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/ Examination of Glycosaminoglycan Binding Sites on the XCL1 Dimer. Examination of Glycosaminoglycan Binding Sites on the XCL1 Dimer. Biochemistry 2016 imex curation Fox JC., Tyler RC., Peterson FC., Dyer DP., Zhang F., Linhardt RJ., Handel TM., Volkman BF. Dark Space = 33? (possibly should not have annotated this publication as no protein- protein interactions other than XCL1 dimerisation - some mutations reduce this. Too late by time I realised) bvolkman@mcw.edu 04-Aug-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps tag fluorescence tag visualisation by fluorescence tag fluorescence Fox JC., Tyler RC., Peterson FC., Dyer DP., Zhang F., Linhardt RJ., Handel TM., Volkman BF. Dark Space = 33? (possibly should not have annotated this publication as no protein- protein interactions other than XCL1 dimerisation - some mutations reduce this. Too late by time I realised) bvolkman@mcw.edu Accepted 2016-JUL-25 AT 11:14 BST AT 11:14 BST by ORCHARD Biochemistry 2016 imex curation I dont know what we would normally use for heparin afiinity columns as I have not come across many/any of these. The pull down definition would appear to include this use-case- "a molecule of interest (bait) is bound to a column, often via an affinity tag expressed as a protein fusion (GST, HIS tag and others) or chemically linked to the bait molecule ." I have altered to "Affinity chrom" anyway.; Monomer of which molecule? Only the XCL1 DIMER binds to Heparin - I left the stoichiometry blank as I could not work out what to enter here. Maybe Heparin should be "2" as well as XCL1, or maybe "1" ? I have entered the CC3 mutant separately as a new interaction, as this is a monomer with 1:1 stoichiometry.-is this what you meant? Examination of Glycosaminoglycan Binding Sites on the XCL1 Dimer. Examination of Glycosaminoglycan Binding Sites on the XCL1 Dimer. Biochemistry 2016 imex curation Fox JC., Tyler RC., Peterson FC., Dyer DP., Zhang F., Linhardt RJ., Handel TM., Volkman BF. Dark Space = 33? (possibly should not have annotated this publication as no protein- protein interactions other than XCL1 dimerisation - some mutations reduce this. Too late by time I realised) bvolkman@mcw.edu 04-Aug-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom predetermined predetermined participant predetermined Fox JC., Tyler RC., Peterson FC., Dyer DP., Zhang F., Linhardt RJ., Handel TM., Volkman BF. Dark Space = 33? (possibly should not have annotated this publication as no protein- protein interactions other than XCL1 dimerisation - some mutations reduce this. Too late by time I realised) Alanine point mutations of several basic residues were made within the W55D background. All variants except CC0, CC3, and WT were made in the W55D background. Several XCL1 variants were eluted from a heparin Sepharose column with increasing NaCl concentrations. To assess the heparin binding affinity for each XCL1 protein, the nonspecific electrostatic attractions measured by cation exchange were subtracted from the values measured by heparin Sepharose chromatography bvolkman@mcw.edu Accepted 2016-JUL-25 AT 11:14 BST AT 11:14 BST by ORCHARD Biochemistry 2016 imex curation I dont know what we would normally use for heparin afiinity columns as I have not come across many/any of these. The pull down definition would appear to include this use-case- "a molecule of interest (bait) is bound to a column, often via an affinity tag expressed as a protein fusion (GST, HIS tag and others) or chemically linked to the bait molecule ." I have altered to "Affinity chrom" anyway.; Monomer of which molecule? Only the XCL1 DIMER binds to Heparin - I left the stoichiometry blank as I could not work out what to enter here. Maybe Heparin should be "2" as well as XCL1, or maybe "1" ? I have entered the CC3 mutant separately as a new interaction, as this is a monomer with 1:1 stoichiometry.-is this what you meant? Examination of Glycosaminoglycan Binding Sites on the XCL1 Dimer. Examination of Glycosaminoglycan Binding Sites on the XCL1 Dimer. Biochemistry 2016 imex curation Fox JC., Tyler RC., Peterson FC., Dyer DP., Zhang F., Linhardt RJ., Handel TM., Volkman BF. Dark Space = 33? (possibly should not have annotated this publication as no protein- protein interactions other than XCL1 dimerisation - some mutations reduce this. Too late by time I realised) bvolkman@mcw.edu 04-Aug-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 Fox JC., Tyler RC., Peterson FC., Dyer DP., Zhang F., Linhardt RJ., Handel TM., Volkman BF. The Kd values obtained were calculated through both kinetic and steady state analysis. Values obtained by each method for each mutant vary considerably. Figures 5A/B appear to show increased binding of the R9A and K25A mutants, but Kd values obtained in Table 2 do not unambiguously support this fact. I have just entered the R9A and K25A mutants as "mutants" as I dont know what category to enter them as - increasing/decreasing/no effect...(maybe I should leave them out altogether?