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/ polier-2008-2 Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Polier S., Dragovic Z., Hartl FU., Bracher A. The fluorescent probe MABA-ADP was bound to 2.5 uM of an Hsp70 protein (Ssa1p, Ssa1N or Hsp70N, as indicated) and mixed in a stopped-flow apparatus with a solution containing 250 uM ATP or ADP and either 2.5 uM nucleotide exchange factor or no protein. Release of labeled nucleotide was monitored by the decrease in fluorescence of the unbound MABA-ADP (excitation 360nm wavelength, cut-off filter 420 nm). Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de Cell (0092-8674) 2008 Only protein-protein interactions 3 imex curation polier-2008-3 Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Polier S., Dragovic Z., Hartl FU., Bracher A. Superose-12 size exclusion chromatography Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de Cell (0092-8674) 2008 Only protein-protein interactions 3 imex curation polier-2008-1 Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight by comassie molecular weight estimation by coomasie staining weight by comassie Polier S., Dragovic Z., Hartl FU., Bracher A. Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de Cell (0092-8674) 2008 Only protein-protein interactions 3 imex curation polier-2008-4 Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Polier S., Dragovic Z., Hartl FU., Bracher A. Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de Cell (0092-8674) 2008 Only protein-protein interactions 3 imex curation adp adenosine 5'-(trihydrogen diphosphate) (ADP) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) hsp71_yeast Heat shock protein SSA1 YAL005C SSA1 Heat shock protein YG100 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSKAVGIDLGTTYSCVAHFANDRVDIIANDQGNRTTPSFVAFTDTERLIGDAAKNQAAMNPSNTVFDAKRLIGRNFNDPEVQADMKHFPFKLIDVDGKPQIQVEFKGETKNFTPEQISSMVLGKMKETAESYLGAKVNDAVVTVPAYFNDSQRQATKDAGTIAGLNVLRIINEPTAAAIAYGLDKKGKEEHVLIFDLGGGTFDVSLLSIEDGIFEVKATAGDTHLGGEDFDNRLVNHFIQEFKRKNKKDLSTNQRALRRLRTACERAKRTLSSSAQTSVEIDSLFEGIDFYTSITRARFEELCADLFRSTLDPVEKVLRDAKLDKSQVDEIVLVGGSTRIPKVQKLVTDYFNGKEPNRSINPDEAVAYGAAVQAAILTGDESSKTQDLLLLDVAPLSLGIETAGGVMTKLIPRNSTIPTKKSEIFSTYADNQPGVLIQVFEGERAKTKDNNLLGKFELSGIPPAPRGVPQIEVTFDVDSNGILNVSAVEKGTGKSNKITITNDKGRLSKEDIEKMVAEAEKFKEEDEKESQRIASKNQLESIAYSLKNTISEAGDKLEQADKDTVTKKAEETISWLDSNTTASKEEFDDKLKELQDIANPIMSKLYQAGGAPGGAAGGAPGGFPGGAPPAPEAEGPTVEEVD 249D8A3BC42527CA hsp7f_yeast Heat shock protein homolog SSE1 SSE1 MSI3 YPL106C LPG3C Chaperone protein MSI3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSTPFGLDLGNNNSVLAVARNRGIDIVVNEVSNRSTPSVVGFGPKNRYLGETGKNKQTSNIKNTVANLKRIIGLDYHHPDFEQESKHFTSKLVELDDKKTGAEVRFAGEKHVFSATQLAAMFIDKVKDTVKQDTKANITDVCIAVPPWYTEEQRYNIADAARIAGLNPVRIVNDVTAAGVSYGIFKTDLPEGEEKPRIVAFVDIGHSSYTCSIMAFKKGQLKVLGTACDKHFGGRDFDLAITEHFADEFKTKYKIDIRENPKAYNRILTAAEKLKKVLSANTNAPFSVESVMNDVDVSSQLSREELEELVKPLLERVTEPVTKALAQAKLSAEEVDFVEIIGGTTRIPTLKQSISEAFGKPLSTTLNQDEAIAKGAAFICAIHSPTLRVRPFKFEDIHPYSVSYSWDKQVEDEDHMEVFPAGSSFPSTKLITLNRTGDFSMAASYTDITQLPPNTPEQIANWEITGVQLPEGQDSVPVKLKLRCDPSGLHTIEEAYTIEDIEVEEPIPLPEDAPEDAEQEFKKVTKTVKKDDLTIVAHTFGLDAKKLNELIEKENEMLAQDKLVAETEDRKNTLEEYIYTLRGKLEEEYAPFASDAEKTKLQGMLNKAEEWLYDEGFDSIKAKYIAKYEELASLGNIIRGRYLAKEEEKKQAIRSKQEASQMAAMAEKLAAQRKAEAEKKEEKKDTEGDVDMD 6558259111B0BC38 hsp71_human Heat shock 70 kDa protein 1A/1B HSPA1A HSPA1 HSPA1B Heat shock 70 kDa protein 1/2 HSX70 protein protein human Homo sapiens Human MAKAAAIGIDLGTTYSCVGVFQHGKVEIIANDQGNRTTPSYVAFTDTERLIGDAAKNQVALNPQNTVFDAKRLIGRKFGDPVVQSDMKHWPFQVINDGDKPKVQVSYKGETKAFYPEEISSMVLTKMKEIAEAYLGYPVTNAVITVPAYFNDSQRQATKDAGVIAGLNVLRIINEPTAAAIAYGLDRTGKGERNVLIFDLGGGTFDVSILTIDDGIFEVKATAGDTHLGGEDFDNRLVNHFVEEFKRKHKKDISQNKRAVRRLRTACERAKRTLSSSTQASLEIDSLFEGIDFYTSITRARFEELCSDLFRSTLEPVEKALRDAKLDKAQIHDLVLVGGSTRIPKVQKLLQDFFNGRDLNKSINPDEAVAYGAAVQAAILMGDKSENVQDLLLLDVAPLSLGLETAGGVMTALIKRNSTIPTKQTQIFTTYSDNQPGVLIQVYEGERAMTKDNNLLGRFELSGIPPAPRGVPQIEVTFDIDANGILNVTATDKSTGKANKITITNDKGRLSKEEIERMVQEAEKYKAEDEVQRERVSAKNALESYAFNMKSAVEDEGLKGKISEADKKKVLDKCQEVISWLDANTLAEKDEFEHKRKELEQVCNPIISGLYQGAGGPGPGGFGAQGPKGGSGSGPTIEEVD ssa1-sse1-2 Interaction between SSA1 and SSE1 determined by nucleotide exchange 760348 N/A 760352 unspecified role unspecified role prey prey N/A 760353 unspecified role unspecified role prey prey N/A 760354 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false P10591:p.Gln31Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P10591:p.Ala297Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 760355 stimulator stimulator prey prey P32589:p.[Asn572Tyr;Glu575Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.Ala280_Asn281delinsThrAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Thr364Val;Asn367Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.[Phe392Ala;Phe394Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.Asp396Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.Leu489_His490delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Glu554Ala;Met557_Leu558delinsSerSer] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.Leu433_Asn434delinsAlaPro mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Phe439Leu;Met441Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Figs. 4A, 4C, Supp. Figs. 1B and 8B ADP was fluorescently tagged with MABA ATP-bound Sse1p accelerated the dissociation of the fluorescent nucleotide MABA-ADP from Ssa1 and replacement with unlabelled ATP. Nucleotide exchange was less efficient when Sse1p was ADP bound. Sse1 mutations asn572tyr_glu575a and (ala280thr_asn281a or thr364val_asn367s) effectively abolishes the interaction with Ssa1. hspa1a-sse1-4 Interaction between HSPA1A/HSPA1B and SSE1 determined by nucleotide exchange 760348 N/A-x 760352 unspecified role unspecified role prey prey N/A-x 760353 unspecified role unspecified role prey prey N/A-x 760355 stimulator stimulator prey prey P32589:p.