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In-cell NMR monitoring of alpha-Synuclein aggregation in neuronal cells

Periodic Reporting for period 3 - NeuroInCellNMR (In-cell NMR monitoring of alpha-Synuclein aggregation in neuronal cells)

Reporting period: 2018-10-01 to 2020-07-31

The goal of the NeuroInCellNMR project is to derive atomic-resolution information about the structural states of human alpha-synuclein in the course of amyloid formation in neuronal and non-neuronal cells. Intracellular aggregation of alpha-synuclein is causally linked to Parkinson’s disease, the second most common neurodegenerative disorder in humans. Overall objectives of our research are to delineate novel insights into the conformational rearrangements that initiate and propagate alpha-synuclein aggregation in the course of Parkinson’s disease.
So far, we delineated a first full-scale, atomic-resolution description of intracellular alpha-synuclein structures and dynamics in five different mammalian cell types under physiological conditions (Nature, 2016). We further analyzed the capacity of healthy mammalian cells to repair oxidative-damaged alpha-synuclein, which revealed a site-selective and stepwise process executed by cellular enzymes that collectively failed to target C-terminal modification sites, but efficiently repaired N-terminal damaged residues (Nat Commun, 2016). We developed a novel procedure to determine in-cell protein structures from simple 2D NMR experiments (J Phys Chem Lett, 2016) and applied this approach to resolve the stepwise phosphorylation behavior of a related, disordered protein region (Science, 2016). We further collaborated on the development of a new imaging approach i.e. interferometric scattering microscopy (iSCAT), to directly monitor alpha-synuclein aggregation at membrane interfaces with single molecule resolution (Science, 2018). In parallel, we identified a novel biological activity of human alpha-synculein regarding its ability to interact with endogenous cellular membranes, which serves as the basis for ongoing experiments addressing its aggregation propensity at these membranes.
At this point, we are making good progress in deciphering previously unknown biological scenarios that may precede the intracellular aggregation of alpha-synuclein. We are confident that these insights will allow us to design better strategies to inhibit alpha-synuclein aggregation and the onset of Parkinson’s disease.