1. Biophysical and Spectroscopic Methods for Monitoring Protein Misfolding and Amyloid Aggregation.
- Author
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Cristóvão JS, Henriques BJ, and Gomes CM
- Subjects
- Amyloidogenic Proteins genetics, Amyloidogenic Proteins metabolism, Amyotrophic Lateral Sclerosis genetics, Amyotrophic Lateral Sclerosis metabolism, Amyotrophic Lateral Sclerosis physiopathology, Anilino Naphthalenesulfonates chemistry, Benzothiazoles chemistry, Fluorescent Dyes chemistry, Gene Expression, Humans, Models, Molecular, Protein Aggregation, Pathological genetics, Protein Aggregation, Pathological metabolism, Protein Aggregation, Pathological physiopathology, Protein Conformation, Protein Folding, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Superoxide Dismutase-1 genetics, Superoxide Dismutase-1 metabolism, Amyloidogenic Proteins chemistry, Circular Dichroism methods, Dynamic Light Scattering methods, Spectrometry, Fluorescence methods, Spectroscopy, Fourier Transform Infrared methods, Superoxide Dismutase-1 chemistry
- Abstract
Proteins exhibit a remarkable structural plasticity and may undergo conformational changes resulting in protein misfolding both in a biological context and upon perturbing physiopathological conditions. Such nonfunctional protein conformers, including misfolded states and aggregates, are often associated to protein folding diseases. Understanding the biology of protein folding diseases thus requires tools that allow the structural characterization of nonnative conformations of proteins and their interconversions. Here we present detailed procedures to monitor protein conformational changes and aggregation based on spectroscopic and biophysical methods that include circular dichroism, ATR-Fourier-transformed infrared spectroscopy, fluorescence spectroscopy and dynamic light scattering. To illustrate the application of these methods we report to our previous studies on misfolding, aggregation and amyloid fibril formation by superoxide dismutase 1 (SOD1), a protein whose toxic deposition is implicated in the neurodegenerative disease amyotrophic lateral sclerosis (ALS).
- Published
- 2019
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