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 basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Cell (0092-8674) 2008 imex curation Polier S., Dragovic Z., Hartl FU., Bracher A. Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 3 in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Polier S., Dragovic Z., Hartl FU., Bracher A. Data from figs 5 and Supp Figs 9-11 was not captured as this proved too complex for the data model 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 Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 3 imex curation Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Cell (0092-8674) 2008 imex curation Polier S., Dragovic Z., Hartl FU., Bracher A. Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 3 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 Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 3 imex curation Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Cell (0092-8674) 2008 imex curation Polier S., Dragovic Z., Hartl FU., Bracher A. Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 3 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 Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 3 imex curation Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Structural basis for the cooperation of Hsp70 and Hsp110 chaperones in protein folding. Cell (0092-8674) 2008 imex curation Polier S., Dragovic Z., Hartl FU., Bracher A. Additional methodology supplied by the authors. uhartl@biochem.mpg.de; bracher@biochem.mpg.de 16-Oct-2008: Contacted by JYOTI. davet Only protein-protein interactions 3 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 Accepted 2008-OCT-29 by DAVET. 16-Oct-2008: Contacted by JYOTI. Cell (0092-8674) 2008 davet Only protein-protein interactions 3 imex curation 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 78F513118C96DE66 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) adp adenosine 5'-(trihydrogen diphosphate) (ADP) 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 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 6558259111B0BC38 ssa1-sse1-1 4 9 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 ecobb Escherichia coli (strain B / BL21) 8 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction The KD value for complex formation between Sse1p and the NBD of Ssa1p, Ssa1N, was estimated from the distribution of Ssa1N between supernatant and Ni2+ resin-bound His-tagged Sse1p at different concentrations. Supp. Fig. 1A SSE1 was pretreated with 0.2uM ATP, excessive ATP was removed with a desalting column. ssa1-sse1-2 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 8 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 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 Q A Wed May 02 20:02:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln31ala' Tue Jun 30 17:03:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln31Ala' 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 A E Wed May 02 15:50:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala297glu' Tue Jun 30 19:02:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala297Glu' ecobb Escherichia coli (strain B / BL21) 9 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 E A certain certain sequence position certain certain certain sequence position certain N Y Fri Jul 22 10:53:06 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu575ala,asn572tyr' Wed May 02 18:56:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn572tyr,glu575ala' Tue Jun 30 20:09:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn572Tyr;Glu575Ala]' 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 AN TA Thu Jul 07 12:16:50 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ala280thr_asn281a Fri Jul 22 10:55:56 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ala280thr,asn281ala' Wed May 02 15:07:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala_asn280-281thr_ala' Tue Jun 30 18:13:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala280_Asn281delinsThrAla' 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 N S certain certain sequence position certain certain certain sequence position certain T V Fri Jul 22 11:11:32 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'asn367ser,thr364val' Wed May 02 20:07:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr364val,asn367ser' Tue Jun 30 17:38:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Thr364Val;Asn367Ser]' 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 F A certain certain sequence position certain certain certain sequence position certain F A Wed May 02 19:52:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe392ala,phe394ala' Tue Jun 30 19:59:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe392Ala;Phe394Ala]' 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 D A Wed May 02 19:39:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp396ala' Tue Jun 30 16:36:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp396Ala' 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 LH AA Thu Jul 07 12:16:19 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu489ala_his490a Fri Jul 22 11:01:10 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu489ala,his490ala' Wed May 02 14:52:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_his489-490ala_ala' Tue Jun 30 18:22:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu489_His490delinsAlaAla' 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 E A certain certain sequence position certain certain certain sequence position certain ML SS Thu Jul 07 12:16:19 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was e554am557sl558s Fri Jul 22 10:58:49 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu554ala,met557ser,leu558ser' Wed May 02 14:58:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu554ala,met_leu557-558ser_ser' Tue Jun 30 16:33:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu554Ala;Met557_Leu558delinsSerSer]' 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 LN AP Thu Jul 07 12:16:13 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu433ala_asn434p Fri Jul 22 10:52:11 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu433ala,asn434pro' Wed May 02 18:23:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_asn433-434ala_pro' Tue Jun 30 18:05:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu433_Asn434delinsAlaPro' 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 M A certain certain sequence position certain certain certain sequence position certain F L Fri Jul 22 11:05:28 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'met441ala,phe439leu' Wed May 02 16:52:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe439leu,met441ala' Tue Jun 30 19:03:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe439Leu;Met441Ala]' ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction ATP-bound Sse1p accelerated the dissociation of the fluorescent nucleotide MABA-ADP from Ssa1N or full-length Ssa1p to a similar extent, indicating that the interaction of cytosolic Hsp110s with the NBD of Hsp70 is sufficient for binding and nucleotide exchange (Figure S1B). 