Intermediate filament proteins

Content: Front Cover
 Intermediate Filament Proteins
 Copyright
 Dedication
 Contents
 Contributors
 Preface
 References
 Acknowledgments
 Part I: General Methods to Study IF Proteins
 Chapter One: How to Study Intermediate Filaments in Atomic Detail
 1. Introductory Remarks
 2. Bioinformatics
 3. X-ray Crystallography
 3.1. Design of IF Protein Fragments for Crystallization
 3.2. Protein Expression
 3.3. Purification
 3.4. Crystallization
 3.5. X-ray Data Collection
 3.6. Phasing by Molecular Replacement
 3.7. Experimental Phasing Using Heavy Atoms
 3.8. Structure Refinement and Validation 3.9. Limitations and Pitfalls3.10. Obtained Structures and Impact
 4. Electron Paramagnetic Resonance with Site-Directed Spin Labeling
 4.1. Principle
 4.2. Sample Preparation and Measurements
 4.3. Data Interpretation and Impact
 4.4. Limitations and Outlook
 5. Concluding Remarks
 Acknowledgments
 References
 Chapter Two: Mechanical Properties of Intermediate Filament Proteins
 1. Introduction
 2. Viscoelasticity of Purified IFs In Vitro
 3. IFs and the Mechanical Properties of Cells
 3.1. Type III IF: Vimentin and Desmin
 3.1.1. Knockout Models of Vimentin 3.1.2. Drugs and Proteins Disrupting Vimentin3.1.3. Mutated Desmin to Disrupt Vimentin
 3.1.4. Mutated Desmin to Disrupt Desmin
 3.2. Keratins
 3.3. Neurofilaments
 4. Conclusion
 Acknowledgments
 References
 Chapter Three: Multidimensional Monitoring of Keratin Intermediate Filaments in Cultured Cells and Tissues
 1. Introduction
 2. 3D Imaging of Keratin Intermediate Filaments in Cultured Cells
 2.1. Microscopes
 2.2. Preparation of Imaging Medium
 2.3. Surface Coating
 2.4. Preparation of Cells for Imaging
 2.5. Image Acquisition
 2.6. Keratin Network Normalization 2.7. Measuring Keratin Movement2.8. Keratin Bulk Flow Analysis
 2.9. Keratin Turnover Measured by FRAP
 3. 3D Imaging of Keratin Intermediate Filaments in Murine Preimplantation Embryos
 3.1. Embryo Collection and Cultivation
 3.2. Imaging of Preimplantation Embryos
 3.3. Image Processing and Analysis
 4. Outlook
 Acknowledgments
 References
 Chapter Four: Phospho-Specific Antibody Probes of Intermediate Filament Proteins
 1. Introduction
 2. Production of Site- and Phosphorylation State-Specific Antibodies
 2.1. Designing a Synthetic Phosphopeptide 2.2. Conjugation of a Phosphopeptide to a Carrier Protein3. Characterization of Site- and Phosphorylation State-Specific Antibodies
 3.1. ELISA
 3.2. Immunoblotting
 4. Immunocytochemistry
 4.1. Overview for Immunostaining Methods
 4.2. Vimentin Phosphorylation in Mitosis
 4.3. Vimentin Phosphorylation in Signal Transduction
 5. Other Applications
 6. Conclusions
 Acknowledgments
 References
 Chapter Five: Assays for Posttranslational Modifications of Intermediate Filament Proteins
 1. Introduction
 1.1. Posttranslational Modifications of Intermediate Filament Proteins