Navigating through sequence space to understand Alpha-Synuclein Aggregation

The aggregation of the 140-residue protein a-synuclein into amyloid fibres is associated with neurodegenerative diseases known as synucleinopathies including Parkinson's disease (the second most common neurodegenerative disease worldwide). It has been known for many years that a region within this intrinsically disordered protein known as NAC is necessary and sufficient for amyloid formation. Our, and other groups, however, have shown that regions up- and downstream to NAC control the aggregation of a-synuclein to such an extent that mutations in these regions can speed up or completely inhibit aggregation. These observations open new avenues to the development of effective therapeutics to inhibit a-synuclein aggregation, but to do this it is necessary to understand structurally how the regions flanking NAC control its aggregation properties.

To achieve this goal you will first explore the role that every residue of a-synuclein plays in its aggregation using an exciting and powerful new high throughput screening assay developed in our group (the tripartite b-lactamase assay). Once these 'hotspot residues' have been identified, variants with altered aggregation propensity will be generated and their conformational properties determined using a wide range of cutting-edge biophysical approaches (including NMR, CD, single molecule FRET, MS), alongside measurement of their aggregation properties in vitro and in cells.