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Biophysical and Spectroscopic Methods for Monitoring Protein Misfolding and Amyloid Aggregation

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Protein Misfolding Diseases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1873))

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).

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Abbreviations

CD:

Circular dichroism

FTIR:

Fourier-transformed infrared spectroscopy

DLS:

Dynamic light scattering

SOD1:

Superoxide dismutase 1

1,8-ANS:

8-Anilinonaphthalene-1-sulfonic acid

ThT:

Thioflavin T

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Acknowledgments

This work was partially supported by the Fundação para a Ciência e Tecnologia (FCT/MCTES, Portugal) through fellowships to J.S.C. (SFRH/BD/101171/2014) and B.J.H. (SFRH/BPD/74475/2010), and grant PTDC/BBB-BQB/5366/2014 (to B.J.H.) and PTDC/NEU-NMC/2138/2014 (to C.M.G.). The Gomes laboratory is partly supported by grant UID/MULTI/04046/2013 from FCT/MCTES/PIDDAC (to BioISI). Bial Foundation is acknowledged through grant PT/FB/BL-2014-343 (to CMG). Joana S. Cristóvão and Bárbara J. Henriques contributed equally to this work.

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Author notes

  1. Joana S. Cristóvão and Bárbara J. Henriques contributed equally to this work.

Authors and Affiliations

  1. BioISI – Biosystems and Integrative Sciences Institute, Faculty of Sciences University of Lisbon, Lisbon, Portugal

    Joana S. Cristóvão, Bárbara J. Henriques & Cláudio M. Gomes

  2. Department of Chemistry and Biochemistry, Faculty of Sciences University of Lisbon, Lisbon, Portugal

    Joana S. Cristóvão, Bárbara J. Henriques & Cláudio M. Gomes

Authors
  1. Joana S. Cristóvão
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  2. Bárbara J. Henriques
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  3. Cláudio M. Gomes
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Corresponding author

Correspondence to Cláudio M. Gomes .

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Editors and Affiliations

  1. BioISI – Biosystems and Integrative Sciences Institute, Faculty of Sciences University of Lisbon, Lisbon, Portugal

    Cláudio M. Gomes

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Cristóvão, J.S., Henriques, B.J., Gomes, C.M. (2019). Biophysical and Spectroscopic Methods for Monitoring Protein Misfolding and Amyloid Aggregation. In: Gomes, C. (eds) Protein Misfolding Diseases. Methods in Molecular Biology, vol 1873. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8820-4_1

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  • DOI: https://doi.org/10.1007/978-1-4939-8820-4_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8819-8

  • Online ISBN: 978-1-4939-8820-4

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