Fibrils Formed in Vitro from α-Synuclein and Two Mutant Forms Linked to Parkinson's Disease are Typical Amyloid

TLDR

Two missense mutations in alpha‑synuclein are linked to rare, early‑onset Parkinson’s disease, which, like idiopathic PD, is marked by Lewy bodies containing fibrillar alpha‑synuclein resembling amyloid plaques of Alzheimer’s disease. The authors produced in vitro fibrils from wild‑type and mutant alpha‑synuclein and demonstrated that they share classic amyloid characteristics—unbranched, wound morphology, Congo red and thioflavin T binding, and antiparallel β‑sheet structure—using atomic force, electron microscopy, FTIR, and CD spectroscopy. The fibrils are protease‑resistant, supporting the view that Parkinson’s is a brain amyloid disease with cytoplasmic amyloid, and a small oligomeric alpha‑synuclein species observed before fibril formation may be the neurotoxic agent.

Abstract

Two missense mutations in the gene encoding alpha-synuclein have been linked to rare, early-onset forms of Parkinson's disease (PD). These forms of PD, as well as the common idiopathic form, are characterized by the presence of cytoplasmic neuronal deposits, called Lewy bodies, in the affected region of the brain. Lewy bodies contain alpha-synuclein in a form that resembles fibrillar Abeta derived from Alzheimer's disease (AD) amyloid plaques. One of the mutant forms of alpha-synuclein (A53T) fibrillizes more rapidly in vitro than does the wild-type protein, suggesting that a correlation may exist between the rate of in vitro fibrillization and/or oligomerization and the progression of PD, analogous to the relationship between Abeta fibrillization in vitro and familial AD. In this paper, fibrils generated in vitro from alpha-synuclein, wild-type and both mutant forms, are shown to possess very similar features that are characteristic of amyloid fibrils, including a wound and predominantly unbranched morphology (demonstrated by atomic force and electron microscopies), distinctive dye-binding properties (Congo red and thioflavin T), and antiparallel beta-sheet structure (Fourier transform infrared spectroscopy and circular dichroism spectroscopy). alpha-Synuclein fibrils are relatively resistant to proteolysis, a property shared by fibrillar Abeta and the disease-associated fibrillar form of the prion protein. These data suggest that PD, like AD, is a brain amyloid disease that, unlike AD, is characterized by cytoplasmic amyloid (Lewy bodies). In addition to amyloid fibrils, a small oligomeric form of alpha-synuclein, which may be analogous to the Abeta protofibril, was observed prior to the appearance of fibrils. This species or a related one, rather than the fibril itself, may be responsible for neuronal death.