Sequence Determinants for Amyloid Fibrillogenesis of Human alpha-Synuclein |
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Authors: | Zibaee Shahin Jakes Ross Fraser Graham Serpell Louise C Crowther R Anthony Goedert Michel |
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Institution: | 1 MRC Laboratory of Molecular Biology, Cambridge,CB2 0QH, UK 2 School of Life Sciences, University of Sussex, Brighton, BN1 9QG, UK |
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Abstract: | Parkinson's disease (PD) and dementia with Lewy bodies (DLB) are characterized by the presence of filamentous inclusions in nerve cells. These filaments are amyloid fibrils that are made of the protein α-synuclein, which is genetically linked to rare cases of PD and DLB. β-Synuclein, which shares 60% identity with α-synuclein, is not found in the inclusions. Furthermore, while recombinant α-synuclein readily assembles into amyloid fibrils, β-synuclein fails to do so. It has been suggested that this may be due to the lack in β-synuclein of a hydrophobic region that spans residues 73-83 of α-synuclein. Here, fibril assembly of recombinant human α-synuclein, α-synuclein deletion mutants, β-synuclein and β/α-synuclein chimeras was assayed quantitatively by thioflavin T fluorescence and semi-quantitatively by transmission electron microscopy. Deletion of residues 73-83 from α-synuclein did not abolish filament formation. Furthermore, a chimera of β-synuclein with α-synuclein(73-83) inserted was significantly less fibrillogenic than wild-type α-synuclein. These findings, together with results obtained using a number of recombinant synucleins, showed a correlation between fibrillogenesis and mean β-strand propensity, hydrophilicity and charge of the amino acid sequences. The combination of these simple physicochemical properties with a previously described calculation of β-strand contiguity allowed us to design mutations that changed the fibrillogenic propensity of α-synuclein in predictable ways. |
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Keywords: | PD Parkinson's disease DLB dementia with Lewy bodies α-syn α-synuclein β-syn β-synuclein ThT thioflavin T NTTF amino-terminally truncated fragment MβP mean β-strand propensity MHL mean hydrophilicity MNC mean net charge MTC mean total charge β-SC β-strand contiguity |
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