Main description:

This detailed book gathers a broad collection of experimental approaches to assist researchers in setting up different methods to investigate protein conformational disorders. Beginning with a section on assays focusing on biophysical approaches to study protein (mis)folding, the volume continues with sections on cellular and proteostasis assays as well as assays for protein folding correction and recovery, combining methods such as thermal shift assays, in silico improvement of protein solubility, and compound screening, an important area of research as it may open avenues for therapeutic strategies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Protein Misfolding Diseases: Methods and Protocols serves as an ideal guide for researchers seeking to advance our knowledge of protein conformational disorders.

Contents:

Part I: Protein Biophysics Assays

1. Biophysical and Spectroscopic Methods for Monitoring Protein Misfolding and Amyloid Aggregation

Joana S. Cristovao, Barbara J. Henriques, and Claudio M. Gomes

2. Ultrasensitive RT-QuIC Seed Amplification Assays for Disease-Associated Tau, -Synuclein, and Prion Aggregates

Eri Saijo, Bradley R. Groveman, Allison Kraus, Michael Metrick, Christina D. Orru, Andrew G. Hughson, and Byron Caughey

3. Vesicle-Based Assays to Study Membrane Interactions of Amyloid Peptides

Ravit Malishev, Sofiya Kolusheva, and Raz Jelinek

4. Differential Scanning Fluorimetry and Hydrogen Deuterium Exchange Mass Spectrometry to Monitor the Conformational Dynamics of NBD1 in Cystic Fibrosis

Naoto Soya, Ariel Roldan, and Gergely L. Lukacs

5. A Multipronged Method for Unveiling Subtle Structural-Functional Defects of Mutant Chaperone Molecules Causing Human Chaperonopathies

Donatella Bulone, Pier Luigi San Biagio, Tatiana Quinones-Ruiz, Manuel Rosario-Alomar, Igor K. Lednev, Frank T. Robb, Everly Conway de Macario, and Alberto J.L. Macario

6. High-Throughput Microplate-Based Fluorescence Assays for Studying Stochastic Aggregation of Superoxide Dismutase-1

Alireza Abdolvahabi, Sanaz Rasouli, Corbin M. Croom, and Devon L. Plewman

7. Methods for Structural Analysis of Amyloid Fibrils in Misfolding Diseases

Devkee M. Vadukul, Youssra K. Al-Hilaly, and Louise C. Serpell

8. Assays for Light Chain Amyloidosis Formation and Cytotoxicity

Luis M. Blancas-Mejia, Pinaki Misra, Christopher J. Dick, Marta Marin Argany, Keely R. Redhage, Shawna A. Cooper, and Marina Ramirez-Alvarado

Part II: Cellular And Proteostasis Assays

9. Monitoring Aggregate Clearance and Formation in Cell-Based Assays

Evelien Eenjes, Young Joo Yang-Klingler, and Ai Yamamoto

10. Monitoring Proteome Stress in Live Cells Using HaloTag-Based Fluorogenic Sensor

Yu Liu, Matthew Fares, and Xin Zhang

11. Quantification of Protein Aggregates Using Bimolecular Fluorescence Complementation

Vibha Prasad and Aaron Voigt

12. Screening Protein Aggregation in Cells Using Fluorescent Labels Coupled to Flow Cytometry

Salvador Ventura and Susanna Navarro

13. Induction of Cu/Zn Superoxide Dismutase (SOD1) Aggregation in Living Cells

Edward Pokrishevsky, Jeremy Nan, and Neil R. Cashman

14. A Cell Model for HSP60 Deficiencies: Modeling Different Levels of Chaperonopathies Leading to Oxidative Stress and Mitochondrial Dysfunction

Cagla Coemert, Paula Fernandez Guerra, and Peter Bross

15. Super-Resolution Fluorescence Imaging of Mutant Huntingtin Aggregation in Cells

Steffen J. Sahl and Willianne I.M. Vonk

Part III: Protein Folding Recovery and Correction Assays

16. Thermal Shift and Stability Assays of Disease-Related Misfolded Proteins Using Differential Scanning Fluorimetry

Tania G. Lucas, Claudio M. Gomes, and Barbara J. Henriques

17. Methods to Screen Compounds against Mutant p53 Misfolding and Aggregation for Cancer Therapeutics

Giulia Diniz da Silva Ferretti, Danielly Ferraz da Costa, Jerson Lima da Silva, and Luciana Pereira Rangel

18. Early Stage Discovery and Validation of Pharmacological Chaperones for the Correction of Protein Misfolding Diseases

Oscar Aubi, Per M. Knappskog, and Aurora Martinez

19. Constructing Kinetically Controlled Denaturation Isotherms of Folded Proteins Using Denaturant-Pulse Chaperonin Binding

Pierce T. O'Neil, Alexandra J. Machen, Jackie A. Thompson, Wei Wang, Quyen Q. Hoang, Michael R. Baldwin, Karen R. Khar, John Karanicolas, and Mark T. Fisher

20. In Vitro Prion Amplification Methodology for Inhibitor Screening

Tuane Cristine R.G. Vieira and Jerson L. Silva

21. SolubiS: Optimizing Protein Solubility by Minimal Point Mutations

Rob van der Kant, Joost van Durme, Frederic Rousseau, and Joost Schymkowitz