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/ Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro cd circular dichroism CD CD cd predetermined predetermined participant predetermined Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: 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 Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight identificat confirmation by molecular weight weight identificat Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. The hSLBP-hSLIP1 complex was co-expressed in bacteria. Full-length hSLBP as well as hSLBP-ND were in a co-expression system in which only hSLIP1 was (His)6-tagged. Full-length hSLBP formed a very stable complex with hSLIP1 and was copurified with hSLIP1 over a nickel affinity column as well as a gel filtration column. The proteins form a stoichiometric complex as visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and were confirmed by Western blotting. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro atomic force microsc atomic force microscopy AFM AFM atomic force microsc predetermined predetermined participant predetermined Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. Analytical unltracentrifugation (AUC) was used on the mix of purified interacting proteins prior to analysis using atomic force microscope (AFM). rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column sequence tag sequence tag identification sequence tag Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column fingerprinting peptide massfingerprinting fingerprinting Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Assembly of the SLIP1-SLBP complex on histone mRNA requires heterodimerization and sequential binding of SLBP followed by SLIP1. Biochemistry 2013 imex curation Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R., Bansal N., Zhang M., Bhaskar A., Itotia P., Lee E., Shlyakhtenko LS., Lam TT., Fritz A., Berezney R., Lyubchenko YL., Stafford WF., Thapar R. rthapar@hwi.buffalo.edu Accepted 2018-DEC-03 AT 16:09 GMT AT 16:09 GMT by PPORRAS Biochemistry 2013 Only protein-protein interactions imex curation imex curation Not sure about expt prep :(; 'Expressed in' field is missing in many instances. Some proteins have been expressed in baculovirus, essentially using an insect system (check appropriate reference). --> Spofr. Reference to figure 1 does not apply. --> EBI-20718186: 1c, 1d. Please correct "fig.s" with "FIgs.", otherwise it looks like you are pointing to a supplementary figure. —> done. Add exp modification comment stating how AUC was used in combination with AFM. —> Can’t see where I need to add anything. Also, phosphorylated proteins need to have this annotated as a feature. EBI-20718186: unphosphorylated EBI-20718261: unphosphorylated EBI-20718267: unphosphorylated EBI-20719652: phosphorylation there already EBI-20719661: phosphorylation there already EBI-20718191: phosphorylation added Also changed the purification method to "purified" as for other expts. ; Looking at the maps again, there are vectors with HA tags at N, C and both termini. Change to "?-?" ?; As before, exp prep is not 'over-expressed', but in this case 'partially purified'. Because of this, I think interaction type needs to be 'physical association' in this case. --> how it this different here? The text states that "We expressed full-length hSLBP as well as hSLBP-ND in a co-expression system wherein only hSLIP1 was (His)6-tagged", so I think slbp his tag feature needs to be removed. --> agree, removed. Please add an exp-modification annotation at experiment level stating how the proteins were co-expressed and partially co-purified through nickel chromatography before being passed through a gel filtration column for interaction detection. --> done. Mutants from figure 7c should be curated here as feature. --> added. ; All comments got copied when I cloned the interaction into this new expt... Expt created on request. cloned from EBI-20718250. ; As before, exp-prep problem. --> changed. I am not sure if in this exp they used the result from exp in figure 4a, in which case the proteins would be partially purified and the interaction type cannot be direct. --> I agree, it looks like they used the same prep as for 4a but they don't spell it out. Do we need to add a comment or caution? NEW: FL-SLBP is phosphorylised, not His-tagged - I made that change now. AUC experiments from figure 4D are missing, the relevant interaction detection method is 'cosedimentation in solution' (in drop down menu, 'solution sedimentati'). --> new expt: -8, interaction EBI-20978866. ; Exp prep as before. --> done. Kds in specific mutants can be captured as 'feature parameters' in each case. --> done. Co-purification exp reported in figure 7c is the same as molecular sieving +wb experiment, so the mutant and the ref to the figure legend should go there. --> done, although 202 was also tested by SPR so mutant stays here, too. Please add Table 2 in fig legend as the source of the Kds. --> done. ; Same exp prep problem as previously. --> exp prep changed and expr-in added for F5b. Exps from figures 5a and 5b should be different: 5a is a maldi-tof, so part det met is 'peptide massfinger printing'. 