Different Effects of α‑Synuclein Mutants on Lipid Binding and Aggregation Detected by Single Molecule Fluorescence Spectroscopy and ThT Fluorescence-Based Measurements

• Published in: ACS chemical neuroscience Vol. 10; no. 3; pp. 1649 - 1659
• Main Authors: Ruf, Viktoria C, Nübling, Georg S, Willikens, Sophia, Shi, Song, Schmidt, Felix, Levin, Johannes, Bötzel, Kai, Kamp, Frits, Giese, Armin
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 20-03-2019
• Subjects: alpha-Synuclein - metabolism; Humans; Lipids - chemistry; Mutation - genetics; Parkinson Disease - metabolism; Point Mutation - genetics; Protein Aggregates; Protein Aggregation, Pathological - metabolism; Spectrometry, Fluorescence - methods; synucleinopathy; α-Synuclein; protein−lipid interaction; mutant; protein aggregation; Parkinson’s disease
• ISSN: 1948-7193; 1948-7193
• DOI: 10.1021/acschemneuro.8b00579

Six α-synuclein (aSyn) point mutations are currently known to be associated with familial parkinsonism: A30P, E46K, H50Q, G51D, A53E, and A53T. We performed a comprehensive in vitro analysis to study the impact of all aSyn mutations on lipid binding and aggregation behavior. Markedly reduced lipid binding of A30P, moderately attenuated binding of G51D, and only very slightly reduced binding for the other mutants were observed. A30P was particularly prone to form metal ion induced oligomers, whereas A53T exhibited only weak tendencies to form oligomers. In turn, fibril formation occurred rapidly in H50Q, G51D, and A53T, but only slowly in A30P, suggesting mutants prone to form oligomers tend to form fibrils to a lesser extent. This was supported by the observation that fibril formation of wild type aSyn, A30P, and A53T was impaired in the presence of ferric iron. Additionally, we found the aggregation kinetics of mixtures of A30P or A53T and wt aSyn to be determined by the faster aggregating aSyn variant. Our results implicate differential mechanisms playing a role in aSyn pathology on the molecular level. This might contribute to a better understanding of Parkinson’s disease pathogenesis and provide potential links to develop prevention strategies and disease-modifying therapy.