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Prion Protein

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Contents

Contributors

Preface

Chapter One: Functions of the Prion Protein

1. Introduction

2. The Basics of PrPC

2.1. PrPC From Gene to Function

2.2. PrPC Expression and Localization

3. Lessons From Knockout Models

4. Miscellaneous Roles in CNS Function

4.1. Cognitive and Behavioral Functions

4.2. Neuritogenesis and Neuronal Differentiation

4.3. Balancing Neuroprotection and Neurotoxicity

4.4. Beyond Neurons: PrPC in Glia

4.5. The Metal Connection

4.6. From Neurotransmitter Receptors to Neurotransmitter-Associated Functions

5. Cell Fate and Differentiation

5.1. Stem Cell Self-renewal and Expansion

5.2. A Hub in Developmental Signaling Pathways

5.3. Choice of Cell Fate and Differentiation

6. A Broad Protective Role Against Stress

6.1. PrPC Is Induced Under Stress

6.2. Protection Against Oxidative Stress

6.3. Protection Against Genotoxic Stress

6.4. Protection Against Apoptosis

6.5. An Immune Privilege Molecule?

6.6. Involvement in Proteostasis

7. Cell Adhesion and ECM Contacts

7.1. Multiple Adhesion Partners

7.2. Special Links With Cadherins

7.3. Modulating the ECM Composition

8. From Partners to Cell Signaling

8.1. Cell Signaling: The Unifying Function of PrPC?

8.2. A Great Variety of Partners

8.3. Overview of PrPC-Dependent Signal Transduction

9. Concluding Remarks

References

Chapter Two: Copper- and Zinc-Promoted Interdomain Structure in the Prion Protein: A Mechanism for Autoinhibition of the ...

1. Introduction

1.1. PrPC Structure

1.2. Molecular Features of Cu2+ and Zn2+ Binding

1.3. Metal Ions and PrPC Function

2. The N-Terminal Toxicity Hypothesis

2.1. Internal Deletions of PrPC

2.2. N-Terminal Toxicity Induced by GD Ligands

3. Cu2+ and Zn2+ Promote Interdomain Interaction in cis

4. Disruption of cis Interaction as a Mechanism of Neurotoxicity

5. Concluding Remarks

Acknowledgment

References

Chapter Three: Cell Biology of Prion Protein

1. Expression and Function of Prion Protein

2. PrPC: The Principal Actor in Prion Replication

3. Structure and Biosynthesis of PrPC

4. Trafficking of Cellular Prion Protein

5. Several Topological Forms of PrPC

6. Quality Control of Cellular Prion Protein

7. GPI-Anchor: Structure and Biosynthesis

8. Functions of GPI-Anchored Proteins

9. GPI-Anchor Function

10. GPI-Anchor and Prion

References

Chapter Four: Understanding the Effect of Disease-Related Mutations on Human Prion Protein Structure: Insights From NMR S ...

1. Introduction

2. Structural Features of PrPC and PrPSc

3. Strategy for Resonance Assignment and NMR Structure Determination of Human Prion Protein Variants

4. Structural Features of Human Prion Proteins With Pathological Mutations

4.1. Mildly Acidic pH Conditions

4.2. Physiological pH Conditions

5. Structural Features of Human Prion Protein With Protective E219K Polymorphism

6. Concluding Remarks

Acknowledgment

References

Chapter Five: Structural Modeling of Human Prion Protein´s Point Mutations

1. Introduction

2. Wild-Type Human Prion Protein

2.1. Globular Domain

2.2. N-Terminal Domain

3. Prion Protein Variants

3.1. Globular Domain

3.2. N-Terminal Domain

4. Chimeric Prion Protein

5. Conclusions

References

Chapter Six: Prion Protein and Genetic Susceptibility to Diseases Caused by Its Misfolding

