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/ henne-2012-1 The Endosomal Sorting Complex ESCRT-II Mediates the Assembly and Architecture of ESCRT-III Helices. in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Henne W.M., Buchkovich N.J., Zhao Y., Emr S.D. sde26@cornell.edu Cell (0092-8674) 2012 Only protein-protein interactions imex curation henne-2012-2 The Endosomal Sorting Complex ESCRT-II Mediates the Assembly and Architecture of ESCRT-III Helices. yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Henne W.M., Buchkovich N.J., Zhao Y., Emr S.D. sde26@cornell.edu Cell (0092-8674) 2012 Only protein-protein interactions imex curation henne-2012-3 The Endosomal Sorting Complex ESCRT-II Mediates the Assembly and Architecture of ESCRT-III Helices. ecolx Escherichia coli pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Henne W.M., Buchkovich N.J., Zhao Y., Emr S.D. sde26@cornell.edu Cell (0092-8674) 2012 Only protein-protein interactions imex curation 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 vps24_yeast Vacuolar protein-sorting-associated protein 24 YKL041W YKL254 VPS24 DID3 DOA4-independent degradation protein 3 ESCRT-III complex subunit VPS24 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDYIKKAIWGPDPKEQQRRIRSVLRKNGRNIEKSLRELTVLQNKTQQLIKKSAKKNDVRTVRLYAKELYQINKQYDRMYTSRAQLDSVRMKIDEAIRMNTLSNQMADSAGLMREVNSLVRLPQLRNTMIELEKELMKSGIISEMVDDTMESVGDVGEEMDEAVDEEVNKIVEQYTNEKFKNVDQVPTVELAANEEEQEIPDEKVDEEADRMVNEMRERLRALQN 6073C1B680D58E98 did4_yeast DOA4-independent degradation protein 4 VPL2 VPS2 VPT14 YKL002W DID4 CHM2 GRD7 REN1 Vacuolar protein-sorting-associated protein 2 Vacuolar protein-targeting protein 14 ESCRT-III complex subunit VPS2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLFEWVFGKNVTPQERLKKNQRALERTQRELEREKRKLELQDKKLVSEIKKSAKNGQVAAAKVQAKDLVRTRNYIQKFDNMKAQLQAISLRIQAVRSSDQMTRSMSEATGLLAGMNRTMNLPQLQRISMEFEKQSDLMGQRQEFMDEAIDNVMGDEVDEDEEADEIVNKVLDEIGVDLNSQLQSTPQNLVSNAPIAETAMGIPEPIGAGSEFHGNPDDDLQARLNTLKKQT 200D07DB4C45DCBC 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 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 snf8_yeast Vacuolar-sorting protein SNF8 Vacuolar protein-sorting-associated protein 22 SNF8 ESCRT-II complex subunit VPS22 VPS22 YPL002C protein protein yeast Saccharomyces cerevisiae Baker's yeast MKQFGLAAFDELKDGKYNDVNKTILEKQSVELRDQLMVFQERLVEFAKKHNSELQASPEFRSKFMHMCSSIGIDPLSLFDRDKHLFTVNDFYYEVCLKVIEICRQTKDMNGGVISFQELEKVHFRKLNVGLDDLEKSIDMLKSLECFEIFQIRGKKFLRSVPNELTSDQTKILEICSILGYSSISLLKANLGWEAVRSKSALDEMVANGLLWIDYQGGAEALYWDPSWITRQL 212C74F086A0325E vps36_yeast Vacuolar protein-sorting-associated protein 36 VPS36 GRD12 VAC3 VPL11 YLR417W ESCRT-II complex subunit VPS36 protein protein yeast Saccharomyces cerevisiae Baker's yeast MEYWHYVETTSSGQPLLREGEKDIFIDQSVGLYHGKSKILQRQRGRIFLTSQRIIYIDDAKPTQNSLGLELDDLAYVNYSSGFLTRSPRLILFFKDPSSKDELGKSAETASADVVSTWVCPICMVSNETQGEFTKDTLPTPICINCGVPADYELTKSSINCSNAIDPNANPQNQFGVNSENICPACTFANHPQIGNCEICGHRLPNASKVRSKLNRLNFHDSRVHIELEKNSLARNKSSHSALSSSSSTGSSTEFVQLSFRKSDGVLFSQATERALENILTEKNKHIFNQNVVSVNGVDMRKGASSHEYNNEVPFIETKLSRIGISSLEKSRENQLLNNDILFNNALTDLNKLMSLATSIERLYKNSNITMKTKTLNLQDESTVNEPKTRRPLLILDREKFLNKELFLDEIAREIYEFTLSEFKDLNSDTNYMIITLVDLYAMYNKSMRIGTGLISPMEMREACERFEHLGLNELKLVKVNKRILCVTSEKFDVVKEKLVDLIGDNPGSDLLRLTQILSSNNSKSNWTLGILMEVLQNCVDEGDLLIDKQLSGIYYYKNSYWPSHI 60DDD4EA3A620409 snf7-1 1331426 n/a 1331429 unspecified role unspecified role neutral component neutral component P39929:p.Arg52Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false P39929:p.[Glu160Lys;Glu162Lys;Glu165_Glu166delinsLysLys] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1331429 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false P39929:p.Arg52Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false P39929:p.