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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.

Protein Misfolding Diseases: Methods and Protocols

Part I: Protein Biophysics Assays

 

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

            Joana S. Cristóvao, Bárbara J. Henriques, and Cláudio 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 Cömert, 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

            Tânia G. Lucas, Cláudio M. Gomes, and Bárbara 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