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-sse1-2 2 5 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 9 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Sse1p or Sse1-Δloop were mixed with Hsp70N (17.2 μM each) in presence of 40 μM ATP and applied to a Superose-12 size exclusion chromatography column Supp. Fig. 1C Complex formed in the presence of 40 uM ATP. hspa1a-sse1-3 3 6 unspecified role unspecified role neutral component neutral component 9 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 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 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 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 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 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 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 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 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 undefined mutation VEEPIPLEDAPEDAEQEFKKV -> AGSD mutation mutation substitut analysis substitution analysis substitut analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:25:43 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'agsd loop region' ecobb Escherichia coli (strain B / BL21) 5 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 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 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 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 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 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 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 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 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 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 39 51 52 40 41 53 54 42 59 43 44 55 45 56 46 57 58 47 direct interaction direct interaction Crystals were obtained for a complex of Hsp70N and a truncated variant of Sse1p, Sse1-Δloop, in which the poorly conserved and presumably flexible insertion (residues 503�524) found in all Hsp110/Hsp170 homologs is replaced by the peptide linker sequence AGSD. Figs. 1,2 and 3, Supp. Table 1, Supp. Figs. 2, 3 and 4 2.35A Working 21.0%, Free 26.2% ssa1-sse1-3 2 9 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 AN TA Thu Jul 07 12:15:45 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ala280thr_asn281a Fri Jul 22 10:56:39 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ala280thr,asn281ala' Wed May 02 15:03:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala_asn280-281thr_ala' Tue Jun 30 20:21:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala280_Asn281delinsThrAla' 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 F A certain certain sequence position certain certain certain sequence position certain F A Wed May 02 16:37:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe392ala,phe394ala' Tue Jun 30 17:13:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe392Ala;Phe394Ala]' 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 D A Wed May 02 16:27:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp396ala' Tue Jun 30 18:33:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp396Ala' 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 LH AA Thu Jul 07 12:15:55 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu489ala_his490a Fri Jul 22 10:54:23 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu489ala,his490ala' Wed May 02 18:17:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_his489-490ala_ala' Tue Jun 30 19:47:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu489_His490delinsAlaAla' 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 E A certain certain sequence position certain certain certain sequence position certain M S certain certain sequence position certain certain certain sequence position certain L S Wed May 02 19:50:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu554ala,met557ser,leu558ser' Tue Jun 30 18:25:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu554Ala;Met557Ser;Leu558Ser]' 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 N S certain certain sequence position certain certain certain sequence position certain T V Fri Jul 22 10:54:21 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'asn367ser,thr365val' Wed May 02 14:52:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr365val,asn367ser' Tue Jun 30 17:16:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Thr365Val;Asn367Ser]' 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 N Y certain certain sequence position certain certain certain sequence position certain E A Wed May 02 17:45:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn572tyr,glu575ala' Tue Jun 30 16:29:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn572Tyr;Glu575Ala]' ecobb Escherichia coli (strain B / BL21) 8 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 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 A E Wed May 02 14:27:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala297glu' Tue Jun 30 17:54:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala297Glu' 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 Q A Wed May 02 19:20:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln31ala' Tue Jun 30 19:05:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln31Ala' ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Interaction of Sse1p mutants with Ssa1N analyzed by size exclusion chromatography. Fig. 4A, 4B Complex formed in the presence of 40 uM ATP. hspa1a-sse1-4 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 9 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 E A certain certain sequence position certain certain certain sequence position certain N Y Fri Jul 22 11:00:15 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu575ala,asn572tyr' Wed May 02 14:34:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn572tyr,glu575ala' Tue Jun 30 20:09:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn572Tyr;Glu575Ala]' 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 AN TA Thu Jul 07 12:15:46 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ala280thr_asn281a Fri Jul 22 10:53:21 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ala280thr,asn281ala' Wed May 02 16:51:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala_asn280-281thr_ala' Tue Jun 30 20:23:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala280_Asn281delinsThrAla' 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 N S certain certain sequence position certain certain certain sequence position certain T V Fri Jul 22 10:59:09 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'asn367ser,thr364val' Wed May 02 17:31:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr364val,asn367ser' Tue Jun 30 16:45:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Thr364Val;Asn367Ser]' 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 F A certain certain sequence position certain certain certain sequence position certain F A Wed May 02 16:43:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe392ala,phe394ala' Tue Jun 30 18:33:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe392Ala;Phe394Ala]' 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 D A Wed May 02 18:31:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp396ala' Tue Jun 30 18:44:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp396Ala' 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 LH AA Thu Jul 07 12:15:54 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu489ala_his490a Fri Jul 22 11:01:14 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu489ala,his490ala' Wed May 02 16:44:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_his489-490ala_ala' Tue Jun 30 17:20:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu489_His490delinsAlaAla' 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 ML SS certain certain sequence position certain certain certain sequence position certain E A Thu Jul 07 12:15:55 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was e554am557sl558s Fri Jul 22 11:12:21 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'glu554ala,met557ser,leu558ser' Wed May 02 19:54:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu554ala,met_leu557-558ser_ser' Tue Jun 30 15:55:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu554Ala;Met557_Leu558delinsSerSer]' 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 LN AP Thu Jul 07 12:15:53 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was leu433ala_asn434p Fri Jul 22 11:01:40 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu433ala,asn434pro' Wed May 02 19:34:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_asn433-434ala_pro' Tue Jun 30 18:59:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu433_Asn434delinsAlaPro' 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 M A certain certain sequence position certain certain certain sequence position certain F L Fri Jul 22 11:12:43 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'met441ala,phe439leu' Wed May 02 19:12:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe439leu,met441ala' Tue Jun 30 17:50:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe439Leu;Met441Ala]' ecobb Escherichia coli (strain B / BL21) 5 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 direct interaction direct interaction The fluorescent probe MABA-ADP was bound to 2.5 μM Hsp70N, and mixed in a stopped-flow apparatus with a solution containing 250 μM ATP and 2.5 μM Sse1p variant or no protein (spontaneous) 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.