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 Amino acid sequence determinants in self-assembly of insulin chiral amyloid superstructures: role of C-terminus of B-chain in association of fibrils. Amino acid sequence determinants in self-assembly of insulin chiral amyloid superstructures: role of C-terminus of B-chain in association of fibrils. FEBS Lett. (0014-5793) 2013 Babenko V., Dzwolak W. in vitro In vitro ir spectrometry infrared spectroscopy ir spectrometry predetermined predetermined participant predetermined Babenko V., Dzwolak W. FEBS Lett. (0014-5793) 2013 Amino acid sequence determinants in self-assembly of insulin chiral amyloid superstructures: role of C-terminus of B-chain in association of fibrils. Amino acid sequence determinants in self-assembly of insulin chiral amyloid superstructures: role of C-terminus of B-chain in association of fibrils. FEBS Lett. (0014-5793) 2013 Babenko V., Dzwolak W. in vitro In vitro atomic force microsc atomic force microscopy AFM AFM atomic force microsc predetermined predetermined participant predetermined Babenko V., Dzwolak W. FEBS Lett. (0014-5793) 2013 Amino acid sequence determinants in self-assembly of insulin chiral amyloid superstructures: role of C-terminus of B-chain in association of fibrils. Amino acid sequence determinants in self-assembly of insulin chiral amyloid superstructures: role of C-terminus of B-chain in association of fibrils. FEBS Lett. (0014-5793) 2013 Babenko V., Dzwolak W. in vitro In vitro cd circular dichroism CD CD cd predetermined predetermined participant predetermined Babenko V., Dzwolak W. FEBS Lett. (0014-5793) 2013 ins_pig Insulin INS protein protein pig Sus scrofa (Pig) Pig MALWTRLLPLLALLALWAPAPAQAFVNQHLCGSHLVEALYLVCGERGFFYTPKARREAENPQAGAVELGGGLGGLQALALEGPPQKRGIVEQCCTSICSLYQLENYCN 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 CB4491B429858EBE 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 lalba_bovin Alpha-lactalbumin LALBA ALACTA Lactose synthase B protein protein protein bovin Bos taurus (Bovine) Bovine MMSFVSLLLVGILFHATQAEQLTKCEVFRELKDLKGYGGVSLPEWVCTTFHTSGYDTQAIVQNNDSTEYGLFQINNKIWCKDDQNPHSSNICNISCDKFLDDDLTDDIMCVKKILDKVGINYWLAHKALCSEKLDQWLCEKL mint homomint Regulatory subunit of lactose synthase, changes the substrate specificity of galactosyltransferase in the mammary gland making glucose a good acceptor substrate for this enzyme. This enables LS to synthesize lactose, the major carbohydrate component of milk. In other tissues, galactosyltransferase transfers galactose onto the N-acetylglucosamine of the oligosaccharide chains in glycoproteins 810CDB1145901405 ins_bovin Insulin INS protein protein bovin Bos taurus (Bovine) Bovine MALWTRLRPLLALLALWPPPPARAFVNQHLCGSHLVEALYLVCGERGFFYTPKARREVEGPQVGALELAGGPGAGGLEGPPQKRGIVEQCCASVCSLYQLENYCN DIP protein P01317 original sequence version: 131 75307CF78E61C06A lysc_chick Lysozyme C LYZ 1,4-beta-N-acetylmuramidase C Allergen Gal d IV protein protein chick Gallus gallus (Chicken) Chicken MRSLLILVLCFLPLAALGKVFGRCELAAAMKRHGLDNYRGYSLGNWVCAAKFESNFNTQATNRNTDGSTDYGILQINSRWWCNDGRTPGSRNLCNIPCSALLSSDITASVNCAKKIVSDGNGMNAWVAWRNRCKGTDVQAWIRGCRL DIP protein P00698 original sequence version: 158 81E85743FF579468 ins-ins-12 2 BI 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified BI 7 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 mint Figure 2 shows how agitation and presence of NaCl affect formation of insulin amyloid fibrils. All experiments were conducted at low pD (1.9) and at 60°C. The acidic environment promotes dissociation of native hexamers to dimers and monomers while high temperature leads to a partial unfolding of insulin and formation of aggregation-prone intermediate states [35]. Given enough time, these conditions are sufficient to induce amyloid formation. However, at this low pD, the net charge of insulin aggregate is positive (isoelectric point of insulin being 5.5) and fibrils strongly repel one another hence remain singly dispersed as is apparent from the corresponding AFM image in Figure 2A. In order to decrease the repulsive forces, chargeshielding chloride ions may be added to insulin samples. The Debye screening provided by chloride ions decreases the electrostatic barrier and promotes merging of fibrils into clusters visible in AFM (Figure 2B). Hydrodynamic forces are known to facilitate grouping of dispersed particles into larger, often superstructural arrangements [36, 37] hence, are also expected to act against the repulsive electrostatic forces between fibrils even in the absence of NaCl. An AFM image of aggregates formed upon vortexing of acidified insulin at 60°C (Figure 2C) reveals clusters of finely broken fibrils stacked into amorphous clumps. Finally, Figure 2D shows insulin amyloid superstructures formed when both vortex forces and NaCl affect the aggregation process. f2a f2b f2c f2d f4d f4h lyz-lyz-5 2 HEWL 8 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified HEWL 8 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 f4b homomint mint The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment. lalba-lalba-1 2 α-LAC 6 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified α-LAC 6 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 f4c homomint mint The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment. ins-ins-13 2 HI 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified HI 5 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 f4e homomint mint The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment. ins-ins-14 2 PI 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified PI 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 f4f mint The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment. ins-ins-15 1 BI 