UniProt UniProt http://www.uniprot.org Identification of Human Islet Amyloid Polypeptide as a BACE2 Substrate. Identification of Human Islet Amyloid Polypeptide as a BACE2 Substrate. PloS one 2015 imex curation Rulifson IC., Cao P., Miao L., Kopecky D., Huang L., White RD., Samayoa K., Gardner J., Wu X., Chen K., Tsuruda T., Homann O., Baribault H., Yamane H., Carlson T., Wiltzius J., Li Y. ingridr@amgen.com 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protease assay protease assay ms participant Identification by mass spectrometry ms participant Rulifson IC., Cao P., Miao L., Kopecky D., Huang L., White RD., Samayoa K., Gardner J., Wu X., Chen K., Tsuruda T., Homann O., Baribault H., Yamane H., Carlson T., Wiltzius J., Li Y. IAPP amino acids renumbered to full length sequence ingridr@amgen.com Accepted 2019-MAY-23 AT 08:29 BST AT 08:29 BST by LIVIAP PloS one 2015 Only protein-protein interactions imex curation imex curation In general you have added all the peptides as sufficient to bind, however, as far as I could see these are the clevage product... we can discuss it. Sandra - No they aren't - this is the full-length peptide as described in M&M mapped to the protein sequence Please add a data comment explaining why all your mutants are shifted. Sandra - I have added this, but should point out we don't normally do so - apart from anything else, it cannot be automatically updated if the sequence changes. Please add the cleavage sites as resulting PTMs. Sandra - how? We don't have a feature type that works for this (or if we do, I can't find it); peptide sequence given in M&M, this was mapped to full length protein. Identification of Human Islet Amyloid Polypeptide as a BACE2 Substrate. Identification of Human Islet Amyloid Polypeptide as a BACE2 Substrate. PloS one 2015 imex curation Rulifson IC., Cao P., Miao L., Kopecky D., Huang L., White RD., Samayoa K., Gardner J., Wu X., Chen K., Tsuruda T., Homann O., Baribault H., Yamane H., Carlson T., Wiltzius J., Li Y. ingridr@amgen.com 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro aggregation aggregation assay aggregation tag fluorescence tag visualisation by fluorescence tag fluorescence Rulifson IC., Cao P., Miao L., Kopecky D., Huang L., White RD., Samayoa K., Gardner J., Wu X., Chen K., Tsuruda T., Homann O., Baribault H., Yamane H., Carlson T., Wiltzius J., Li Y. Interaction assayed via incorporation of Thioflavin T (ThT), a spectroscopic dye whose fluorometric properties change upon binding to β-sheet rich amyloid fibrils ingridr@amgen.com Accepted 2019-MAY-23 AT 08:29 BST AT 08:29 BST by LIVIAP PloS one 2015 Only protein-protein interactions imex curation imex curation In general you have added all the peptides as sufficient to bind, however, as far as I could see these are the clevage product... we can discuss it. Sandra - No they aren't - this is the full-length peptide as described in M&M mapped to the protein sequence Please add a data comment explaining why all your mutants are shifted. Sandra - I have added this, but should point out we don't normally do so - apart from anything else, it cannot be automatically updated if the sequence changes. Please add the cleavage sites as resulting PTMs. Sandra - how? We don't have a feature type that works for this (or if we do, I can't find it); peptide sequence given in M&M, this was mapped to full length protein. iapp_human Islet amyloid polypeptide IAPP Amylin Diabetes-associated peptide Insulinoma amyloid peptide protein protein human Homo sapiens Human MGILKLQVFLIVLSVALNHLKATPIESHQVEKRKCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTYGKRNAVEVLKREPLNYLPL DIP protein P10997 original sequence