[Glu160Lys;Glu162Lys;Glu165_Glu166delinsLysLys] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1331444 1331441 direct interaction direct interaction false false false Fig. 3, 4A-B, 4D-F, 5C-D, 6B, Supp. Fig. 3 Complex formed protofilaments did4-snf7 1331426 n/a 1331429 unspecified role unspecified role neutral component neutral component P39929:p.Arg52Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 n/a 1331430 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 n/a 1331431 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 physical association physical association false false false Fig. 5E-I Molecules form a polymeric structure vps36-snf8-2 1331426 n/a 1331432 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331429 unspecified role unspecified role neutral component neutral component P39929:p.Ile53Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli n/a 1331433 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331434 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331435 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli physical association physical association false false false Fig. 6C, 6E, Supp. Fig. 7A, 7D Complex formed rings composed of highly curved ∼9 nm diameter protofilaments . Ring formation required the presence of ESCRT-II, Vps20, and Snf7R52E, as removal of either ESCRT-II or Vps20 drastically reduced ring formation in favor of linear protofilaments snf7-snf8 1331426 n/a 1331435 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli n/a 1331434 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331432 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331429 unspecified role unspecified role neutral component neutral component P39929:p.Ile53Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli physical association physical association false false false Supp. Fig. 7B Complex formed protofilaments snf7-snf8-1 1331426 n/a 1331433 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331429 unspecified role unspecified role neutral component neutral component P39929:p.Ile53Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli physical association physical association false false false Supp. Fig. 7C Complex formed rings composed of highly curved ∼9 nm diameter protofilaments . Ring formation required the presence of ESCRT-II, Vps20, and Snf7R52E, as removal of either ESCRT-II or Vps20 drastically reduced ring formation in favor of linear protofilaments did4-snf7-3 1331426 n/a 1331431 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331435 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecolx Escherichia coli n/a 1331433 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331434 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331432 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli n/a 1331429 unspecified role unspecified role neutral component neutral component P39929:p.Ile53Glu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecolx Escherichia coli n/a 1331430 unspecified role unspecified role neutral component neutral component ecolx Escherichia coli physical association physical association false false false Supp. Fig. 7E Complex formed rings composed of highly curved ∼9 nm diameter protofilaments . snf7-snf7-1 1331427 n/a 1331429 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false n/a 1331429 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig.4C-F Tagged and untagged proteins differentiated by weight. vps36-snf8-1 1331428 n/a 1331434 unspecified role unspecified role prey prey n/a 1331435 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1331432 unspecified role unspecified role prey prey association association false false false Fig. 6A