3 research outputs found
Complexation of NAC-Derived Peptide Ligands with the C‑Terminus of α‑Synuclein Accelerates Its Aggregation
Aggregation
of α-synuclein (α-Syn) into neurotoxic
oligomers and amyloid fibrils is suggested to be the pathogenic mechanism
for Parkinson’s disease (PD). Recent studies have indicated
that oligomeric species of α-Syn are more cytotoxic than their
mature fibrillar counterparts, which are responsible for dopaminergic
neuronal cell death in PD. Therefore, the effective therapeutic strategies
for tackling aggregation-associated diseases would be either to prevent
aggregation or to modulate the aggregation process to minimize the
formation of toxic oligomers during aggregation. In this work, we
showed that arginine-substituted α-Syn ligands, based on the
most aggregation-prone sequence of α-Syn, accelerate the protein
aggregation in a concentration-dependent manner. To elucidate the
mechanism by which Arg-substituted peptides could modulate α-Syn
aggregation kinetics, we performed surface plasmon resonance (SPR)
spectroscopy, nuclear magnetic resonance (NMR) studies, and all-atom
molecular dynamics (MD) simulation. The SPR analysis showed a high
binding potency of these peptides with α-Syn but one that was
nonspecific in nature. The two-dimensional NMR studies suggest that
a large stretch within the C-terminus of α-Syn displays a chemical
shift perturbation upon interacting with Arg-substituted peptides,
indicating C-terminal residues of α-Syn might be responsible
for this class of peptide binding. This is further supported by MD
simulation studies in which the Arg-substituted peptide showed the
strongest interaction with the C-terminus of α-Syn. Overall,
our results suggest that the binding of Arg-substituted ligands to
the highly acidic C-terminus of α-Syn leads to reduced charge
density and flexibility, resulting in accelerated aggregation kinetics.
This may be a potentially useful strategy while designing peptides,
which act as α-Syn aggregation modulators
The Familial α‑Synuclein A53E Mutation Enhances Cell Death in Response to Environmental Toxins Due to a Larger Population of Oligomers
Amyloid
formation of α-synuclein (α-Syn) and its familial
mutations are directly linked with Parkinson’s disease (PD)
pathogenesis. Recently, a new familial α-Syn mutation (A53E)
was discovered, associated with an early onset aggressive form of
PD, which delays α-Syn aggregation. When we overexpressed wild-type
(WT) and A53E proteins in cells, showed neither toxicity nor
aggregate formation, suggesting merely overexpression may not recapitulate
the PD phenotype in cell models. We hypothesized that cells expressing
the A53E mutant might possess enhanced susceptibility to PD-associated
toxicants compared to that of the WT. When cells were treated with
PD toxicants (dopamine and rotenone), cells expressing A53E showed
more susceptibility to cell death along with compromised mitochondrial
potential and an increased production of reactive oxygen species.
The higher toxicity of A53E could be due to more oligomers being formed
in cells as confirmed by a dot blot assay using amyloid specific OC
and A11 antibody and using an <i>in vitro</i> aggregation
study. The cellular model presented here suggests that along with
familial mutation, environmental and other cellular factors might
play a crucial role in dictating PD pathogenesis
Comparison of Kinetics, Toxicity, Oligomer Formation, and Membrane Binding Capacity of α‑Synuclein Familial Mutations at the A53 Site, Including the Newly Discovered A53V Mutation
The involvement of
α-synuclein (α-Syn) amyloid formation
in Parkinson’s disease (PD) pathogenesis is supported by the
discovery of α-Syn gene (SNCA) mutations linked with familial
PD, which are known to modulate the oligomerization and aggregation
of α-Syn. Recently, the A53V mutation has been discovered, which
leads to late-onset PD. In this study, we characterized for the first
time the biophysical properties of A53V, including the aggregation
propensities, toxicity of aggregated species, and membrane binding
capability, along with those of all familial mutations at the A53
position. Our data suggest that the A53V mutation accelerates fibrillation
of α-Syn without affecting the overall morphology or cytotoxicity
of fibrils compared to those of the wild-type (WT) protein. The aggregation
propensity for A53 mutants is found to decrease in the following order:
A53T > A53V > WT > A53E. In addition, a time course aggregation
study
reveals that the A53V mutant promotes early oligomerization similar
to the case for the A53T mutation. It promotes the largest amount
of oligomer formation immediately after dissolution, which is cytotoxic.
Although in the presence of membrane-mimicking environments, the A53V
mutation showed an extent of helix induction capacity similar to that
of the WT protein, it exhibited less binding to lipid vesicles. The
nuclear magnetic resonance study revealed unique chemical shift perturbations
caused by the A53V mutation compared to those caused by other mutations
at the A53 site. This study might help to establish the disease-causing
mechanism of A53V in PD pathology