version: 138 not sure about the protein origin DIP protein P10997 original sequence version: 92 DIP protein P10997 original sequence version: 88 mint AA8B1F7FD9FCB4BD ins_human Insulin INS protein protein human Homo sapiens Human MALWMRLLPLLALLALWGPDPAAAFVNQHLCGSHLVEALYLVCGERGFFYTPKTRREAEDLQVGQVELGGGPGAGSLQPLALEGSLQKRGIVEQCCTSICSLYQLENYCN DIP protein P01308 original sequence version: 184 DIP protein P01308 original sequence version: 104 DIP protein P01308 original sequence version: 177 Insulin decreases blood glucose concentration. It increases cell permeability to monosaccharides, amino acids and fatty acids. It accelerates glycolysis, the pentose phosphate cycle, and glycogen synthesis in liver mint homomint Defects in INS are the cause of familial hyperproinsulinemia [MIM:176730] C2C3B23B85E520E5 bace1_human Beta-secretase 1 BACE KIAA1149 Beta-site amyloid precursor protein cleaving enzyme 1 Membrane-associated aspartic protease 2 Memapsin-2 Aspartyl protease 2 BACE1 protein protein human Homo sapiens Human MAQALPWLLLWMGAGVLPAHGTQHGIRLPLRSGLGGAPLGLRLPRETDEEPEEPGRRGSFVEMVDNLRGKSGQGYYVEMTVGSPPQTLNILVDTGSSNFAVGAAPHPFLHRYYQRQLSSTYRDLRKGVYVPYTQGKWEGELGTDLVSIPHGPNVTVRANIAAITESDKFFINGSNWEGILGLAYAEIARPDDSLEPFFDSLVKQTHVPNLFSLQLCGAGFPLNQSEVLASVGGSMIIGGIDHSLYTGSLWYTPIRREWYYEVIIVRVEINGQDLKMDCKEYNYDKSIVDSGTTNLRLPKKVFEAAVKSIKAASSTEKFPDGFWLGEQLVCWQAGTTPWNIFPVISLYLMGEVTNQSFRITILPQQYLRPVEDVATSQDDCYKFAISQSSTGTVMGAVIMEGFYVVFDRARKRIGFAVSACHVHDEFRTAAVEGPFVTLDMEDCGYNIPQTDESTLMTIAYVMAAICALFMLPLCLMVCQWRCLRCLRQQHDDFADDISLLK DIP protein P56817 original sequence version: 161 377CE4C824ACEF05 377CE4C824ACEF05 bace2_human Beta-secretase 2 BACE2 AEPLC ALP56 ASP21 Aspartic-like protease 56 kDa Aspartyl protease 1 Beta-site amyloid precursor protein cleaving enzyme 2 Down region aspartic protease Memapsin-1 Membrane-associated aspartic protease 1 Theta-secretase CDA13 UNQ418/PRO852 protein protein human Homo sapiens Human MGALARALLLPLLAQWLLRAAPELAPAPFTLPLRVAAATNRVVAPTPGPGTPAERHADGLALALEPALASPAGAANFLAMVDNLQGDSGRGYYLEMLIGTPPQKLQILVDTGSSNFAVAGTPHSYIDTYFDTERSSTYRSKGFDVTVKYTQGSWTGFVGEDLVTIPKGFNTSFLVNIATIFESENFFLPGIKWNGILGLAYATLAKPSSSLETFFDSLVTQANIPNVFSMQMCGAGLPVAGSGTNGGSLVLGGIEPSLYKGDIWYTPIKEEWYYQIEILKLEIGGQSLNLDCREYNADKAIVDSGTTLLRLPQKVFDAVVEAVARASLIPEFSDGFWTGSQLACWTNSETPWSYFPKISIYLRDENSSRSFRITILPQLYIQPMMGAGLNYECYRFGISPSTNALVIGATVMEGFYVIFDRAQKRVGFAASPCAEIAGAAVSEISGPFSTEDVASNCVPAQSLSEPILWIVSYALMSVCGAILLVLIVLLLLPFRCQRRPRDPEVVNDESSLVRHRWK 2E903150823760D3 iapp_mouse Islet amyloid polypeptide Amylin Diabetes-associated peptide Iapp protein protein mouse Mus musculus Mouse MMCISKLPAVLLILSVALNHLRATPVRSGSNPQMDKRKCNTATCATQRLANFLVRSSNNLGPVLPPTNVGSNTYGKRNAAGDPNRESLDFLLV B135DBBC81475B15 bace2-iapp-1 1 3 enzyme target enzyme target substrate substrate neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P10997:p.[Phe48Lys;Phe56Lys] mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain F K certain certain sequence position certain certain certain sequence position certain F K Fri Feb 01 16:39:35 GMT 2019 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe48Lys;Phe56Lys' Tue Jun 30 16:48:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe48Lys;Phe56Lys]' P10997:p.Phe48Lys mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain F K Tue Jun 30 17:23:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe48Lys' P10997:p.Phe56Lys mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain F K Tue Jun 30 17:38:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe56Lys' chemical synthesis Chemical synthesis (Chemical synthesis) 6 enzyme enzyme neutral component neutral component purified purified region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. protein cleavage protein cleavage Fig. 2B, 3B, 3E, 3H To determine if recombinant human BACE2 is sufficient to proteolytically cleave human IAPP peptide (hIAPP), we used mass spectrometry (MS) to identify hIAPP fragments resulting from BACE2 digestion in vitro. bace1-iapp-1 1 3 enzyme target enzyme target substrate substrate neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P10997:p.