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/ Dual functionality of beta-tryptase protomers as both proteases and cofactors in the active tetramer. Dual functionality of beta-tryptase protomers as both proteases and cofactors in the active tetramer. The Journal of biological chemistry 2018 imex curation Maun HR., Liu PS., Franke Y., Eigenbrot C., Forrest WF., Schwartz LB., Lazarus RA. Beta-Tryptase enzymatic assays using the synthetic substrate S-2288 (Fig.1B-C, Fig.4C, Table2, Fig.7, Fig.9-11) were not annotated since they are not biologically relevant interactions. Interactions between Beta-Triptase and the B12 anti-tryptase monoclonal antibody (Fig. 6) were not annotate. lazarus.bob@gene.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Maun HR., Liu PS., Franke Y., Eigenbrot C., Forrest WF., Schwartz LB., Lazarus RA. Beta-Tryptase enzymatic assays using the synthetic substrate S-2288 (Fig.1B-C, Fig.4C, Table2, Fig.7, Fig.9-11) were not annotated since they are not biologically relevant interactions. Interactions between Beta-Triptase and the B12 anti-tryptase monoclonal antibody (Fig. 6) were not annotate. lazarus.bob@gene.com Accepted 2018-OCT-09 AT 13:54 BST AT 13:54 BST by LUANA The Journal of biological chemistry 2018 imex curation Dual functionality of beta-tryptase protomers as both proteases and cofactors in the active tetramer. Dual functionality of beta-tryptase protomers as both proteases and cofactors in the active tetramer. The Journal of biological chemistry 2018 imex curation Maun HR., Liu PS., Franke Y., Eigenbrot C., Forrest WF., Schwartz LB., Lazarus RA. Beta-Tryptase enzymatic assays using the synthetic substrate S-2288 (Fig.1B-C, Fig.4C, Table2, Fig.7, Fig.9-11) were not annotated since they are not biologically relevant interactions. Interactions between Beta-Triptase and the B12 anti-tryptase monoclonal antibody (Fig. 6) were not annotate. lazarus.bob@gene.com 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro comigration in sds comigration in sds page comigration in sds weight by comassie molecular weight estimation by coomasie staining weight by comassie Maun HR., Liu PS., Franke Y., Eigenbrot C., Forrest WF., Schwartz LB., Lazarus RA. SDS page under nonreducing (NR) conditions. Beta-Tryptase enzymatic assays using the synthetic substrate S-2288 (Fig.1B-C, Fig.4C, Table2, Fig.7, Fig.9-11) were not annotated since they are not biologically relevant interactions. Interactions between Beta-Triptase and the B12 anti-tryptase monoclonal antibody (Fig. 6) were not annotate. lazarus.bob@gene.com Accepted 2018-OCT-09 AT 13:54 BST AT 13:54 BST by LUANA The Journal of biological chemistry 2018 imex curation Dual functionality of beta-tryptase protomers as both proteases and cofactors in the active tetramer. Dual functionality of beta-tryptase protomers as both proteases and cofactors in the active tetramer. The Journal of biological chemistry 2018 imex curation Maun HR., Liu PS., Franke Y., Eigenbrot C., Forrest WF., Schwartz LB., Lazarus RA. Beta-Tryptase enzymatic assays using the synthetic substrate S-2288 (Fig.1B-C, Fig.4C, Table2, Fig.7, Fig.9-11) were not annotated since they are not biologically relevant interactions. Interactions between Beta-Triptase and the B12 anti-tryptase monoclonal antibody (Fig. 6) were not annotate. lazarus.bob@gene.com 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 predetermined predetermined participant predetermined Maun HR., Liu PS., Franke Y., Eigenbrot C., Forrest WF., Schwartz LB., Lazarus RA. in this case I've used the full gene sequence (otherwise there are mutations that cannot be mapped). To map the mutations I've used Supplementary Figure2. Beta-Tryptase enzymatic assays using the synthetic substrate S-2288 (Fig.1B-C, Fig.4C, Table2, Fig.7, Fig.9-11) were not annotated since they are not biologically relevant interactions. Interactions between Beta-Triptase and the B12 anti-tryptase monoclonal antibody (Fig. 6) were not annotate. lazarus.bob@gene.com Accepted 2018-OCT-09 AT 13:54 BST AT 13:54 BST by LUANA The Journal of biological chemistry 2018 imex curation tryb1_human Tryptase alpha/beta-1 TPSAB1 TPSB1 Tryptase I TPS1 TPS2 Tryptase alpha-1 protein protein human Homo sapiens Human MLNLLLLALPVLASRAYAAPAPGQALQRVGIVGGQEAPRSKWPWQVSLRVHGPYWMHFCGGSLIHPQWVLTAAHCVGPDVKDLAALRVQLREQHLYYQDQLLPVSRIIVHPQFYTAQIGADIALLELEEPVNVSSHVHTVTLPPASETFPPGMPCWVTGWGDVDNDERLPPPFPLKQVKVPIMENHICDAKYHLGAYTGDDVRIVRDDMLCAGNTRRDSCQGDSGGPLVCKVNGTWLQAGVVSWGEGCAQPNRPGIYTRVTYYLDWIHHYVPKKP ADC48FDC51F37112 q15661-pro_0000027480 Tryptase alpha/beta-1 TPSAB1 TPS1 TPS2 TPSB1 Tryptase I Tryptase alpha-1 protein protein human Homo sapiens Human IVGGQEAPRSKWPWQVSLRVHGPYWMHFCGGSLIHPQWVLTAAHCVGPDVKDLAALRVQLREQHLYYQDQLLPVSRIIVHPQFYTAQIGADIALLELEEPVNVSSHVHTVTLPPASETFPPGMPCWVTGWGDVDNDERLPPPFPLKQVKVPIMENHICDAKYHLGAYTGDDVRIVRDDMLCAGNTRRDSCQGDSGGPLVCKVNGTWLQAGVVSWGEGCAQPNRPGIYTRVTYYLDWIHHYVPKKP 31 275 C52ADDBCC83FE6B7 beta-tryptase-1 1 5 unspecified role unspecified role