1. Introduction

2. Scrapie, Kuru, and Slow Viruses

3. Prions and PrP

4. Genetic Linkage of Scrapie Incubation Time and the PrP Gene

5. Scrapie Strains

5.1. PrP, Passage History, and Prion Strains

5.2. Species Barrier to Prion Transmission

6. Prnp Knockouts

6.1. PrP Function

7. PRNP and Familial Prion Diseases

7.1. Primary Structure Does Not Predict Infectivity

8. PrP Conformation, Prion Strains, and Susceptibility to Human Prion Diseases

8.1. Protein Conformation Enciphers Heritable Information

8.2. Selection and Evolution of Prion Strains

9. Genes in Addition to the PrP Gene That Influence Susceptibility to PrP Prion Diseases

10. Concluding Remarks: Genetics and the Prospects for PrP Prion Disease Therapeutics

References

Chapter Seven: The Prion Concept and Synthetic Prions

1. The Prion Concept

2. Natural and Recombinant PrP Can Misfold In Vitro

3. Synthetic Prions Are Infectious

4. Synthetic Prions Undergo a Process of Adaptation

5. Alternative Approaches to Generate Synthetic Prions

6. Concluding Remarks

References

Chapter Eight: Gene Targeted Transgenic Mouse Models in Prion Research

1. Introduction

2. The Mouse as a Model of Prion Disease

3. The Influence of Host PrP Expression on Disease Susceptibility

4. PrP Mutations and Disease Transmission

5. Posttranslational Modifications Can Influence Disease Transmission

6. Crossing the Species Barrier and Assessing Zoonotic Potential

7. Modeling Human Prion Disease Transmission

8. Defining Human Prion Disease Strains and Transmissibility Potential

9. Understanding the Function of PrP

10. Mechanisms of Neurodegeneration

11. Conclusion

References

Chapter Nine: Transmission and Replication of Prions

1. Prion Replication

1.1. The “Protein-Only” Hypothesis

1.2. General Aspects of Prion Replication

1.3. Prion Replication Models

1.4. Prion Strains

2. Prion Transmission

2.1. Prion Transmissibility and the Species Barrier

2.2. Polymorphic Variants and the Species Barrier

2.3. Routes of Infection

3. Concluding Remarks

References

Chapter Ten: Immunology of Prion Protein and Prions

1. Introduction

2. Prions Infect Secondary Lymphoid Organs (SLO) Before the CNS

2.1. Prion Replication Upon Follicular Dendritic Cells Within SLO Is Essential to Achieve Neuroinvasion

2.2. Prions Exploit M Cells to Enter Peyer's Patches and Establish Host Infection

3. The Contrasting Roles of Mononuclear Phagocytes

3.1. Conventional Dendritic Cells Propagate Prions Toward and Within SLO

3.2. Macrophages Can Engulf and Destroy Prions

3.3. Distinguishing Between Conventional DC and Macrophages

4. B Cells Disseminate Prions Between SLO

5. Prion Neuroinvasion From SLO Occurs via the Peripheral Nervous System

6. Microglia Provide a Host-Protective Role in the CNS

7. Influence of Aging

8. Influence of Inflammation and Pathogen Coinfection

9. Prospects for Immunotherapy Against Prions

10. Concluding Remarks

References

Chapter Eleven: Phenotypical Variability in Bovine Spongiform Encephalopathy: Epidemiology, Pathogenesis, and Diagnosis o ...