[Phe48Lys;Phe56Lys] mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain F K certain certain sequence position certain certain certain sequence position certain F K Fri Feb 01 16:07:42 GMT 2019 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe48Lys;Phe56Lys' Tue Jun 30 19:10:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Phe48Lys;Phe56Lys]' P10997:p.Phe48Lys mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain F K Tue Jun 30 19:04:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe48Lys' P10997:p.Phe56Lys 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 K Tue Jun 30 19:28:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe56Lys' chemical synthesis Chemical synthesis (Chemical synthesis) 5 enzyme enzyme neutral component neutral component purified purified P56817:p.Glu46Ser mutation mutation certain certain sequence position certain certain certain sequence position certain E S Tue Jun 30 18:13:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu46Ser' region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. protein cleavage protein cleavage Fig. 2C, 3C, 3F, 3I bace1-iapp-2 1 7 enzyme target enzyme target substrate substrate neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 5 enzyme enzyme neutral component neutral component purified purified P56817:p.Glu46Ser mutation mutation certain certain sequence position certain certain certain sequence position certain E S Tue Jun 30 19:26:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu46Ser' region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. protein cleavage protein cleavage Fig. 4C bace2-iapp-2 1 6 enzyme enzyme neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 7 enzyme target enzyme target substrate substrate neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) protein cleavage protein cleavage Fig. 4B bace2-iapp-3 1 4 inhibitor inhibitor neutral component neutral component 3 enzyme target enzyme target substrate substrate neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 6 enzyme enzyme neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. protein cleavage protein cleavage Fig. 5 Only mature insulin acted as an inhibitor, not the propeptide iapp-iapp-5 2 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Fig 6. The kinetics of hIAPP fibril formation as a function of increasing recombinant BACE concentration was determined in vitro via incorporation of Thioflavin T (ThT), a spectroscopic dye whose fluorometric properties change upon binding to β-sheet rich amyloid fibrils [45]. iapp-iapp-6 2 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 5 inhibitor inhibitor neutral component neutral component purified purified crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. direct interaction direct interaction Fig 6. The addition of recombinant BACE1 to hIAPP in the ThT assay showed a dose-dependent delay in the rate of hIAPP fibril formation (Fig 6A), bace2-iapp-4 2 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 6 inhibitor inhibitor neutral component neutral component purified purified crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. direct interaction direct interaction Fig 6. When BACE2 was added to hIAPP in the ThT assay, however, a more robust impairment of fibril formation was observed,