MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Insulin solubility transitions by pH-dependent interactions with proinsulin C-peptide. Insulin solubility transitions by pH-dependent interactions with proinsulin C-peptide. FEBS J. (1742-464X) 2012 Landreh M., Alvelius G., Willander H., Stukenborg JB., Söder O., Johansson J., Jörnvall H. in vitro In vitro dls dynamic light scattering dls predetermined predetermined participant predetermined Landreh M., Alvelius G., Willander H., Stukenborg JB., Söder O., Johansson J., Jörnvall H. FEBS J. (1742-464X) 2012 Insulin solubility transitions by pH-dependent interactions with proinsulin C-peptide. Insulin solubility transitions by pH-dependent interactions with proinsulin C-peptide. FEBS J. (1742-464X) 2012 Landreh M., Alvelius G., Willander H., Stukenborg JB., Söder O., Johansson J., Jörnvall H. in vitro In vitro fluorescence fluorescence technology fluorescence ms participant Identification by mass spectrometry ms participant Landreh M., Alvelius G., Willander H., Stukenborg JB., Söder O., Johansson J., Jörnvall H. FEBS J. (1742-464X) 2012 Insulin solubility transitions by pH-dependent interactions with proinsulin C-peptide. Insulin solubility transitions by pH-dependent interactions with proinsulin C-peptide. FEBS J. (1742-464X) 2012 Landreh M., Alvelius G., Willander H., Stukenborg JB., Söder O., Johansson J., Jörnvall H. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Landreh M., Alvelius G., Willander H., Stukenborg JB., Söder O., Johansson J., Jörnvall H. FEBS J. (1742-464X) 2012 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 ins-ins-4 2 Insulin 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified C-peptide 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified P01308:p.Glu67Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain E A Tue Jul 12 10:23:41 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 18:39:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu67ala' Tue Jun 30 16:17:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu67Ala' c-peptide C-peptide polyprotein frag polyprotein fragment polyprotein frag certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction homomint mint To determine the pH range for insulin-C-peptide interactions, we used ESI-MS to monitor insulin oligomers between pH 4 to 8 in the presence or absence of equimolar amounts of Cpeptide. In line with previous reports, we observed the highest amount of insulin oligomers at pH 5 [14, 15]. At pH 6, overall poor insulin signals could be detected because of reduced insulin solubility at this pH, and only low amounts of oligomers were observed at higher pH values. The addition of C-peptide removed all insulin oligomers detectable with ESI-MS at pH 5, but had only minor effects at other pH values (Figure 1 and Supplementary Figure 1). We then detected that mixing insulin and C-peptide at pH 5 caused the solution to become visibly turbid. Measurement of the absorbance at 500 nm confirmed that this effect occurred only at pH 5 (Figure 1) and was not influenced by the use of HCl or ammonium acetate (Supplementary Figure 2). The turbidity measurements also indicated that insulin alone is poorly soluble at pH 6, which is compatible with the ESI-MS data. These observations suggest that the removal of insulin oligomers from ESI-MS spectra might be a result of C-peptide-induced insulin precipitation rather than only oligomer dissociation. f1 sf1 sf2 f4 f5 sf5 ins-ins-5 2 Insulin 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified C-peptide 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified c-peptide C-peptide polyprotein frag polyprotein fragment polyprotein frag certain certain sequence position certain certain certain sequence position certain P01308:p.Glu83Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain E A Tue Jul 12 10:23:33 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 16:07:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu83ala' Tue Jun 30 16:21:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu83Ala' direct interaction direct interaction homomint mint To determine the pH range for insulin-C-peptide interactions, we used ESI-MS to monitor insulin oligomers between pH 4 to 8 in the presence or absence of equimolar amounts of Cpeptide. In line with previous reports, we observed the highest amount of insulin oligomers at pH 5 [14, 15]. At pH 6, overall poor insulin signals could be detected because of reduced insulin solubility at this pH, and only low amounts of oligomers were observed at higher pH values. The addition of C-peptide removed all insulin oligomers detectable with ESI-MS at pH 5, but had only minor effects at other pH values (Figure 1 and Supplementary Figure 1). We then detected that mixing insulin and C-peptide at pH 5 caused the solution to become visibly turbid. Measurement of the absorbance at 500 nm confirmed that this effect occurred only at pH 5 (Figure 1) and was not influenced by the use of HCl or ammonium acetate (Supplementary Figure 2). The turbidity measurements also indicated that insulin alone is poorly soluble at pH 6, which is compatible with the ESI-MS