-delete) Dark Space = 33? (possibly should not have annotated this publication as no protein- protein interactions other than XCL1 dimerisation - some mutations reduce this. Too late by time I realised) Alanine point mutations of several basic residues were made within the W55D background. All variants except CC0, CC3, and WT were made in the W55D background. Several XCL1 variants were eluted from a heparin Sepharose with increasing NaCl concentrations. To assess the heparin binding affinity for each XCL1 protein, the nonspecific electrostatic attractions measured by cation exchange were subtracted from the values measured by heparin Sepharose chromatography bvolkman@mcw.edu Accepted 2016-JUL-25 AT 11:14 BST AT 11:14 BST by ORCHARD Biochemistry 2016 imex curation I dont know what we would normally use for heparin afiinity columns as I have not come across many/any of these. The pull down definition would appear to include this use-case- "a molecule of interest (bait) is bound to a column, often via an affinity tag expressed as a protein fusion (GST, HIS tag and others) or chemically linked to the bait molecule ." I have altered to "Affinity chrom" anyway.; Monomer of which molecule? Only the XCL1 DIMER binds to Heparin - I left the stoichiometry blank as I could not work out what to enter here. Maybe Heparin should be "2" as well as XCL1, or maybe "1" ? I have entered the CC3 mutant separately as a new interaction, as this is a monomer with 1:1 stoichiometry.-is this what you meant? hs-10 HS-10 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) heparin-dp4 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) dp8 dp8 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) heparin Heparin small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) p47992-pro_0000005248 Lymphotactin XCL1 LTN SCYC1 ATAC C motif chemokine 1 Cytokine SCM-1 Lymphotaxin SCM-1-alpha Small-inducible cytokine C1 XC chemokine ligand 1 protein protein human Homo sapiens Human VGSEVSDKRTCVSLTTQRLPVSRIKTYTITEGSLRAVIFITKRGLKVCADPQATWVRDVVRSMDRKSNTRNNMIQTKPTGTQQSTNTAVTLTG 22 114 995966DB3438A6A7 hs-15 HS-15 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) hs-14 HS-14 small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) heparin-xcl1-1 2 8 unspecified role unspecified role prey prey P47992-PRO_0000005248:p.Trp55Asp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain W D Wed May 02 16:46:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 17:17:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P47992-PRO_0000005248:p.[Trp55Asp;Arg70Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:05:45 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg70ala,typ55asp' Wed May 02 17:38:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp,arg70ala' Tue Jun 30 17:04:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp55Asp;Arg70Ala]' P47992-PRO_0000005248:p.[Arg9Ala;Trp55Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:00:56 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg9ala,typ55asp' Wed May 02 15:38:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg9ala,trp55asp' Tue Jun 30 17:58:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg9Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Arg18Ala;Trp55Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 10:53:28 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg18ala,typ55asp' Wed May 02 18:31:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg18ala,trp55asp' Tue Jun 30 18:43:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg18Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Arg23Ala;Trp55Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:15:47 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,typ55asp' Wed May 02 15:29:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,trp55asp' Tue Jun 30 18:19:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg23Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Arg35Ala;Trp55Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:12:34 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg35ala,typ55asp' Wed May 02 14:27:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg35ala,trp55asp' Tue Jun 30 19:04:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg35Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Lys42Ala;Trp55Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:05:20 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys42ala,typ55asp' Wed May 02 19:08:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys42ala,trp55asp' Tue Jun 30 18:18:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys42Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Lys25Ala;Trp55Asp] 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 W D Fri Jul 22 10:58:26 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys25ala,typ55asp' Wed May 02 16:05:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys25ala,trp55asp' Tue Jun 30 16:18:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys25Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Arg43Ala;Trp55Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:04:37 