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/ Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Cell (0092-8674) 2008 imex curation Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library davet Only protein-protein interactions 9 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk Accepted 2008-NOV-2 by DAVET. 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Cell (0092-8674) 2008 imex curation Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library davet Only protein-protein interactions 9 in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk Accepted 2008-NOV-2 by DAVET. 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Cell (0092-8674) 2008 imex curation Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library davet Only protein-protein interactions 9 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk Accepted 2008-NOV-2 by DAVET. 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Cell (0092-8674) 2008 imex curation Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library davet Only protein-protein interactions 9 in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. Authors contacted - no reply to date Complex formed with SNARE and STX-4 could not be added as source of protein could not be determined. djo30@cam.ac.uk, jpl10@cam.ac.uk Accepted 2008-NOV-2 by DAVET. 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Cell (0092-8674) 2008 imex curation Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library davet Only protein-protein interactions 9 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk Accepted 2008-NOV-2 by DAVET. 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Molecular basis for the sorting of the SNARE VAMP7 into endocytic clathrin-coated vesicles by the ArfGAP Hrb. Cell (0092-8674) 2008 imex curation Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library davet Only protein-protein interactions 9 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Pryor PR., Jackson L., Gray SR., Edeling MA., Thompson A., Sanderson CM., Evans PR., Owen DJ., Luzio JP. djo30@cam.ac.uk, jpl10@cam.ac.uk Accepted 2008-NOV-2 by DAVET. 16-Oct-2008: Contacted by JYOTI. K562 erythroleukemia cDNA Matchmaker library Cell (0092-8674) 2008 davet Only protein-protein interactions 9 imex curation vamp7_hrb fusion Fusion of mouse Vamp7 (1-120) and human Hrb (136-176) protein protein human Homo sapiens Human VASVHASISGSSASSTSSTPEVKPLKSLLGDSAPTLHLNKMAILFAVVARGTTILAKHAWCGGNFLEVTEQILAKIPSENNKLTYSHGNYLFHYICQDRIVYLCITDDDFERSRAFSFLNEVKKRFQTTYGSRAQTALPYAMNSEFSSVLAAQLKHHSEN An amino acid sequence which is not represented in UniProt 75C175257F670971 ap3d1_human AP-3 complex subunit delta-1 AP-3 complex subunit delta Delta-adaptin AP3D1 PRO0039 Adaptor-related protein complex 3 subunit delta-1 protein protein human Homo sapiens Human MALKMVKGSIDRMFDKNLQDLVRGIRNHKEDEAKYISQCIDEIKQELKQDNIAVKANAVCKLTYLQMLGYDISWAAFNIIEVMSASKFTFKRIGYLAASQSFHEGTDVIMLTTNQIRKDLSSPSQYDTGVALTGLSCFVTPDLARDLANDIMTLMSHTKPYIRKKAVLIMYKVFLKYPESLRPAFPRLKEKLEDPDPGVQSAAVNVICELARRNPKNYLSLAPLFFKLMTSSTNNWVLIKIIKLFGALTPLEPRLGKKLIEPLTNLIHSTSAMSLLYECVNTVIAVLISLSSGMPNHSASIQLCVQKLRILIEDSDQNLKYLGLLAMSKILKTHPKSVQSHKDLILQCLDDKDESIRLRALDLLYGMVSKKNLMEIVKKLMTHVDKAEGTTYRDELLTKIIDICSQSNYQYITNFEWYISILVELTRLEGTRHGHLIAAQMLDVAIRVKAIRKFAVSQMSALLDSAHLLASSTQRNGICEVLYAAAWICGEFSEHLQEPHHTLEAMLRPRVTTLPGHIQAVYVQNVVKLYASILQQKEQAGEAEGAQAVTQLMVDRLPQFVQSADLEVQERASCILQLVKHIQKLQAKDVPVAEEVSALFAGELNPVAPKAQKKVPVPEGLDLDAWINEPLSDSESEDERPRAVFHEEEQRRPKHRPSEADEEELARRREARKQEQANNPFYIKSSPSPQKRYQDTPGVEHIPVVQIDLSVPLKVPGLPMSDQYVKLEEERRHRQKLEKDKRRKKRKEKEKKGKRRHSSLPTESDEDIAPAQQVDIVTEEMPENALPSDEDDKDPNDPYRALDIDLDKPLADSEKLPIQKHRNTETSKSPEKDVPMVEKKSKKPKKKEKKHKEKERDKEKKKEKEKKKSPKPKKKKHRKEKEERTKGKKKSKKQPPGSEEAAGEPVQNGAPEEEQLPPESSYSLLAENSYVKMTCDIRGSLQEDSQVTVAIVLENRSSSILKGMELSVLDSLNARMARPQGSSVHDGVPVPFQLPPGVSNEAQYVFTIQSIVMAQKLKGTLSFIAKNDEGATHEKLDFRLHFSCSSYLITTPCYSDAFAKLLESGDLSMSSIKVDGIRMSFQNLLAKICFHHHFSVVERVDSCASMYSRSIQGHHVCLLVKKGENSVSVDGKCSDSTLLSNLLEEMKATLAKC DIP protein O14617 original sequence version: 119 1B491DD64EAD5096 vamp7_human Vesicle-associated membrane