CONICET | Buscador de Institutos y Recursos Humanos

Alpha-synuclein (a-synuclein) is a major component of intraneuronal protein aggregates constituting a distinctive feature of Parkinsons disease. To date, fluorescence imaging of dynamic processes leading to such amyloid deposits in living cells has not been feasible. To address this need, we generated a recombinant a-synuclein (a-synuclein-C4) bearing a tetracysteine target for fluorogenic biarsenical compounds. The biophysical, biochemical and aggregation properties of a-synuclein-C4 matched those of the wild-type protein in vitro and in living cells. We observed aggregation of a-synuclein-C4 transfected or microinjected into cells, particularly under oxidative stress conditions. Förster resonance energy transfer (FRET) between FlAsH and ReAsH confirmed the close association of fibrillized a-synuclein-C4 molecules. a-synuclein-C4 offers the means for directly probing amyloid formation and interactions of a-synuclein with other proteins in living cells, the response to cellular stress, and screening drugs for Parkinsons disease.