5b is an electrospray (Qtof), so part det met is 'sequence tag'. --> part det changed and 3 interactions from 5b moved to new expt -9.; Comments were copied from publ level (see expt 8...). Moved the interactions for F5b from expt 6 to this one as part det diff. m4gdb_danre MIF4G domain-containing protein B mif4gdb zgc:110826 protein protein danre Danio rerio (Zebrafish) Brachydanio rerio Zebrafish MENSSKEDYKIQSFDLETQKLLKTALKDPGSVDLEKVSSVIVDQSLKDQVFSREAGRICYTIVQAEAKQTNGSVFRRNLLNRLQQEFKAREETRKRSTQEWVCLVSFICNIFDYLKVNNMPMVALVHPVYDCLFRLAQSDALKNEEEVDCLVLQLHRIGDQLEKMNVQLMDELFNLLRDGFLLQEDLSSMGRLLLLEILEFRAGGWKLSDTAQKYYYSEVTD 26059435E7469867 mi4gd_human MIF4G domain-containing protein SLIP1 SLBP-interacting protein 1 MIF4GD protein protein human Homo sapiens Human MGEPSREEYKIQSFDAETQQLLKTALKDPGAVDLEKVANVIVDHSLQDCVFSKEAGRMCYAIIQAESKQAGQSVFRRGLLNRLQQEYQAREQLRARSLQGWVCYVTFICNIFDYLRVNNMPMMALVNPVYDCLFRLAQPDSLSKEEEVDCLVLQLHRVGEQLEKMNGQRMDELFVLIRDGFLLPTGLSSLAQLLLLEIIEFRAAGWKTTPAAHKYYYSEVSD 07E5B651D638AC9D slbp_human Histone RNA hairpin-binding protein SLBP HBP Histone stem-loop-binding protein protein protein human Homo sapiens Human MACRPRSPPRHQSRCDGDASPPSPARWSLGRKRRADGRRWRPEDAEEAEHRGAERRPESFTTPEGPKPRSRCSDWASAVEEDEMRTRVNKEMARYKRKLLINDFGRERKSSSGSSDSKESMSTVPADFETDESVLMRRQKQINYGKNTIAYDRYIKEVPRHLRQPGIHPKTPNKFKKYSRRSWDQQIKLWKVALHFWDPPAEEGCDLQEIHPVDLESAESSSEPQTSSQDDFDVYSGTPTKVRHMDSQVEDEFDLEACLTEPLRDFSAMS DIP protein Q14493 original sequence version: 119 DIP protein Q14493-1 original sequence version: 139 4E84E502393D1BF7 mif4gd-1 3 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1a (left), 4a mif4gdb-1 3 10 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1a (right) mif4gd-3 3 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1a (left) mif4gdb-2 3 10 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1a (right) mif4gd-4 6 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction Figs 1c, 1d, 6a, 6b slbp-1 6 12 unspecified role unspecified role neutral component neutral component purified purified phosph171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm 12 unspecified role unspecified role neutral component neutral component purified purified phosph171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction Figs. 2g, 2h mif4gd-slbp-1 9 12 unspecified role unspecified role neutral component neutral component purified purified n-terminal domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain radiolabel 15N 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined his tag 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 ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figs 2f, S3b, S3c mif4gd-slbp-2 4 12 unspecified role unspecified role neutral component neutral component partially purified partially purified 111-270 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role neutral component neutral component partially purified partially purified his tag 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 A9UHW6:p.Arg202Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain R A Tue Nov 27 16:34:19 GMT 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'p.arg202ala' Tue Jun 30 19:17:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg202Ala' ecoli Escherichia coli (strain K12) physical association physical association Fig. 4a SLBP cannot be expressed in bacteria on its own. Here, it has been co-expressed in ecoli with MIF4GD. mif4gd-slbp-3 1 12 unspecified role unspecified role neutral component neutral component partially purified partially purified phospho171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role neutral component neutral component partially purified partially purified his tag 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 4b SLBP cannot be expressed in bacteria on its own. Here, it has been co-expressed in ecoli with MIF4GD. mif4gd-slbp-4 6 12 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figs 6a, 6b mif4gd-slbp-5 6 12 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figs 6a, 6b slbp-3 8 12 unspecified role unspecified role neutral component neutral component purified purified 111-270 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5a slbp-4 8 12 unspecified role unspecified role neutral component neutral component purified purified 