[Asn572Tyr;Glu575Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.Ala280_Asn281delinsThrAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Thr364Val;Asn367Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.[Phe392Ala;Phe394Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.Asp396Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.Leu489_His490delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Glu554Ala;Met557_Leu558delinsSerSer] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.Leu433_Asn434delinsAlaPro mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Phe439Leu;Met441Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 760356 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false Fig. 4A, Supp. 1B, Supp. Fig. 8A ADP was fluorescently tagged with MABA ATP-bound Sse1p accelerated the dissociation of the fluorescent nucleotide MABA-ADP from HSPA1A and replacement with unlabelled ATP. Sse1 mutations asn572tyr_glu575a and (ala280thr_asn281a or thr364val_asn367s) effectively abolishes the interaction with Ssa1. hspa1a-sse1-2 Interaction between HSPA1A and SSE1 determined by gel filtration 760349 N/A 760356 unspecified role unspecified role neutral component neutral component n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 760355 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Supp. Fig. 1C Complex formed in the presence of 40 uM ATP. ssa1-sse1-3 Interaction between SSA1and SSE1 determined by gel filtration 760349 N/A 760355 unspecified role unspecified role neutral component neutral component P32589:p.Ala280_Asn281delinsThrAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Phe392Ala;Phe394Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.Asp396Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.Leu489_His490delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P32589:p.[Glu554Ala;Met557Ser;Leu558Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis 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 P32589:p.[Thr365Val;Asn367Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P32589:p.[Asn572Tyr;Glu575Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 760354 unspecified role unspecified role neutral component neutral component n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P10591:p.Ala297Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P10591:p.Gln31Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Fig. 4A, 4B Complex formed in the presence of 40 uM ATP. ssa1-sse1-1 Interaction between SSA1 and SSE1 determined by pulldown 760350 N/A 760355 unspecified role unspecified role bait bait 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 ecobb Escherichia coli (strain B / BL21) N/A 760354 unspecified role unspecified role prey prey n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Supp. Fig. 1A SSE1 was pretreated with 0.2uM ATP, excessive ATP was removed with a desalting column. hspa1a-sse1-3 Interaction between HSPA1A and SSE1 determined by X-ray crystallography 760351 N/A 760353 unspecified role unspecified role neutral component neutral component N/A-x-x-x-x 760355 unspecified role unspecified role neutral component neutral component glu575_glu576 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false glu575 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false glu576 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false tyr579 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false ala279_ala280 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false asn281 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false arg346 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false lys262 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false glu304 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false undefined mutation VEEPIPLEDAPEDAEQEFKKV -> AGSD mutation mutation substitut analysis substitution analysis substitut analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A-x-x 760356 unspecified role unspecified role neutral component neutral component n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ala300 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false thr298 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false lys348 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false ala300 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false asn57 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false gln33_gly34 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false asp285 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false ala300 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false gln33 binding region binding-associated region binding region x-ray diffraction x-ray crystallography X-ray x-ray diffraction X-ray certain certain sequence position certain certain certain sequence position certain false 760426 760414 760427 760415 760428 760416 760429 760417 760430 760419 760431 760420 760432 760421 760433 760422 760434 760418 direct interaction direct interaction false false false Figs. 1,2 and 3, Supp. Table 1, Supp. Figs. 2, 3 and 4 2.35A Working 21.0%, Free 26.2%