protein 7 SYBL1 Synaptobrevin-like protein 1 Tetanus-insensitive VAMP VAMP7 protein protein human Homo sapiens Human MAILFAVVARGTTILAKHAWCGGNFLEVTEQILAKIPSENNKLTYSHGNYLFHYICQDRIVYLCITDDDFERSRAFNFLNEIKKRFQTTYGSRAQTALPYAMNSEFSSVLAAQLKHHSENKGLDKVMETQAQVDELKGIMVRNIDLVAQRGERLELLIDKTENLVDSSVTFKTTSRNLARAMCMKNLKLTIIIIIVSIVFIYIIVSPLCGGFTWPSCVKK 9C1AA5C590375CEF agfg1_human Arf-GAP domain and FG repeat-containing protein 1 HIV-1 Rev-binding protein Rev-interacting protein Rev/Rex activation domain-binding protein AGFG1 RAB RIP HRB Nucleoporin-like protein RIP protein protein human Homo sapiens Human MAASAKRKQEEKHLKMLRDMTGLPHNRKCFDCDQRGPTYVNMTVGSFVCTSCSGSLRGLNPPHRVKSISMTTFTQQEIEFLQKHGNEVCKQIWLGLFDDRSSAIPDFRDPQKVKEFLQEKYEKKRWYVPPEQAKVVASVHASISGSSASSTSSTPEVKPLKSLLGDSAPTLHLNKGTPSQSPVVGRSQGQQQEKKQFDLLSDLGSDIFAAPAPQSTATANFANFAHFNSHAAQNSANADFANFDAFGQSSGSSNFGGFPTASHSPFQPQTTGGSAASVNANFAHFDNFPKSSSADFGTFNTSQSHQTASAVSKVSTNKAGLQTADKYAALANLDNIFSAGQGGDQGSGFGTTGKAPVGSVVSVPSQSSASSDKYAALAELDSVFSSAATSSNAYTSTSNASSNVFGTVPVVASAQTQPASSSVPAPFGATPSTNPFVAAAGPSVASSTNPFQTNARGATAATFGTASMSMPTGFGTPAPYSLPTSFSGSFQQPAFPAQAAFPQQTAFSQQPNGAGFAAFGQTKPVVTPFGQVAAAGVSSNPFMTGAPTGQFPTGSSSTNPFL DIP protein P52594 original sequence version: 121 9A2242217F53B4D9 vamp7-hrb-1 6 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association vamp7-ap3d1-1 6 9 unspecified role unspecified role bait bait longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey physical association physical association To identify proteins that bind the longin domain of VAMP7, a yeast two-hybrid cDNA library screen using the longin domain as the bait protein was carried out. In this screen, 37 out of 39 positive clones encoded part of the AP3 complex δ subunit trunk domain. vamp7-hrb-2 1 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association To test the specificity of the interaction between the longin domain of VAMP7 and Hrb, we first used the yeast two-hybrid system t Fig. 1B vamp7-hrb-3 4 10 unspecified role unspecified role prey prey c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region 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) 9 unspecified role unspecified role bait bait longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 28 26 direct interaction direct interaction The binding of various C-terminally His6-tagged Hrb constructs to GST-VAMP7 longin domain demonstrates that the Hrb binding site for VAMP7 longin domain is located between residues 136 and 176. Figs. 1D, 1E, 7B, 7C The full-length, wild-type cytoplasmic domain of VAMP7 does not bind HRB due to the intramolecular binding of the VAMP7 SNARE helix onto its longin domain. Mutations (Ile139, Met140, Ile144 -> Ser) prevents intramolecular binding and allows HRB binding. vamp7-hrb-4 5 10 unspecified role unspecified role prey prey human-a_431 Homo sapiens epidermoid carcinoma a_431 Human epidermoid carcinoma 9 unspecified role unspecified role bait bait longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) physical association physical association GST pull-down experiments indicate that VAMP7 longin domain but not Sec22b longin domain or GST alone binds full-length Hrb from A431 cell cytosol shown by immunoblotting with an anti-Hrb antibody. Figs. 1C vamp7-hrb-5 2 10 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P52594:p.[Leu160Ser;Leu163Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S certain certain sequence position certain certain certain sequence position certain L S Fri Jul 22 11:11:51 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu163ser,leu160ser' Wed May 02 14:46:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu160ser,leu163ser' Tue Jun 30 20:03:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu160Ser;Leu163Ser]' P52594:p.[Leu163Ser;Leu164Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S certain certain sequence position certain certain certain sequence position certain L S Wed May 02 16:25:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu163ser,leu164ser' Tue Jun 30 19:23:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu163Ser;Leu164Ser]' P52594:p.