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/ TSG-6 inhibits neutrophil migration via direct interaction with the chemokine CXCL8. TSG-6 inhibits neutrophil migration via direct interaction with the chemokine CXCL8. Journal of immunology (Baltimore, Md. : 1950) 2014 imex curation Dyer DP., Thomson JM., Hermant A., Jowitt TA., Handel TM., Proudfoot AE., Day AJ., Milner CM. I have not entered CXCL8/IL8 as PRO_0000005128 as although mapping would indicate this form has been used, it is not stated in Ref 32 and I would not assume that the mutation numbering is correctly stated (other statements are not correct) so I have entered mutations in relation to the the full length sequence for safety. (Sandra agreed this is OK) Dark Space = 29? CCL3, CCL5, and CXCL11 (provided by Amanda E. I. Proudfoot) have been entered with a Human origin, based on previous work of Ms Proudfoot: see pmids:25907556, 25266725, 20363748 ( from PDB entry). TSG-6 Link region has been entered as amino acids 36-133 -as quoted in both references 27 and 29. However reference 31 gives different residues. CXCL8/IL8 (also obtained from Ms Proudfoot/Pharmaceutical Research Laboratory, Geneva)- residue numbering has been entered here with respect to the full length sequence, including signal sequence. From mutation mapping IL-8(6-77) - the most predominant form, may have been used, but this is not stated in Ref 32. caroline.milner@manchester.ac.uk,anthony.day@manchester.ac.uk 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Dyer DP., Thomson JM., Hermant A., Jowitt TA., Handel TM., Proudfoot AE., Day AJ., Milner CM. I have not entered CXCL8/IL8 as PRO_0000005128 as although mapping would indicate this form has been used, it is not stated in Ref 32 and I would not assume that the mutation numbering is correctly stated (other statements are not correct) so I have entered mutations in relation to the the full length sequence for safety. (Sandra agreed this is OK) CCL3, CCL5, and CXCL11 (provided by Amanda E. I. Proudfoot) have been entered with a Human origin, based on previous work of Ms Proudfoot: see pmids:25907556, 25266725, 20363748 ( from PDB entry). TSG-6 Link region has been entered as amino acids 36-133 -as quoted in both references 27 and 29. However reference 31 gives different residues. CXCL8/IL8 (also obtained from Ms Proudfoot/Pharmaceutical Research Laboratory, Geneva)- residue numbering has been entered here with respect to the full length sequence, including signal sequence. From mutation mapping IL-8(6-77) - the most predominant form, may have been used, but this is not stated in Ref 32. caroline.milner@manchester.ac.uk,anthony.day@manchester.ac.uk Accepted 2016-JUN-02 AT 10:16 BST AT 10:16 BST by ORCHARD Journal of immunology (Baltimore, Md. : 1950) 2014 Only protein-protein interactions imex curation imex curation altered.; all altered, I think TSG-6 inhibits neutrophil migration via direct interaction with the chemokine CXCL8. TSG-6 inhibits neutrophil migration via direct interaction with the chemokine CXCL8. Journal of immunology (Baltimore, Md. : 1950) 2014 imex curation Dyer DP., Thomson JM., Hermant A., Jowitt TA., Handel TM., Proudfoot AE., Day AJ., Milner CM. I have not entered CXCL8/IL8 as PRO_0000005128 as although mapping would indicate this form has been used, it is not stated in Ref 32 and I would not assume that the mutation numbering is correctly stated (other statements are not correct) so I have entered mutations in relation to the the full length sequence for safety. (Sandra agreed this is OK) Dark Space = 29? CCL3, CCL5, and CXCL11 (provided by Amanda E. I. Proudfoot) have been entered with a Human origin, based on previous work of Ms Proudfoot: see pmids:25907556, 25266725, 20363748 ( from PDB entry). TSG-6 Link region has been entered as amino acids 36-133 -as quoted in both references 27 and 29. However reference 31 gives different residues. CXCL8/IL8 (also obtained from Ms Proudfoot/Pharmaceutical Research Laboratory, Geneva)- residue numbering has been entered here with respect to the full length sequence, including signal sequence. From mutation mapping IL-8(6-77) - the most predominant form, may have been used, but this is not stated in Ref 32. caroline.milner@manchester.ac.uk,anthony.day@manchester.ac.uk 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined pH 6 7.2 Dyer DP., Thomson JM., Hermant A., Jowitt TA., Handel TM., Proudfoot AE., Day AJ., Milner CM. Table 1 gives