornell.edu Accepted 2009-MAR-17 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation Functional reconstitution of ESCRT-III assembly and disassembly. Functional reconstitution of ESCRT-III assembly and disassembly. Cell (0092-8674) 2009 imex curation Saksena S., Wahlman J., Teis D., Johnson AE., Emr SD. I have not attempted to capture the data in Fig. 6A where up to 6 components are present, as it does not appear to me to clear what is binding to what - you can't tell if its one big complex or several small ones. ajohnson@medicine.tamhsc.edu, sde26@cornell.edu 09-April-2009: Contacted by JYOTI. davet Only protein-protein interactions 6 in vitro In vitro density sedimentation cosedimentation through density gradient density sedimentation weight by staining molecular weight estimation by staining weight by staining Saksena S., Wahlman J., Teis D., Johnson AE., Emr SD. I have not attempted to capture the data in Fig. 6A where up to 6 components are present, as it does not appear to me to clear what is binding to what - you can't tell if its one big complex or several small ones. 10%�40% Glycerol density gradient ajohnson@medicine.tamhsc.edu, sde26@cornell.edu Accepted 2009-MAR-17 by DAVET. 09-April-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation vps25_yeast Vacuolar protein-sorting-associated protein 25 YJR102C J1957 VPS25 VPT25 ESCRT-II complex subunit VPS25 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSALPPVYSFPPLYTRQPNSLTRRQQISTWIDIISQYCKTKKIWYMSVDGTVINDNELDSGSTDNDDSKKISKNLFNNEDIQRSVSQVFIDEIWSQMTKEGKCLPIDQSGRRSSNTTTTRYFILWKSLDSWASLILQWFEDSGKLNQVITLYELSEGDETVNWEFHRMPESLLYYCLKPLCDRNRATMLKDENDKVIAIKVV 6A4545A8BC772CA2 6A4545A8BC772CA2 snf7_yeast Vacuolar-sorting protein SNF7 SNF7 VPS32 YLR025W DID1 Vacuolar protein-sorting-associated protein 32 DOA4-independent degradation protein 1 Sucrose nonfermenting protein 7 protein protein yeast Saccharomyces cerevisiae Baker's yeast MWSSLFGWTSSNAKNKESPTKAIVRLREHINLLSKKQSHLRTQITNQENEARIFLTKGNKVMAKNALKKKKTIEQLLSKVEGTMESMEQQLFSIESANLNLETMRAMQEGAKAMKTIHSGLDIDKVDETMDEIREQVELGDEISDAISRPLITGANEVDEDELDEELDMLAQENANQETSKIVNNNVNAAPISENKVSLPSVPSNKIKQSENSVKDGEEEEDEEDEDEKALRELQAEMGL 5241A18BB181F0C1 5241A18BB181F0C1 vps20_yeast Vacuolar protein sorting-associated protein 20 ASI10 CHM6 VPT20 YM9582.02C YMR077C VPS20 Vacuolar protein-targeting protein 20 Amino acid sensor-independent protein 10 Charged multivesicular body protein 6 ESCRT-III complex subunit VPS20 protein protein yeast Saccharomyces cerevisiae Baker's yeast MGQKSSKVHITKTDRAILEVKRSKDEIHKFTRRTDNLILVEKSQLKDLIRKNPENYKSNMKVRFLLKRIHYQEHLLQQASDQLINLENMVSTLEFKMVEKQFINGLKNGNEILKKLNKEFSNVDELMDDVQDQIAYQNEINETLSRSLVGTSNYEDDLDKELDALESELNPEKMNNAKVANMPSTEGLPSLPQGEQTEQKEREEFATEERSDTKEPLALLS CDC88BC4ADA3D602 CDC88BC4ADA3D602 vps20-vps25-1 2 4 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component Q04272:p.Lys61Cys mutation mutation certain certain sequence position certain certain certain sequence position certain K C Wed May 02 19:37:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys61cys' Tue Jun 30 17:03:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys61Cys' ecoli Escherichia coli (strain K12) direct interaction direct interaction A fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye was covalently attached to the Cys mutated into the protein sequence. However, when Vps25 was added to membrane-bound Vps2061-NBD, a large additional increase of NBD emission intensity was observed that was Vps25 dependent (Figures 2D-2F). Vps20 must be membrane-bound for interaction to occur. Figs. 2D-2F, Supp. Fig. 4 vps20-snf7-1 2 5 unspecified role unspecified role neutral component neutral component P39929:p.Gly58Cys mutation mutation certain certain sequence position certain certain certain sequence position certain G C Wed May 02 18:09:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly58cys' Tue Jun 30 19:39:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly58Cys' ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction A fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye was covalently attached to the Cys mutated into the protein sequence. Interestingly, the spectral data indicate that Vps20 binds similarly to both free and membrane-bound Snf7. Vps20 binds to both free and membrane-bound Snf7. Fig. 4, Supp. Fig. 7 A probe at postion 200 of SNF7 was sensitive to membrane binding but