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg43ala,typ55asp' Wed May 02 20:00:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg43ala,trp55asp' Tue Jun 30 18:09:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg43Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Lys8Ala;Trp55Asp] 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 W D Tue Jan 24 13:59:54 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'lys8ala,typ55asp' Wed May 02 19:18:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys8ala,trp55asp' Tue Jun 30 17:53:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys8Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Val21Cys;Val59Cys] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V C certain certain sequence position certain certain certain sequence position certain V C Wed May 02 15:48:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val21cys,val59cys' Tue Jun 30 16:20:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val21Cys;Val59Cys]' P47992-PRO_0000005248:p.[Arg23Ala;Arg43Ala;Trp55Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:01:03 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,arg43ala,typ55asp' Wed May 02 16:59:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,arg43ala,trp55asp' Tue Jun 30 19:36:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg23Ala;Arg43Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Lys46Ala;Trp55Asp] 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 W D Thu Jan 26 11:02:49 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'lys46ala,typ55asp' Wed May 02 17:30:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys46ala,trp55asp' Tue Jun 30 16:14:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys46Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Trp55Asp;Arg61Ala] 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 R A certain certain sequence position certain certain certain sequence position certain W D Tue Jan 24 13:46:04 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'arg61ala,typ55asp' Wed May 02 16:04:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp,arg61ala' Tue Jun 30 17:13:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp55Asp;Arg61Ala]' P47992-PRO_0000005248:p.[Cys11Ala;Cys48Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C A certain certain sequence position certain certain certain sequence position certain C A Wed May 02 18:42:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys11ala,cys48ala' Tue Jun 30 17:22:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys11Ala;Cys48Ala]' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait pig Sus scrofa (Pig) Pig direct interaction direct interaction 3, 4A, Table 1 An unfolded/unstructured form of XCL1 showed substantially reduced binding; the CCO mutant has a double mutation of C11 and C48 to alanine which removes an intermolecular disulfide bond. A monomeric form of XCL1 also showed little binding;CC3 is an XCL1 double mutant (V21C, V59C) containing a second disulfide bond which restricts the protein to a monomeric form. heparin-xcl1-2 3 8 unspecified role unspecified role prey prey P47992-PRO_0000005248:p.Trp55Asp 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 W D Wed May 02 17:57:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 16:31:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' P47992-PRO_0000005248:p.[Arg9Ala;Trp55Asp] mutation mutation certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 10:51:13 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg9ala,typ55asp' Wed May 02 16:13:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg9ala,trp55asp' Tue Jun 30 17:52:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg9Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Lys25Ala;Trp55Asp] mutation mutation certain certain sequence position certain certain certain sequence position certain K A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 10:50:10 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys25ala,typ55asp' Wed May 02 18:52:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys25ala,trp55asp' Tue Jun 30 15:43:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys25Ala;Trp55Asp]' P47992-PRO_0000005248:p.[Val21Cys;Val59Cys] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain V C certain certain sequence position certain certain certain sequence position certain V C Wed May 02 17:57:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val21cys,val59cys' Tue Jun 30 18:02:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val21Cys;Val59Cys]' P47992-PRO_0000005248:p.