neutral component neutral component purified purified Q15661-PRO_0000027480:p.Ile88Cys I99C mutation mutation certain certain sequence position certain certain certain sequence position certain I C Tue Jun 30 17:46:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile88Cys' spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction Table 3, Fig.5 2.72A Rworking 0.193 Rfree 0.227 in this case I've used the post-processed chain. To map the mutation I've used Supplementary Figure2, In this case I99C corresponds to I118C in the full gene sequence, in turn it corresponds to I88C in the post-processed chain. tpsab1-2 3 4 unspecified role unspecified role neutral component neutral component purified purified Q15661:p.Tyr97Cys Y75C mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain Y C Tue Jun 30 19:39:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr97Cys' Q15661:p.Ile118Cys I99C mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain I C Tue Jun 30 17:44:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile118Cys' binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain Q15661:p.Ile31Gly I16G mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain I G Tue Jun 30 18:47:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile31Gly' Q15661:p.[Tyr54Ala;Tyr97Cys;Ile118Cys] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y C certain certain sequence position certain certain certain sequence position certain I C certain certain sequence position certain certain certain sequence position certain Y A Wed Sep 26 11:56:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr97Cys;Ile118Cys;Tyr54Ala]' Tue Jun 30 16:47:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr54Ala;Tyr97Cys;Ile118Cys]' trini Trichoplusia ni (Cabbage looper) Cabbage looper direct interaction direct interaction Fig.2, Fig.4A, Fig.8 As shown in Fig. 2, WT Beta-tryptase migrates as a tetramer. However, the I16G N-terminal insertion mutant does not, instead migrating as a monomer with a comparable retention time to the Beta-tryptase zymogen [...] Serendipitously, we found monomeric fractions that were cysteinylated at I99C (I99C*), which prevents dimerization with another -tryptase monomer via the large interface, whereas the mutations Y75A and Y37bA abrogate interactions via the small interface, thus preventing tetramer formation. tpsab1-1 3 4 unspecified role unspecified role neutral component neutral component purified purified Q15661:p.Tyr97Cys Y75C mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain Y C Tue Jun 30 18:43:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr97Cys' Q15661:p.Ile118Cys I99C mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain I C Tue Jun 30 15:35:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile118Cys' binding region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain cysteinylated residue uncategorized protein modification uncategorized protein modification certain certain sequence position certain certain certain sequence position certain disrupting-ptm ptm disrupting an interaction disrupting-ptm Q15661:p.[Tyr54Ala;Tyr97Cys;Ile118Cys] I99C/Y75A/Y37bA mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain Y C certain certain sequence position certain certain certain sequence position certain I C certain certain sequence position certain certain certain sequence position certain Y A Wed Sep 26 11:21:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr97Cys;Ile118Cys;Tyr54Ala]' Tue Jun 30 19:04:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr54Ala;Tyr97Cys;Ile118Cys]' trini Trichoplusia ni (Cabbage looper) Cabbage looper direct interaction direct interaction Fig.4A, Fig.8 we engineered a dimer at the large interface (I99C/Y75A/Y37bA) unable to form a tetramer based on SEC (Fig. 8)[...]Serendipitously, we found monomeric fractions that were cysteinylated at I99C (I99C*), which prevents dimerization with another -tryptase monomer via the large interface, whereas the mutations Y75A and Y37bA abrogate interactions via the small interface, thus preventing tetramer formation. heparin stabilises the dimer tpsab1-3 2 4 unspecified role unspecified role neutral component neutral component purified purified Q15661:p.Tyr97Cys Y75C mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain Y C Tue Jun 30 20:14:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr97Cys' Q15661:p.Ile118Cys I99C mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain I C Tue Jun 30 16:22:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile118Cys' trini Trichoplusia ni (Cabbage looper) Cabbage looper disulfide bond disulfide bond disulfide bridge SS-bond Fig.4B Analysis of tetramers by SDS-PAGE under nonreducing conditions showed WT migrating as a monomer, whereas the Y75C and I99C mutants migrated as dimers (Fig. 4B). These dimers collapsed to the monomers when analyzed by SDS-PAGE under reducing conditions, consistent with an intermolecular disulfide bond between the two monomers