1. Introduction

2. Epidemiology

2.1. Geographical Risk Assessment

2.2. Risk Factors and Effective Measures

3. Pathogenesis of Classical and Atypical BSE

4. Sampling

5. Rapid Tests

6. Histopathological Examination

7. Immunohistochemistry

8. Western Blotting

9. Scrapie-Associated Fibrils

10. In Vitro Amplification Techniques

10.1. Protein Misfolding Cyclic Amplification

10.2. Real-Time Quaking-Induced Conversion

References

Chapter Twelve: Scrapie, CWD, and Transmissible Mink Encephalopathy

1. Scrapie

1.1. History and Epidemiology

1.2. Clinical Signs

1.3. Distribution of Scrapie in Sheep Tissues

1.4. Diagnosis

1.5. Transmission and Pathogenesis

1.6. Zoonotic Potential

2. Chronic Wasting Disease

2.1. History and Epidemiology

2.1.1. Elk

2.1.2. Mule Deer and White-Tailed Deer

2.1.3. Moose and Reindeer

2.2. Clinical Signs

2.3. Distribution of CWD in Cervid Tissues

2.3.1. Histopathology

2.3.2. Pattern of PrPCWD Tissue Accumulation

2.4. Diagnosis

2.5. Transmission and Pathogenesis

2.6. Zoonotic Potential

3. Transmissible Mink Encephalopathy

3.1. History and Epidemiology

3.2. Clinical Signs

3.3. Distribution of TME in Mink Tissues

3.3.1. Histopathology

3.4. Diagnosis

3.5. Transmission and Pathogenesis

3.6. Zoonotic Potential

References

Chapter Thirteen: Infectious and Sporadic Prion Diseases

1. Introduction

2. Sporadic CJD

2.1. Introduction

2.2. Epidemiology

2.3. Cause

2.4. Clinical Features

2.5. Diagnosis

3. Variably Protease Sensitive Prionopathy

4. Iatrogenic CJD

4.1. Introduction

4.2. General Characteristics

4.3. Diagnosis

4.4. Human Growth Hormone (hGH) CJD

4.5. Human Gonadotrophin CJD

4.6. Human Dura Mater CJD

4.7. Other Causes

5. Variant CJD

5.1. Introduction

5.2. Epidemiology

5.3. Cause

5.4. Secondary Transmission

5.5. Genetic Factors

5.6. Clinical Features

5.7. Diagnosis

References

Further Reading

Chapter Fourteen: Neuropathology of Human Prion Diseases

1. Introduction

2. Classification of Human Prion Diseases

2.1. Sporadic CJD

2.2. Variably Protease-Sensitive Prionopathy

2.3. Genetic Prion Diseases

2.4. Iatrogenic CJD

2.5. Variant CJD

2.6. Kuru

2.7. Diagnostic Criteria for Human Prion Diseases

3. The Autopsy in Human Prion Diseases

3.1. When to Suspect a Prion Disease Prior to Autopsy

3.2. Autopsy Considerations

4. Neuropathology

4.1. Macroscopic Pathology and Tissue Sampling

4.2. Brain Biopsies

4.3. Microscopy

4.3.1. Microscopic Pathology

4.3.2. Spongiform Change

4.3.3. Amyloid Plaques

4.3.4. Sporadic CJD

4.3.5. VPSPr

4.3.6. Genetic CJD

4.3.7. GSS

4.3.8. FFI

4.3.9. Insertional, Deletion, and Amber PRNP Mutations

4.3.10. Variant CJD

4.3.11. Iatrogenic CJD

5. Methods for the Detection of PrPSc and PrPres in Tissues

5.1. Immunohistochemistry for PrP

5.2. Paraffin-Embedded Tissue Blot for PrPres

5.3. Western Blot for PrPres

5.4. Recent Laboratory Techniques for PrPSc Detection in Human Tissues

6. Conclusions

Acknowledgments

References

Chapter Fifteen: The Structure of the Infectious Prion Protein and Its Propagation

1. Introduction

2. Infectious Prion Protein Structure

2.1. Spectroscopic Techniques

2.2. Chemical Probes

2.3. Limited Proteolysis

2.4. Diffraction Approaches

2.5. Imaging

2.5.1. Scanning Probe Microscopy

2.5.2. Negative Stain Electron Microscopy

2.5.3. Electron Cryomicroscopy

2.6. NMR Spectroscopy

2.7. Molecular Modeling

3. Propagation of Infectious Prions

4. Future Approaches to Investigate the Structure of Prions

5. Concluding Remarks

Acknowledgments

References

Chapter Sixteen: Protein Misfolding Cyclic Amplification of Infectious Prions

1. Introduction

2. PMCA Applications in Basic Research

3. Applications of PMCA as Supportive Test in Prion Diagnosis

4. PMCA Applications in Drug Screening

5. PMCA Applications to Evaluate Prion Inactivation

6. PMCA and Prion-Like Disorders

References

Chapter Seventeen: Amplified Detection of Prions and Other Amyloids by RT-QuIC in Diagnostics and the Evaluation of Thera ...

