Effects of the novel K58N mutation on alpha-synuclein aggregation in cell models

December 14, 2022

Parkinson's disease (PD) is the second most common neurodegenerative disorder. Its pathological hallmarks are the degeneration of dopaminergic neurons in the substantia nigra and the intracellular accumulation of proteinaceous inclusions, Lewy bodies (LBs). Despite many advances in the understanding of PD, the precise mechanisms behind its onset are still poorly understood. The first gene that was discovered to be implicated in PD was SNCA, and this gene encodes for alpha synuclein (aSyn), which is the main constituent of LBs. Many mutations in the SNCA gene have been associated with the onset of PD, from multiplications of the locus to single nucleotide variants. Several point mutations in the aSyn protein have been associated with familial forms of PD. Recently, a new mutation, K58N, has been discovered by our collaborators. Thus, the aim of this project was to study this new mutation in terms of aggregation patterns. With this in mind, we used a well-established aggregation model, and characterized these inclusions with the use of nanobodies, and different types of microscopy techniques to investigate possible differences in the aggregation of wild type (WT) vs. K58N aSyn proteins. We discovered that this new mutation is less prone to aggregate when compared to the WT, even though algorithms that predict aggregation suggested otherwise. We also discovered that a novel nanobody, NB40, detects a higher quantity of aggregates per cell in both WT and K58N-mutant aSyn, when compared to the other nanobodies and antibodies used.

Authors in the community:

• 

  G

  Gonçalo Miguel Patrício Mestre

  ist189749

Supervisors of this institution:

• 

  M

  Maria Margarida Fonseca Rodrigues Diogo

  ist24804

industrial-biotechnology - Industrial Biotechnology

eng - English

Instituto Superior Técnico