Related Experiment Video
Updated: Jun 5, 2025
11:12
Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells
Published on: October 4, 2017
7.2K
Insight Into Factors Influencing the Aggregation Process in Wild-Type and P66R Mutant SOD1: Computational
Roghayeh Farrokhzad1, Bagher Seyedalipour1, Payam Baziyar1
1Department of Molecular and Cell Biology, Faculty of Basic Science, University of Mazandaran, Babolsar, Iran.
Proteins
|December 6, 2024
Summary
Metal dysregulation and mutations in superoxide dismutase (SOD1) trigger protein misfolding and aggregation in amyotrophic lateral sclerosis (ALS). Understanding these mechanisms is key to developing new therapeutic strategies.
Area of Science:
- Biochemistry
- Neuroscience
- Structural Biology
Background:
- Metal ion homeostasis disturbances are linked to amyotrophic lateral sclerosis (ALS).
- The precise role of metalation in superoxide dismutase (SOD1) misfolding and aggregation remains unclear.
Purpose of the Study:
- To investigate the impact of metalation status (apo/holo forms) on the structural and aggregation properties of wild-type (WT) SOD1 and the P66R mutant.
- To correlate physicochemical modifications with protein aggregation at a molecular level.
Main Methods:
- Computational (molecular dynamics simulations) and experimental techniques (FTIR, TEM, fluorescence spectroscopy, GdnHCl denaturation).
- Characterization of WT-SOD1 and P66R mutant in both metal-bound (holo) and metal-deficient (apo) states.
- Assessment of enzymatic activity, flexibility, stability, hydrophobicity, and aggregate formation.
Main Results:
- Apo-SOD1 variants exhibited increased flexibility, altered stability, and hydrophobicity compared to holo-forms.
- Decreased enzymatic activity was observed in P66R mutants compared to WT-SOD1.
- Apo-forms showed increased aggregation propensity, forming β-sheet structures and amyloid aggregates, driven by metal deficit, mutation, and disulfide bond reduction.
- These factors destabilize SOD1, promoting aggregation.
Conclusions:
- Metal deficit, mutations (e.g., P66R), and disulfide bond reduction are critical initiators of SOD1 misfolding and aggregation.
- Disruption of the dimer-monomer equilibrium and reduced thermodynamic stability facilitate toxic aggregate formation.
- Findings provide insights into disease mechanisms and suggest therapeutic targets for protein aggregation disorders like ALS.