a value of 15,100 for the Kd for the rhTSG-6/CCL3 interaction;this is a negative result and not entered. Kds:-I have added each SPR figure separately rather than attempting to add Kds to mutation features- so i can see what I am doing....and i dont see how you can add the varying pH otherwise. I have not entered CXCL8/IL8 as PRO_0000005128 as although mapping would indicate this form has been used, it is not stated in Ref 32 and I would not assume that the mutation numbering is correctly stated (other statements are not correct) so I have entered mutations in relation to the the full length sequence for safety. (Sandra agreed this is OK) CCL3, CCL5, and CXCL11 (provided by Amanda E. I. Proudfoot) have been entered with a Human origin, based on previous work of Ms Proudfoot: see pmids:25907556, 25266725, 20363748 ( from PDB entry). TSG-6 Link region has been entered as amino acids 36-133 -as quoted in both references 27 and 29. However reference 31 gives different residues. CXCL8/IL8 (also obtained from Ms Proudfoot/Pharmaceutical Research Laboratory, Geneva)- residue numbering has been entered here with respect to the full length sequence, including signal sequence. From mutation mapping IL-8(6-77) - the most predominant form, may have been used, but this is not stated in Ref 32. caroline.milner@manchester.ac.uk,anthony.day@manchester.ac.uk Accepted 2016-JUN-02 AT 10:16 BST AT 10:16 BST by ORCHARD Journal of immunology (Baltimore, Md. : 1950) 2014 Only protein-protein interactions imex curation imex curation altered.; all altered, I think ccl5_human C-C motif chemokine 5 CCL5 D17S136E SCYA5 EoCP Eosinophil chemotactic cytokine SIS-delta Small-inducible cytokine A5 T cell-specific protein P228 T-cell-specific protein RANTES protein protein human Homo sapiens Human MKVSAAALAVILIATALCAPASASPYSSDTTPCCFAYIARPLPRAHIKEYFYTSGKCSNPAVVFVTRKNRQVCANPEKKWVREYINSLEMS DIP protein P13501 original sequence version: 123 DIP protein P13501 original sequence version: 133 DIP protein P13501 original sequence version: 170 FB0BFAF9A87C620F tsg6_human Tumor necrosis factor-inducible gene 6 protein TSG6 TNFAIP6 Hyaluronate-binding protein TNF-stimulated gene 6 protein Tumor necrosis factor alpha-induced protein 6 protein protein human Homo sapiens Human MIILIYLFLLLWEDTQGWGFKDGIFHNSIWLERAAGVYHREARSGKYKLTYAEAKAVCEFEGGHLATYKQLEAARKIGFHVCAAGWMAKGRVGYPIVKPGPNCGFGKTGIIDYGIRLNRSERWDAYCYNPHAKECGGVFTDPKQIFKSPGFPNEYEDNQICYWHIRLKYGQRIHLSFLDFDLEDDPGCLADYVEIYDSYDDVHGFVGRYCGDELPDDIISTGNVMTLKFLSDASVTAGGFQIKYVAMDPVSKSSQGKNTSTTSTGNKNFLAGRFSHL E2B3AEB76353A782 heparin Heparin small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) il8_human Interleukin-8 Protein 3-10C T-cell chemotactic factor CXCL8 C-X-C motif chemokine 8 Chemokine (C-X-C motif) ligand 8 Emoctakin Granulocyte chemotactic protein 1 IL8 Monocyte-derived neutrophil chemotactic factor Monocyte-derived neutrophil-activating peptide Neutrophil-activating protein 1 protein protein human Homo sapiens Human MTSKLAVALLAAFLISAALCEGAVLPRSAKELRCQCIKTYSKPFHPKFIKELRVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS DIP protein P10145 original sequence version: 161 15C649996E89319F cxl11_human C-X-C motif chemokine 11 CXCL11 ITAC SCYB11 SCYB9B Beta-R1 H174 Interferon gamma-inducible protein 9 Interferon-inducible T-cell alpha chemoattractant Small-inducible cytokine B11 protein protein human Homo sapiens Human MSVKGMAIALAVILCATVVQGFPMFKRGRCLCIGPGVKAVKVADIEKASIMYPSNNCDKIEVIITLKENKGQRCLNPKSKQARLIIKKVERKNF 9D9D8585749EA4CE tnfaip6-cxcl8-2 1 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 2A tnfaip6-cxcl8-1 1 6 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 2B tnfaip6-cxcl8-3 1 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 2Ai cxcl8-tnfaip6-1 1 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction 2B-1 cxcl8-tnfaip6-2 1 7 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait P10145:p.Arg95Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A Wed May 02 16:51:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg95ala' Tue Jun 30 17:15:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg95Ala' P10145:p.