Applications of Chimeric Genes and Hybrid Proteins, Part B: Cell Biology and Physiology (eBook)

Cover 1
Table of Contents 6
Contributors to Volume 327 12
Preface 18
Volumes in Series 20
Section I: Epitope Tags for Immunodetection 39
Chapter 1. Epitope Tagging: General Method for Tracking Recombinant Proteins 41
Section II: Markers for Cytology, Analysis of Protein Trafficking, and Lineage Tracing 55
Chapter 2. Alkaline Phosphatase Fusions of Ligands or Receptors as in Situ Probes for Staining of Cells, Tissues, and Embryos 57
Chapter 3. Chimeric Molecules Employing Horseradish Peroxidase as Reporter Enzyme for Protein Localization in the Electron Microscope 73
Chapter 4. Biochemical Analyses of Trafficking with Horseradish Peroxidase-Tagged Chimeras 83
Chapter 5. Visualizing Protein Dynamics in Yeast with Green Fluorescent Protein 99
Chapter 6. Kinetic Analysis of Intracellular Trafficking in Single Living Cells with Vesicular Stomatitis Virus Protein G–Green Fluorescent Protein Hybrids 107
Chapter 7. Dual Color Detection of Cyan and Yellow Derivatives of Green Fluorescent Protein Using Conventional Fluorescence Microscopy and 35-mm Photography 127
Chapter 8. Invertase Fusion Proteins for Analysis of Protein Trafficking in Yeast 133
Chapter 9. Introduction of Kex2 Cleavage Sites in Fusion Proteins for Monitoring Localization and Transport in Yeast Secretory Pathway 145
Chapter 10. Lineage Analysis with Retroviral Vectors 156
Chapter 11. Use of Pseudotyped Retroviruses in Zebrafish as Genetic Tags 183
Section III: Tools for Analysis of Membrane Proteins 201
Chapter 12. A Gene Fusion Method for Assaying Interactions of Protein Transmembrane Segments in Vivo 203
Chapter 13. Using SUC2–HIS4C Reporter Domain to Study Topology of Membrane Proteins in Saccharomyces cerevisiae 213
Chapter 14. Detecting Interactions between Membrane Proteins in Vivo Using Chimeras 228
Chapter 15. Alkaline Phosphatase Fusion Proteins for Molecular Characterization and Cloning of Receptors and Their Ligands 236
Chapter 16. Surface Chimeric Receptors as Tools in Study of Lymphocyte Activation 248
Chapter 17. Use of Chimeric Receptor Molecules to Dissect Signal Transduction Mechanisms 266
Chapter 18. Fusion Protein Toxins Based on Diphtheria Toxin: Selective Targeting of Growth Factor Receptors of Eukaryotic Cells 277
Chapter 19. Green Fluorescent Protein-Based Sensors for Detecting Signal Transduction and Monitoring Ion Channel Function 287
Chapter 20. Metabolic Labeling of Glycoproteins with Chemical Tags through Unnatural Sialic Acid Biosynthesis 298
Section IV: Signals for Addressing Proteins to Specific Subcellular Compartments 315
Chapter 21. Using Sorting Signals to Retain Proteins in Endoplasmic Reticulum 317
Chapter 22. Directing Proteins to Nucleus by Fusion to Nuclear Localization Signal Tags 321
Chapter 23. Identification, Analysis, and Use of Nuclear Export Signals in Saccharomyces cerevisiae 335
Chapter 24. Directing Proteins to Mitochondria by Fusion to Mitochondrial Targeting Signals 343
Chapter 25. Targeting Proteins to Plasma Membrane and Membrane Microdomains by N-Terminal Myristoylation and Palmitoylation 355
Chapter 26. Analysis of Function and Regulation of Proteins That Mediate Signal Transduction by Use of Lipid-Modified Plasma Membrane-Targeting Sequences 369
Chapter 27. Glycerolphosphoinositide Anchors for Membrane-Tethering Proteins 389
Chapter 28. Fusions to Members of Fibroblast Growth Factor Gene Family to Study Nuclear Translocation and Nonclassic Exocytosis 407
Section V: Application of Chimeras in Monitoring and Manipulating Cell Physiology 421
Chapter 29. Posttranslational Regulation of Proteins by Fusions to Steroid-Binding Domains 423
Chapter 30. Tet Repressor-Based System for Regulated Gene Expression in Eukaryotic Cells: Principles and Advances 439
Chapter 31. Coumermycin-Induced Dimerization of GyrB-Containing Fusion Proteins 459
Chapter 32. Use of Glutathione S-Transferase and Break Point Cluster Region Protein as Artificial Dimerization Domains to Activate Tyrosine Kinases 467
Chapter 33. Recombinant Aequorin as Tool for Monitoring Calcium Concentration in Subcellular Compartments 478
Chapter 34. Recombinant Aequorin as Reporter of Changes in Intracellular Calcium in Mammalian Cells 494
Chapter 35. Monitoring Protein Conformations and Interactions by Fluorescence Resonance Energy Transfer between Mutants of Green Fluorescent Protein 510
Chapter 36. Studies of Signal Transduction Events Using Chimeras to Green Fluorescent Protein 538
Chapter 37. Use of Fusions to Green Fluorescent Protein in the Detection of Apoptosis 551
Chapter 38. Synapto-pHluorins: Chimeras between pH-Sensitive Mutants of Green Fluorescent Protein and Synaptic Vesicle Membrane Proteins as Reporters of Neurotransmitter Release 560
Chapter 39. Studying Organelle Physiology with Fusion Protein-Targeted Avidin and Fluorescent Biotin Conjugates 584
Chapter 40. Fluorescent Labeling of Recombinant Proteins in Living Cells with FlAsH 603
Chapter 41. Ubiquitin Fusion Technique and Its Descendants 616
Chapter 42. Use of Phosphorylation Site Tags in Proteins 632
Author Index 653
Subject Index 695