
Endosome Signaling Part A
Description
Key Features
- Continues the legacy of this premier serial with quality chapters authored by leaders in the field
- Covers endosome signaling
- Contains chapters on such topics as measurement of biological effects of endosomal proteolysis of internalized insulin and multi-vesicular endosome biogenesis.
Readership
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists
Table of Contents
Contributors
Preface
Section I: Compartments
Chapter One. Monitoring Phosphatidylinositol 3-Phosphate in Multivesicular Endosome Biogenesis
Abstract
1 Introduction
2 Localization of PtdIns3P and EGFR
3 Correlative Light and Electron Microscopy of MVEs
4 Concluding Remarks
Acknowledgments
References
Chapter Two. Methods to Discriminate the Distribution of Acidic Glycohydrolases Between the Endosomal–Lysosomal Systems and the Plasma Membrane
Abstract
Abbreviations
1 Introduction
2 Purification of Lipid Microdomains from Cell Membranes and Glycohydrolases Activity Determination
3 Discrimination of Cell Surface Lipid Microdomain-Associated Glycohydrolases from the Intracellular Counterparts
4 Immunology Capture of Lipid Microdomains Containing Glycohydrolases
5 In Vivo Assay of Cell Surface Glycohydrolases
6 Fluorescence Microscopy Analysis of Hex Intracellular Trafficking
7 Summary
Acknowledgment
References
Chapter Three. Visualizing of Signaling Proteins on Endosomes Utilizing Knockdown and Reconstitution Approach
Abstract
1 Introduction
2 Description of Methods
3 Conclusions
Acknowledgments
References
Chapter Four. Virus-Induced Signaling Influences Endosome Trafficking, Virus Entry, and Replication
1 Introduction
2 Isolation of Fibroblasts from Wild-Type or MyD88−/− Mice and Purification of Virus
3 Screening Assays for the Determination of Principal Host Kinases Involved During Virus Replication
4 Virus–Host Cell Receptor Interactions and Kinase Activation
5 Detection of Virus-Induced Cell Signaling and Virus Entry into Endosomes Using Immunofluorescence Confocal Microscopy
6 Summary
References
Chapter Five. Methods to Evaluate Zinc Transport into and out of the Secretory and Endosomal–Lysosomal Compartments in DT40 Cells
Abstract
1 Introduction
2 Intracellular Zinc Transporters Localized to the Secretory and Endosomal–Lysosomal Compartments Play Crucial Roles
3 Establishment of DT40 Cells Deficient in Zinc Transporters Genes
4 Experimental Procedures Used in Studies of DT40 Cells Deficient in Zinc Transport
5 Functional Analysis of Zinc Mobilization into or out of the Secretory and Endosomal–Lysosomal Compartments
6 Concluding Remarks
Acknowledgment
References
Chapter Six. Interactions Between Endosomal Maturation and Autophagy: Analysis of ESCRT Machinery During Caenorhabditis elegans Development
Abstract
1 Introduction
2 Strains and Reagents
3 Fluorescent-Tagged Protein Construction and Transgenesis
4 Analysis of Developmental Phenotypes in ESCRT Mutants
5 Analysis of Vesicular Compartments
6 Analysis of Autophagy
7 Methods to Visualize Amphisome, the Fusion Organelle Between Endosomes and Autophagosomes
8 Conclusions
Acknowledgments
References
Chapter Seven. Assessment of Cation Trapping by Cellular Acidic Compartments
Abstract
1 Introduction
2 Quinacrine Uptake by Cells
3 Macroautophagic Accumulation in Cells That Have Accumulated Cations
4 Summary
Acknowledgments
References
Chapter Eight. Signaling Initiated by the Secretory Compartment
Abstract
1 Introduction
2 ER-to-Golgi Traffic-Synchronization Protocols
3 Read-Outs for Traffic-Generated Signaling
4 KDELR Signaling
5 The KDELR Transduction Machinery
6 Conclusions
Acknowledgments
References
Chapter Nine. Image-Based and Biochemical Assays to Investigate Endosomal Protein Sorting
Abstract
Abbreviations
1 Introduction
2 Antibody-Uptake Assays
3 Detailed Characterization of Endosomes
