Membrane Permeabilization by Oligomeric α-Synuclein: In Search of the Mechanism

Background: The question of how the aggregation of the neuronal protein alpha-synuclein contributes to neuronal toxicity in Parkinson's disease has been the subject of intensive research over the past decade. Recently, attention has shifted from the amyloid fibrils to soluble oligomeric intermediates in the alpha-synuclein aggregation process. These oligomers are hypothesized to be cytotoxic and to permeabilize cellular membranes, possibly by forming pore-like complexes in the bilayer. Although the subject of alpha-synuclein oligomer-membrane interactions has attracted much attention, there is only limited evidence that supports the pore formation by alpha-synuclein oligomers. In addition the existing data are contradictory. Methodology/Principal Findings: Here we have studied the mechanism of lipid bilayer disruption by a well-characterized alpha-synuclein oligomer species in detail using a number of in vitro bilayer systems and assays. Dye efflux from vesicles induced by oligomeric alpha-synuclein was found to be a fast all-or-none process. Individual vesicles swiftly lose their contents but overall vesicle morphology remains unaltered. A newly developed assay based on a dextran-coupled dye showed that non-equilibrium processes dominate the disruption of the vesicles. The membrane is highly permeable to solute influx directly after oligomer addition, after which membrane integrity is partly restored. The permeabilization of the membrane is possibly related to the intrinsic instability of the bilayer. Vesicles composed of negatively charged lipids, which are generally used for measuring alpha-synuclein-lipid interactions, were unstable to protein adsorption in general. Conclusions/Significance: The dye efflux from negatively charged vesicles upon addition of alpha-synuclein has been hypothesized to occur through the formation of oligomeric membrane pores. However, our results show that the dye efflux characteristics are consistent with bilayer defects caused by membrane instability. These data shed new insights into potential mechanisms of toxicity of oligomeric alpha-synuclein species.