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 Identification of an amyloid fibril forming peptide comprising residues 46-59 of apolipoprotein A-I. Identification of an amyloid fibril forming peptide comprising residues 46-59 of apolipoprotein A-I. FEBS Lett. (0014-5793) 2012 Wong YQ., Binger KJ., Howlett GJ., Griffin MD. suggested by lperfetto in vitro In vitro fluorescence fluorescence technology fluorescence peptide synthesis peptide synthesis Wong YQ., Binger KJ., Howlett GJ., Griffin MD. suggested by lperfetto FEBS Lett. (0014-5793) 2012 Identification of an amyloid fibril forming peptide comprising residues 46-59 of apolipoprotein A-I. Identification of an amyloid fibril forming peptide comprising residues 46-59 of apolipoprotein A-I. FEBS Lett. (0014-5793) 2012 Wong YQ., Binger KJ., Howlett GJ., Griffin MD. suggested by lperfetto in vitro In vitro x-ray fiber diffrac x-ray fiber diffraction X-ray X-ray x-ray fiber diffrac peptide synthesis peptide synthesis Wong YQ., Binger KJ., Howlett GJ., Griffin MD. suggested by lperfetto FEBS Lett. (0014-5793) 2012 Identification of an amyloid fibril forming peptide comprising residues 46-59 of apolipoprotein A-I. Identification of an amyloid fibril forming peptide comprising residues 46-59 of apolipoprotein A-I. FEBS Lett. (0014-5793) 2012 Wong YQ., Binger KJ., Howlett GJ., Griffin MD. suggested by lperfetto in vitro In vitro tem transmission electron microscopy tem predetermined predetermined participant predetermined Wong YQ., Binger KJ., Howlett GJ., Griffin MD. suggested by lperfetto FEBS Lett. (0014-5793) 2012 apoa1_human Apolipoprotein A-I APOA1 Apolipoprotein A1 protein protein human Homo sapiens Human MKAAVLTLAVLFLTGSQARHFWQQDEPPQSPWDRVKDLATVYVDVLKDSGRDYVSQFEGSALGKQLNLKLLDNWDSVTSTFSKLREQLGPVTQEFWDNLEKETEGLRQEMSKDLEEVKAKVQPYLDDFQKKWQEEMELYRQKVEPLRAELQEGARQKLHELQEKLSPLGEEMRDRARAHVDALRTHLAPYSDELRQRLAARLEALKENGGARLAEYHAKATEHLSTLSEKAKPALEDLRQGLLPVLESFKVSFLSALEEYTKKLNTQ DIP protein P02647 original sequence version: 116 DIP protein P02647 original sequence version: 125 DIP protein P02647 original sequence version: 147 DIP protein P02647 original sequence version: 162 1A28B8366E620310 apoa1-apoa1 3 apoA-I 4 ms participant Identification by mass spectrometry ms participant predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain apoA-I 4 ms participant Identification by mass spectrometry ms participant predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f1 homomint mint The aggregates were pelleted by ultracentrifugation and carefully separated from the supernatant. Inspection of the resuspended pellet fraction by transmission electron microscopy (TEM) revealed that this was made up of well defined, regular fibrillar structures of around 20 nm wide and up to several microns in length (Figure 1). To identify the peptide, or peptides, comprising these fibrils, the pellet fraction from the trypsin digest was subjected to reversed phase HPLC, followed by mass spectrometry. HPLC analysis showed a single major eluting peak (Figure S1). Mass spectrometry of this fraction provided a single mass of 1612.8 Da, corresponding to the mass of the tryptic peptide comprising residues 46-59 of apoA-I (apoA-I46-59; LLDNWDSVTSTFSK). Together these results show that the observed fibrils are composed of a single specific peptide. apoa1-apoa1-1 3 apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2b homomint mint TEM of the aggregated apoA-I46-59 peptide sample revealed ordered, fibrillar structures (Figure 2B) similar to those found in the crude tryptic digest apoa1-apoa1-2 3 apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction sf4 homomint mint Truncated apoA-I46-59 peptides show altered capacity for fibril formation. The ability of apoA-I49-59 to form fibrils was confirmed by TEM, which revealed ordered amyloid-like fibrils (Figure S4). apoa1-apoa1-3 1 apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2a homomint mint The time course of development of ThT fluorescence in this sample was characterised by a short lag-phase of around 15 minutes followed by a sharp increase in fluorescence, with aggregation complete after around 4 hours (Figure 2A). apoa1-apoa1-4 1 apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction sf2 homomint mint Tryptophan fluorescence emission data collected for apoA-I46-59 before and after incubation were consistent with aggregation of the peptide (Figure S2) apoa1-apoa1-5 2 apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain apoA-I 4 peptide synthesis peptide synthesis predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2c homomint mint Amyloid fibrils contain a common structure in which two or more large -sheets are formed from repeating -strands, and this structural configuration produces a signature cross- X-ray fibre diffraction pattern [16]. To probe the molecular structure of the apoA-I46-59 fibrils, X-ray diffraction data were collected from samples of partially aligned fibrils using the MX-1 beam-line of the Australian Synchrotron. The diffraction images (Figure 2C) revealed a classical cross- diffraction pattern with orthogonal reflections. The sharp meridional peak at a spacing of 4.75 Å indicates the distance between hydrogen bonded -strands in the direction of the fibril long-axis.