[Arg23Ala;Arg43Ala;Trp55Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 11:01:51 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,arg43ala,typ55asp' Wed May 02 16:22:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,arg43ala,trp55asp' Tue Jun 30 19:51:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg23Ala;Arg43Ala;Trp55Asp]' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait pig Sus scrofa (Pig) Pig direct interaction direct interaction Figure 5, Table 2 A monomeric form of XCL1 (CC3) also showed weaker binding ; CC3 is an XCL1 double mutant (V21C, V59C) containing a second disulfide bond which restricts the protein to a monomeric form. The Kd values obtained were calculated through both kinetic and steady state analysis. Values obtained by each method for each mutant vary considerably. heparin-xcl1-3 1 8 unspecified role unspecified role neutral component neutral component P47992-PRO_0000005248:p.Trp55Asp mutation mutation certain certain sequence position certain certain certain sequence position certain W D Wed May 02 17:03:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 19:41:04 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' P47992-PRO_0000005248:p.[Arg23Ala;Arg43Ala;Trp55Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain W D Fri Jul 22 10:51:05 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,arg43ala,typ55asp' Wed May 02 17:44:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg23ala,arg43ala,trp55asp' Tue Jun 30 19:58:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg23Ala;Arg43Ala;Trp55Asp]' 5 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Figure 6B,7 sodium chloride heparin_dp4-xcl1-1 1 5 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component P47992-PRO_0000005248:p.Trp55Asp mutation mutation certain certain sequence position certain certain certain sequence position certain W D Wed May 02 17:01:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 15:59:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' 5 competitor competitor competitor neutral component neutral component direct interaction direct interaction Figure 6C heparin_dp4-heparin_dp4-1 1 5 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component P47992-PRO_0000005248:p.Trp55Asp mutation mutation certain certain sequence position certain certain certain sequence position certain W D Wed May 02 14:49:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 17:37:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' 6 competitor competitor competitor neutral component neutral component direct interaction direct interaction Figure 6C-2 heparin_dp4-heparin_dp4-2 1 5 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component P47992-PRO_0000005248:p.Trp55Asp mutation mutation certain certain sequence position certain certain certain sequence position certain W D Wed May 02 15:11:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 17:51:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' 4 competitor competitor competitor neutral component neutral component direct interaction direct interaction Figure 7 An unsulfated heparan molecule and a less sulfated molecule were unable to compete for the binding sites occupied by the dp4 probe. heparin_dp4-hs_10-1 1 5 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component P47992-PRO_0000005248:p.Trp55Asp mutation mutation certain certain sequence position certain certain certain sequence position certain W D Wed May 02 19:35:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 16:16:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' 10 competitor competitor competitor neutral component neutral component direct interaction direct interaction Figure 7-2 An unsulfated heparan molecule and a less sulfated molecule were unable to compete for the binding sites occupied by the dp4 probe. heparin_dp4-hs_14-1 1 5 unspecified role unspecified role neutral component neutral component region fluorescein fluorescein label fluorescein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component P47992-PRO_0000005248:p.Trp55Asp mutation mutation certain certain sequence position certain certain certain sequence position certain W D Wed May 02 18:27:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp55asp' Tue Jun 30 18:24:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp55Asp' 9 competitor competitor competitor neutral component neutral component direct interaction direct interaction Figure 7-3 An unsulfated heparan molecule and a less sulfated molecule were unable to compete for the binding sites occupied by the dp4 probe. heparin-xcl1-4 3 8 unspecified role unspecified role prey prey P47992-PRO_0000005248:p.[Val21Cys;Val59Cys] mutation mutation certain certain sequence position certain certain certain sequence position certain V C certain certain sequence position certain certain certain sequence position certain V C Wed May 02 14:41:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val21cys,val59cys' Tue Jun 30 19:49:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val21Cys;Val59Cys]' ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait pig Sus scrofa (Pig) Pig direct interaction direct interaction Figure 5i, Table 2 A monomeric form of XCL1 (CC3) also showed weaker binding ; CC3 is an XCL1 double mutant (V21C, V59C) containing a second disulfide bond which restricts the protein to a monomeric form. The Kd values obtained were calculated through both kinetic and steady state analysis. Values obtained by each method for each mutant vary considerably.