[Leu171Ser;Leu173Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S certain certain sequence position certain certain certain sequence position certain L S Wed May 02 19:01:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu171ser,leu173ser' Tue Jun 30 18:01:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu171Ser;Leu173Ser]' ecobb Escherichia coli (strain B / BL21) 9 unspecified role unspecified role neutral component neutral component longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P51809:p.[Leu43Ser;Tyr45Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain Y S certain certain sequence position certain certain certain sequence position certain L S Fri Jul 22 10:54:43 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr45ser,leu43ser' Wed May 02 19:32:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu43ser,tyr45ser' Tue Jun 30 17:59:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu43Ser;Tyr45Ser]' ecobb Escherichia coli (strain B / BL21) 43 38 direct interaction direct interaction Using residues 1�176 of Hrb and the VAMP7 longin domain in isothermal titration calorimetry (ITC), the KD for the interaction was measured as 10.5 μM Figs. 4C All mutation increased the Kd to >3E-4M. vamp7-hrb-6 3 7 unspecified role unspecified role self self c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range direct interaction direct interaction true working 22.0%, free 28.0% 2.2A Structure Determination of VAMP7 Longin Domain/Hrb136-176 Complex Fig. 3, Supp. Table 1 vamp7-hrb-7 4 10 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 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) 9 unspecified role unspecified role prey prey longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 P51809:p.[Leu43Ser;Tyr45Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S certain certain sequence position certain certain certain sequence position certain Y S Wed May 02 16:30:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu43ser,tyr45ser' Tue Jun 30 17:55:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu43Ser;Tyr45Ser]' ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Interaction of GST Hrb(1-176) and GST control proteins with Wt and mutant versions of VAMP7 longin domain His6 as indicated by western blotting of 'GST pull down' experiments with anti His6 antibody. Supp. Fig. 2D vamp7-hrb-8 4 10 unspecified role unspecified role prey prey c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain P52594:p.[Leu160Ser;Leu163Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S certain certain sequence position certain certain certain sequence position certain L S Fri Jul 22 11:12:13 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu163ser,leu160ser' Wed May 02 15:31:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu160ser,leu163ser' Tue Jun 30 17:05:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu160Ser;Leu163Ser]' P52594:p.[Leu163Ser;Leu164Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S certain certain sequence position certain certain certain sequence position certain L S Fri Jul 22 11:14:06 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'leu164ser,leu163ser' Wed May 02 18:11:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu163ser,leu164ser' Tue Jun 30 16:36:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu163Ser;Leu164Ser]' P52594:p.[Leu171Ser;Leu173Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L S certain certain sequence position certain certain certain sequence position certain L S Wed May 02 15:35:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu171ser,leu173ser' Tue Jun 30 19:58:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu171Ser;Leu173Ser]' ecobb Escherichia coli (strain B / BL21) 9 unspecified role unspecified role bait bait longin domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Interaction of GST VAMP7 longin domain and GST control proteins with Wt and mutant versions of Hrb(1-176)His6 as indicated by western blotting of 'GST pull down' experiments with anti His antibody. Supp. Fig. 2E