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Svanbergsson, A.

Publications and source records attributed to Svanbergsson, A..

2 recordsLinked to original sources

The bacterial amyloids phenol soluble modulins from Staphylococcus aureus catalyze alpha-synuclein aggregation

Aggregated -synuclein (-syn) is the main constituent of Lewy bodies, the main pathological hallmark of Parkinsons disease (PD). Environmental factors are thought to be potential triggers capable of initiating the aggregation of the otherwise monomeric -syn. Braaks seminal work redirected attention to the intestine and recent reports of dysbiosis have highlighted the potential causative role that the microbiome might play in the pathology of PD. Staphylococcus aureus is a bacterium carried by 30-70% of the general population. It has been shown to produce functional amyloids, called Phenol Soluble Modulins (PSMs). Here, we studied the kinetics of -syn aggregation under quiescent conditions in the presence or absence of four different PSM peptides and observed a remarkable shortening of the lag phase in their presence. Whereas pure -syn monomer did not aggregate up to 450 h after initiation of the experiment in neither neutral nor mildly acidic buffer, the addition of different PSM peptides resulted in an almost immediate increase in the Thioflavin T (ThT) fluorescence. Despite similar peptide sequences, the different PSM peptides displayed distinct effects on the kinetics of -syn aggregation. Kinetic analyses of the data suggest that while all four peptides catalyze -syn aggregation, the underlying mechanisms might differ with a model of nucleation and elongation fitting the -syn aggregation induced by PSM2 but not PSM1. The results of immunogold TEM imply that the aggregates are fibrillar and composed of -syn. Addition of the co-aggregated materials to HEK cells expressing the A53T -syn variant fused to GFP was found to catalyze -syn aggregation and phosphorylation in the cells. Our results provide evidence of a potential trigger of synucleinopathies and could have implications for the prevention of the diseases.

biochemistry

FRET-based screening in HEK293T identifies p38 MAPK and PKC inhibition as therapeutic targets for α-synuclein aggregation

Aggregation of -synuclein is associated with neurodegeneration and a hallmark pathology in synucleinopathies. These aggregates are thought to function as prion-like particles where the conformation of misfolded -synuclein determines the induced pathologys traits similar to prion diseases. Still, little is known about the molecular targets facilitating the conformation-specific biological effects, but their identification could form the basis for new therapeutic intervention. High-throughput screening (HTS) of annotated compound libraries could facilitate mechanistic investigation by identifying targets with impact on -synuclein aggregation. To this end, we developed a FRET-based cellular reporter in HEK293T cells, with sensitivity down to 6.5 nM -synuclein seeds. Using this model system, we identified GF109203X, SB202190, and SB203580 as inhibitors capable of preventing induction of - synuclein aggregation via inhibition of p38 MAPK and PKC, respectively. Our findings highlight the value HTS brings to the mechanistic investigation of -synuclein aggregation while simultaneously identifying novel therapeutic compounds.

molecular biology