data. These observations suggest that the removal of insulin oligomers from ESI-MS spectra might be a result of C-peptide-induced insulin precipitation rather than only oligomer dissociation. f1 sf1 sf2 f4 f5 sf5 ins-ins-6 2 C-peptide 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified P01308:p.Glu83Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain E A Tue Jul 12 10:21:57 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 15:30:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu83ala' Tue Jun 30 17:49:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu83Ala' P01308:p.Glu59Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain E A Tue Jul 12 10:57:52 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:13:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu59ala' Tue Jun 30 16:59:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu59Ala' c-peptide C-peptide polyprotein frag polyprotein fragment polyprotein frag certain certain sequence position certain certain certain sequence position certain P01308:p.Glu67Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain E A Tue Jul 12 10:58:03 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 18:49:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu67ala' Tue Jun 30 19:59:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu67Ala' Insulin 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction homomint mint To determine the pH range for insulin-C-peptide interactions, we used ESI-MS to monitor insulin oligomers between pH 4 to 8 in the presence or absence of equimolar amounts of Cpeptide. In line with previous reports, we observed the highest amount of insulin oligomers at pH 5 [14, 15]. At pH 6, overall poor insulin signals could be detected because of reduced insulin solubility at this pH, and only low amounts of oligomers were observed at higher pH values. The addition of C-peptide removed all insulin oligomers detectable with ESI-MS at pH 5, but had only minor effects at other pH values (Figure 1 and Supplementary Figure 1). We then detected that mixing insulin and C-peptide at pH 5 caused the solution to become visibly turbid. Measurement of the absorbance at 500 nm confirmed that this effect occurred only at pH 5 (Figure 1) and was not influenced by the use of HCl or ammonium acetate (Supplementary Figure 2). The turbidity measurements also indicated that insulin alone is poorly soluble at pH 6, which is compatible with the ESI-MS data. These observations suggest that the removal of insulin oligomers from ESI-MS spectra might be a result of C-peptide-induced insulin precipitation rather than only oligomer dissociation. f1 sf1 sf2 f4 f5 sf5 ins-ins-7 2 Insulin 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified Insulin 4 ms participant Identification by mass spectrometry ms participant experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction homomint mint In line with previous reports, we observed the highest amount of insulin oligomers at pH 5 [14, 15]. At pH 6, overall poor insulin signals could be detected because of reduced insulin solubility at this pH, and only low amounts of oligomers were observed at higher pH values. The addition of C-peptide removed all insulin oligomers detectable with ESI-MS at pH 5, but had only minor effects at other pH values (Figure 1 and Supplementary Figure 1). f1 sf1 sf2 f4 f5 sf5 ins-ins-8 1 C-peptide 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified c-peptide C-peptide polyprotein frag polyprotein fragment polyprotein frag certain certain sequence position certain certain certain sequence position certain Insulin 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f2a homomint mint We therefore conducted DLS analyses at pH 5 and found the presence of oligomers in C-peptide and in insulin solutions, which is in agreement with previous MS studies [13, 26]. In solutions containing an equimolar amount of both peptides, these oligomers could not be observed with DLS (like before not with ESI-MS), but instead large assemblies with a broad size distribution and a hydrodynamic radius exceeding 630 nm (Figure 2). ins-ins-9 1 Insulin 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified Insulin 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f2a homomint mint We therefore conducted DLS analyses at pH 5 and found the presence of oligomers in C-peptide and in insulin solutions, which is in agreement with previous MS studies [13, 26]. In solutions containing an equimolar amount of both peptides, these oligomers could not be observed with DLS (like before not with ESI-MS), but instead large assemblies with a broad size distribution and a hydrodynamic radius exceeding 630 nm (Figure 2). ins-ins-10 3 Insulin 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified C-peptide 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified c-peptide C-peptide polyprotein frag polyprotein fragment polyprotein frag certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2b homomint mint ins-ins-11 3 Insulin 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified Insulin 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f2b homomint mint