[Lys91Ala;Lys94_Arg95delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain KR AA certain certain sequence position certain certain certain sequence position certain K A Fri Jul 22 10:58:29 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys91ala,lys94ala,arg95ala' Wed May 02 15:34:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys91ala,lys_arg94-95ala_ala' Tue Jun 30 18:36:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys91Ala;Lys94_Arg95delinsAlaAla]' direct interaction direct interaction 3A It is stated:-"KD values of 5 ± 12 and 26 ± 11 nM were determined for the binding of b-heparin to WT CXCL8 and CXCL8_D, respectively (A)." However no construct with the name of CXCL8_D is described elsewhere. cxcl8-heparin-1 1 7 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 competitor competitor competitor prey prey P98066:p.[Lys69Ala;Lys89Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:52:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys69ala,lys89ala' Tue Jun 30 19:04:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys69Ala;Lys89Ala]' P98066:p.[Lys55Ala;Lys69Ala;Lys76Ala] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 16:34:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys55ala,lys69ala,lys76ala' Tue Jun 30 19:55:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys55Ala;Lys69Ala;Lys76Ala]' link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role bait bait direct interaction direct interaction 3B-1/C I have entered the ic50 as 70nM for WT Link_TSG6 (at interaction level) - however the value of 105 is also given.- i may have missed something here, as I cannot see what the difference is between these two assays - i dont see any... Also from text under fig 3:- "whereas Link_TSG6_T showed enhanced activity with maximal inhibition at 0.125 μM (Fig. 3C)" - so this would translate to an IC50 of ~60nM? -similar to "70". cxcl8-heparin-2 1 7 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 competitor competitor competitor prey prey drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. 8 unspecified role unspecified role bait bait direct interaction direct interaction 3B-2 tnfaip6-cxcl8-4 2 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 3 Supp 1A, Table 1 tnfaip6-cxcl8-5 2 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 4 Supp 1D, Table 1 tnfaip6-cxcl8-6 2 8 unspecified role unspecified role prey prey P10145:p.Arg95Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A Wed May 02 19:48:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg95ala' Tue Jun 30 18:30:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg95Ala' 6 unspecified role unspecified role bait bait drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 3 Supp 1G, Table 1 tnfaip6-cxcl8-7 2 8 unspecified role unspecified role prey prey P10145:p.[Lys91Ala;Lys94Ala;Arg95Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 20:07:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys91ala,lys94ala,arg95ala' Tue Jun 30 19:08:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys91Ala;Lys94Ala;Arg95Ala]' 6 unspecified role unspecified role bait bait drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 3 Supp 1H, Table 1 Table 1 states that the Kd for the CXCL8_T mutant is 2,109;however the image for Supp Fig 1H states it is 2190. tnfaip6-cxcl8-8 2 6 unspecified role unspecified role bait bait link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey direct interaction direct interaction 3 Supp fig 1B, Table 1 cxcl8-tnfaip6-3 2 6 unspecified role unspecified role bait bait link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey direct interaction direct interaction 4 Supp fig 1E, Table 1 cxcl8-tnfaip6-4 2 6 unspecified role unspecified role bait bait link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey P10145:p.Arg95Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A Wed May 02 15:05:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg95ala' Tue Jun 30 20:04:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg95Ala' direct interaction direct interaction 3 Supp fig 1I, Table 1 cxcl8-tnfaip6-5 2 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 3 Supp 1C, Table 1 cxcl8-tnfaip6-6 2 8 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 4 Supp 1F, Table 1 tnfaip6-cxcl8-9 2 9 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait drome-schneider_2 Drosophila melanogaster late embryo cells Fruit fly schneider_2 Established from the late embryo of Drosophila melanogaster Mixture of diploid and tetraploid. Originally had some XY cells now predominantly XX. direct interaction direct interaction 3 Table 1 (row 11) tnfaip6-cxcl8-10 2 6 unspecified role unspecified role bait bait link_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey direct interaction direct interaction 4 Table 1 (row 10)