111-270 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5a slbp-5 8 12 unspecified role unspecified role neutral component neutral component purified purified 111-270 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5a slbp-6 8 12 unspecified role unspecified role neutral component neutral component purified purified 111-270 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5a slbp-7 7 12 unspecified role unspecified role neutral component neutral component purified purified 111-270 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain phospho171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction Fig. 5b slbp-8 7 12 unspecified role unspecified role neutral component neutral component purified purified 111-270 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain phospho171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction Fig. 5b slbp-9 7 12 unspecified role unspecified role neutral component neutral component purified purified 111-270 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain phospho171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction Fig. 5b mif4gd-slbp-6 6 12 unspecified role unspecified role neutral component neutral component purified purified phosph171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figs 6c, 6d mif4gd-slbp-7 6 12 unspecified role unspecified role neutral component neutral component purified purified phosph171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm 11 unspecified role unspecified role neutral component neutral component purified purified his tag 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figs 6c, 6d mif4gd-slbp-8 2 12 unspecified role unspecified role bait bait purified purified phosph171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain 11 unspecified role unspecified role prey prey purified purified A9UHW6:p.Glu7Ala 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 E A Tue Jun 30 16:50:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu7Ala' A9UHW6:p.Lys10Ala 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 Tue Jun 30 20:21:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys10Ala' his tag 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 A9UHW6:p.Arg202Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 16:32:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg202Ala' A9UHW6:p.Arg57Ala 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 Tue Jun 30 15:42:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg57Ala' A9UHW6:p.Tyr60Ala 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 Y A Tue Jun 30 16:50:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr60Ala' A9UHW6:p.Gln64Ala 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 Q A Tue Jun 30 16:45:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln64Ala' A9UHW6:p.Arg76Ala 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 Tue Jun 30 18:19:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg76Ala' A9UHW6:p.Arg157Ala 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 Tue Jun 30 16:31:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg157Ala' A9UHW6:p.His44Ala 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 H A Tue Jun 30 16:33:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His44Ala' A9UHW6:p.Phe75Ala 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 F A Tue Jun 30 20:02:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe75Ala' unspecified mutation 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 This feature was amended as a result of our quality control procedures. The original short label was 'p.Phe87Ala'. A9UHW6:p.Arg82Ala 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 Tue Jun 30 16:29:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg82Ala' unspecified mutation 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 This mutation was emended following our quality control procedures. The original short label was 'p.Arg93Ala'. A9UHW6:p.Glu200Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 18:12:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu200Ala' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 7d, Table 2 Mutant mif4gd trp206ala resulted in an unstable construct and was not tested. mif4gd-slbp-10 5 12 unspecified role unspecified role neutral component neutral component partially purified partially purified phospho171 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role neutral component neutral component partially purified partially purified his tag 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 ecoli Escherichia coli (strain K12) physical association physical association Fig. 4d SLBP cannot be expressed in bacteria on its own. Here, it has been co-expressed in ecoli with MIF4GD.