1. Introduction

2. Diagnostic Applications of RT-QuIC

2.1. Human CJD Diagnosis Using CSF

2.2. Second-Generation RT-QuIC Assay for CJD Diagnosis Using CSF

2.3. Nasal Brushings as Diagnostic Specimens

2.4. Bovine Spongiform Encephalopathies

2.5. Sheep and Goat Scrapie

2.6. Cervid Chronic Wasting Disease

2.7. Virtually Universal Detection of Prions With a Single Substrate

3. Quantitation of Relative Seed Concentrations

4. Therapeutic Applications of RT-QuIC

5. Evaluating Prion Disinfectants Using RT-QuIC

6. Technical Considerations in RT-QuIC Applications

7. Adaptation of RT-QuIC Platform to Other Protein Folding Diseases

Acknowledgments

References

Chapter Eighteen: Biochemical Characterization of Prions

1. Introduction

2. 1D Analysis of Molecular Strains

2.1. Human Prion Disorders

2.1.1. Sporadic Forms

2.1.2. Genetic Forms

2.1.3. Infectious Forms

2.2. Classical and Atypical BSE Forms

2.3. Scrapie

3. Molecular Analysis of Prion Strains by 2D-PAGE Analysis

3.1. 2D-PAGE Analysis of PrPSc in sCJD Molecular Subtypes

3.2. Molecular Signature of PrPSc in P102L GSS Mutation

3.3. The Influence of GPI Anchor in PrPSc Migration in sCJD

3.4. Cooccurrence of PrPSc Types in sCJD

3.5. Molecular Signatures of PrPSc in BSE Forms

3.6. PrPSc GPI-Anchored and -Anchorless Forms in BSE Forms

3.7. Cooccurrence of PrPSc Types in Scrapie

4. Biochemical Assays to Characterize and Distinguish Prion Strains

4.1. Velocity Sedimentation in Sucrose Step Gradients

4.1.1. Size Aggregates in sCJD

4.1.2. PrP Sedimentation Aggregates in Different Forms of BSE

4.2. Conformational Stability in Increasing Concentration of GdnHCl

4.2.1. Conformational Stability Assay in sCJD

4.2.2. Conformational Stability Assay in BSEs

5. Molecular and Chemicophysical Similarities Between Human and Cattle Transmissible Spongiform Encephalopathies Forms

5.1. Comparison of Human and Cattle Molecular PrPSc by 1D Analysis

5.2. Comparison of Human and Cattle Molecular Signature by 2D Analysis ofPrPSc

6. Conclusions

References

Further Reading

Chapter Nineteen: Omics of Prion Diseases

1. Introduction

2. Genome-Wide Association Studies

2.1. GWAS in Animal Prion Models

2.2. GWAS in Human Prion Diseases

3. Microarray-Based Trancriptomic Studies

3.1. Transcriptomic Studies in Animal Models

3.1.1. Rodents Studies

3.1.2. Ovine Studies

3.1.3. Bovine and Cervid Studies

3.2. Transcriptomic Studies in Primates and Humans

4. Sequencing Approach in Prion Disorders

5. Epigenomic Studies

6. Proteomic Studies

7. Concluding Remarks

References

Chapter Twenty: Therapeutic Approaches to Prion Diseases

1. Introduction

2. Investigational Drugs

3. Preclinical Drug Candidates

3.1. Multitarget Antiprion Small Molecules

3.2. Theranostic Antiprion Small Molecules

4. Conclusions

References

Chapter Twenty-One: Biosafety of Prions

1. Introduction

2. Prion Classification

2.1. Natural Prions

2.2. Synthetically Generated Prions

3. Levels of Infectivity in Different Tissues of Animal and Human TSEs

3.1. Tissue Distribution of PrPSc in Animal TSEs

3.2. Tissue Distribution of PrPSc in Human TSEs

4. Risk Assessment

4.1. Transmissibility of Natural and Synthetic Prions

4.1.1. Natural Transmission of Prions

4.1.2. Experimental Transmission of Natural Prions: The “Species Barrier”

4.1.3. Experimental Transmission of Synthetic Prions

4.2. Infectious Dose

5. Infectious Properties of PMCA and RT-QuIC Reaction Products

5.1. Infectivity of PMCA Reaction Products

5.2. Infectivity of RT-QuIC Reaction Products

6. General Biosafety Recommendations

7. Operating Standard Procedures for Working in BSL-2 or BSL-3 Facilities

7.1. Biosafety Level-2 Laboratories (BSL-2)

7.2. Biosafety Level-3 Laboratories (BSL-3)

8. Biochemical and Histological Analysis

9. Inactivation of Prions and IATA Regulations

9.1. IATA Guidelines for the Transport of Infectious Materials

References

Index

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