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified BI 7 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 f2e mint Corresponding FTIR spectra of all the four types of insulin amyloid obtained under different aggregation regimes are shown in Figure 2E. The positions and shapes of the amide I‟ bands are all indicative of parallel - sheet structure while the minor differences should be attributed to variations in inter-β-strand hydrogen bonding and local -sheet twist ins-ins-16 3 BI 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified BI 7 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 mint The most important difference between insulin fibrils obtained under different conditions examined so far is captured by ICD (Figure 2F). The accidental stacking of fibrils in the absence of either NaCl or agitation fails to produce structures with the characteristic strong ICD signals. The chiral superstructures could only be obtained upon vortexing of low-pD insulin samples in the presence of salt and this is reflected by the strong negative peak clearly visible in the corresponding ICD spectrum. f2f f3 ins-ins-17 3 HI 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified HI 5 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 f3 homomint mint For BI, HI, and PI forms of insulin the absolute magnitude of ICD signals is high and comparable. The situation changes dramatically when the amino acid substitutions preventing formation of native dimers are introduced to the precursor (lispro vs. HI Figure 3). The ICD intensity decreases abruptly by at least an order of magnitude, although as noted earlier, its sign remains positive as for PI and HI (inset of Figure 3). The ICD signal for lispro aggregates becomes comparable to the values obtained for HEWL, -LAC, or PLGA fibrils i.e. its origins are conformational (chiral twisting of trapped ThT molecules) rather than superstructural (long-range couplings of helicallyarranged ThT molecules on amyloid scaffold). Similar observations have been described in our previous works for different secondary structures of PLGA, for which both ThT-ICD and far-UV CD peaks had markedly lower intensities [42]. The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment. ins-ins-18 3 PI 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified PI 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 f3 mint For BI, HI, and PI forms of insulin the absolute magnitude of ICD signals is high and comparable. The situation changes dramatically when the amino acid substitutions preventing formation of native dimers are introduced to the precursor (lispro vs. HI Figure 3). The ICD intensity decreases abruptly by at least an order of magnitude, although as noted earlier, its sign remains positive as for PI and HI (inset of Figure 3). The ICD signal for lispro aggregates becomes comparable to the values obtained for HEWL, -LAC, or PLGA fibrils i.e. its origins are conformational (chiral twisting of trapped ThT molecules) rather than superstructural (long-range couplings of helicallyarranged ThT molecules on amyloid scaffold). Similar observations have been described in our previous works for different secondary structures of PLGA, for which both ThT-ICD and far-UV CD peaks had markedly lower intensities [42]. The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment. lyz-lyz-6 3 HEWL 8 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified HEWL 8 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 f3 homomint mint For BI, HI, and PI forms of insulin the absolute magnitude of ICD signals is high and comparable. The situation changes dramatically when the amino acid substitutions preventing formation of native dimers are introduced to the precursor (lispro vs. HI Figure 3). The ICD intensity decreases abruptly by at least an order of magnitude, although as noted earlier, its sign remains positive as for PI and HI (inset of Figure 3). The ICD signal for lispro aggregates becomes comparable to the values obtained for HEWL, -LAC, or PLGA fibrils i.e. its origins are conformational (chiral twisting of trapped ThT molecules) rather than superstructural (long-range couplings of helicallyarranged ThT molecules on amyloid scaffold). Similar observations have been described in our previous works for different secondary structures of PLGA, for which both ThT-ICD and far-UV CD peaks had markedly lower intensities [42]. The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment. lalba-lalba-2 3 α-LAC 6 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified α-LAC 6 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 f3 homomint mint For BI, HI, and PI forms of insulin the absolute magnitude of ICD signals is high and comparable. The situation changes dramatically when the amino acid substitutions preventing formation of native dimers are introduced to the precursor (lispro vs. HI Figure 3). The ICD intensity decreases abruptly by at least an order of magnitude, although as noted earlier, its sign remains positive as for PI and HI (inset of Figure 3). The ICD signal for lispro aggregates becomes comparable to the values obtained for HEWL, -LAC, or PLGA fibrils i.e. its origins are conformational (chiral twisting of trapped ThT molecules) rather than superstructural (long-range couplings of helicallyarranged ThT molecules on amyloid scaffold). Similar observations have been described in our previous works for different secondary structures of PLGA, for which both ThT-ICD and far-UV CD peaks had markedly lower intensities [42]. The corresponding AFM images of different aggregates are presented in Figure 4. Although all amyloid fibrils formed in agitated solutions are strongly associated, aggregates of BI, and HI reveal a higher degree of lateral alignment.