not VPS20 binding. snf7-1 1 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept glu-81 rhodamine fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role fluorescence donor fluorescence donor P39929:p.Glu81Cys mutation mutation certain certain sequence position certain certain certain sequence position certain E C Wed May 02 18:09:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu81cys' Tue Jun 30 19:38:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu81Cys' cys-81 fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction The association of Snf7 into polymeric assemblies was detected directly through the use of a variation of the fluorescence resonance energy transfer (FRET) technique involving two samples of Snf7 E81C, one labeled with an NBD donor dye and the other with a Rh acceptor dye. Figs. 5A, 5B, Supp. Table 1 Both membranes and Vps20 are required for maximal association of Snf7 molecules. Vps20 (Q04272) and liposomes A fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye was covalently attached to the Cys mutated into the protein sequence. snf7-2 1 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept glu-81 rhodamine fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role fluorescence donor fluorescence donor cys-21 fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain P39929:p.Lys21Cys mutation mutation certain certain sequence position certain certain certain sequence position certain K C Wed May 02 14:43:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys21cys' Tue Jun 30 17:25:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys21Cys' ecoli Escherichia coli (strain K12) direct interaction direct interaction The association of Snf7 into polymeric assemblies was detected directly through the use of a variation of the fluorescence resonance energy transfer (FRET) technique involving two samples of Snf7 E81C, one labeled with an NBD donor dye and the other with a Rh acceptor dye. Figs. 5A, 5B, Supp. Table 1 Both membranes and Vps20 are required for maximal association of Snf7 molecules. Vps20 (Q04272) and liposomes A fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye was covalently attached to the Cys mutated into the protein sequence. snf7-3 1 5 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept lys-21 rhodamine fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role fluorescence donor fluorescence donor P39929:p.Lys21Cys mutation mutation certain certain sequence position certain certain certain sequence position certain K C Wed May 02 17:31:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys21cys' Tue Jun 30 20:02:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys21Cys' cys-21 fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction The association of Snf7 into polymeric assemblies was detected directly through the use of a variation of the fluorescence resonance energy transfer (FRET) technique involving two samples of Snf7 E81C, one labeled with an NBD donor dye and the other with a Rh acceptor dye. Figs. 5A, 5B, Supp. Table 1 Both membranes and Vps20 are required for maximal association of Snf7 molecules. Vps20 (Q04272) and liposomes A fluorescent 7-nitrobenz-2-oxa-1,3-diazole (NBD) dye was covalently attached to the Cys mutated into the protein sequence. snf7-4 3 5 unspecified role unspecified role neutral component neutral component P39929:p.[Pro191Trp;Pro200Trp;Pro203Trp] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P W certain certain sequence position certain certain certain sequence position certain P W certain certain sequence position certain certain certain sequence position certain P W Fri Jul 22 10:58:51 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'pro200trp,pro203trp,pro191trp' Wed May 02 16:04:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro191trp,pro200trp,pro203trp' Tue Jun 30 20:15:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Pro191Trp;Pro200Trp;Pro203Trp]' ecoli Escherichia coli (strain K12) colocalization colocalization Fig. 5C Both membranes and Vps20 are required for maximal association of Snf7 molecules. Vps20 (Q04272) and liposomes The requirements of Vps20 and membranes for inducing Snf7 oligomerization were also observed in a nonspectroscopic assay that detected Snf7 oligomerization in vitro via velocity sedimentation centrifugation on 10%-40% glycerol gradients. Vps24 (P36095)