Probing the Influence of Single-Site Mutations in the Central Cross-ß Region of Amyloid ß (1-40) Peptides.
Biomolecules
; 11(12)2021 12 09.
Article
em En
| MEDLINE
| ID: mdl-34944492
ABSTRACT
Amyloid ß (Aß) is a peptide known to form amyloid fibrils in the brain of patients suffering from Alzheimer's disease. A complete mechanistic understanding how Aß peptides form neurotoxic assemblies and how they kill neurons has not yet been achieved. Previous analysis of various Aß40 mutants could reveal the significant importance of the hydrophobic contact between the residues Phe19 and Leu34 for cell toxicity. For some mutations at Phe19, toxicity was completely abolished. In the current study, we assessed if perturbations introduced by mutations in the direct proximity of the Phe19/Leu34 contact would have similar relevance for the fibrillation kinetics, structure, dynamics and toxicity of the Aß assemblies. To this end, we rationally modified positions Phe20 or Gly33. A small library of Aß40 peptides with Phe20 mutated to Lys, Tyr or the non-proteinogenic cyclohexylalanine (Cha) or Gly33 mutated to Ala was synthesized. We used electron microscopy, circular dichroism, X-ray diffraction, solid-state NMR spectroscopy, ThT fluorescence and MTT cell toxicity assays to comprehensively investigate the physicochemical properties of the Aß fibrils formed by the modified peptides as well as toxicity to a neuronal cell line. Single mutations of either Phe20 or Gly33 led to relatively drastic alterations in the Aß fibrillation kinetics but left the global, as well as the local structure, of the fibrils largely unchanged. Furthermore, the introduced perturbations caused a severe decrease or loss of cell toxicity compared to wildtype Aß40. We suggest that perturbations at position Phe20 and Gly33 affect the fibrillation pathway of Aß40 and, thereby, influence the especially toxic oligomeric species manifesting so that the region around the Phe19/Leu34 hydrophobic contact provides a promising site for the design of small molecules interfering with the Aß fibrillation pathway.
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MEDLINE
Assunto principal:
Peptídeos beta-Amiloides
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Mutação
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article