4 Endosome Recruitment and/or Association
5 Summary
Acknowledgments
References
Section II: Transport and Transfer
Chapter Ten. Cytokines, Polarity Proteins, and Endosomal Protein Trafficking and Signaling—The Sertoli Cell Blood–Testis Barrier System In Vitro as a Study Model
Abstract
1 Introduction
2 Endocytosis Assay
3 Materials
4 Buffers
5 Methods
6 Cell Staining to Assess Endocytosis
7 Results
8 Summary
Acknowledgments
References
Chapter Eleven. Methods of Analysis of the Membrane Trafficking Pathway from Recycling Endosomes to Lysosomes
Abstract
1 Introduction
2 Degradation of TfR in Lysosomes
3 Screening Methods for Rab Proteins Involved in Lysosomal Degradation of TfR
4 Effect of Rab12 Knockdown on an EGFR Endocytic Pathway and a Tf Recycling Pathway
5 Concluding Remarks
Acknowledgments
References
Chapter Twelve. Measurement of Intercellular Transfer to Signaling Endosomes
Abstract
1 Introduction
2 Labeling of Transferred Signals
3 Measuring ICT Using Fluorescence Techniques
4 Signaling Endosomes
5 Experimental Example: ICT to SARA-Positive Signaling Endosomes
6 Summary
References
Chapter Thirteen. Liposome-Based Assays to Study Membrane-Associated Protein Networks
Abstract
1 Introduction
2 Isolation of Core Machineries Required for Carrier Biogenesis on Synthetic Membranes
3 Identification of Core Machineries by Mass Spectrometry-Based, Label-Free Quantitative Proteomics
4 Visualization of Protein Dynamics on Giant Unilamellar Vesicles by Fluorescence Microscopy
5 Summary
Acknowledgments
References
Chapter Fourteen. Mouse Models of PI(3,5)P2 Deficiency with Impaired Lysosome Function
Abstract
1 Introduction
2 Design of Mouse Models
3 A Spontaneous Null Mutation of Fig4: The Pale Tremor Mouse
4 Tissue-Specific Fig4 Transgenes: Neurons Versus Astrocytes
5 Conditional Knockout of Fig4 in Neurons
6 The Human Disease Mutation FIG4-I41T in Transgenic Mice
7 A Spontaneous Missense Mutation of Vac14 in the ingls Mouse
8 A Null Gene-Trap Allele of Vac14
9 A Hypomorphic Gene-Trap Allele of Pikfyve (Fab1)
10 A Conditional Knockout of Pikfyve
11 Genetic Interactions: Fig4, Vac14, and Mtmr2
12 Genetic Effects of Strain Background
13 Future Applications of Mouse Models of PI(3,5)P2 Deficiency
Acknowledgments
References
Chapter Fifteen. Monitoring Endosomal Trafficking of the G Protein-Coupled Receptor Somatostatin Receptor 3
Abstract
1 Introduction
2 Development of Cell Lines Stably Expressing SSTR3
3 Live Imaging of SSTR3 and RABS in Mammalian Kidney Cells
4 Dynamics of SSTR3 Transit Relative to RABS
5 Effects of Dominant Negative Rabs on SSTR3 Trafficking
6 Summary
Acknowledgments
References
Section III: Proteins
Chapter Sixteen. Genetic Circuitry Modulating Notch Signals Through Endosomal Trafficking
Abstract
1 Introduction
2 Genetic Screen Using the Exelixis Collection
3 Notch Localization in Endosomes
4 Optical Approaches
5 Ubiquitination Status of Notch
6 Conclusion
Acknowledgments
References
Chapter Seventeen. Monitoring Notch/Delta Endosomal Trafficking and Signaling in Drosophila
Abstract
1 Introduction
2 Antibody Uptake Assays to Monitor Notch and Delta Trafficking
3 Correlative Imaging of Dividing SOP Cells and of Their Progeny
4 Concluding Remarks
Acknowledgements
References
Chapter Eighteen. Toll-Interacting Protein Pathway: Degradation of an Ubiquitin-Binding Protein
Abstract
1 Introduction
2 Expression of GFP-htt Protein and RFP-Tollip and How to Count Aggregation
3 Cytoskeleton-Dependent Transport of Tollip to httpQ-Aggresome
4 Determinants of Early-/Late Endosomal Localization in the Cells
5 Summary
Acknowledgment
References
Chapter Nineteen. Measuring Interactions of FERM Domain-Containing Sorting Nexin Proteins with Endosomal Lipids and Cargo Molecules
Abstract
1 Introduction
2 Secondary Structure-Based Domain Classification
3 Production of Recombinant PX-FERM Proteins
4 Measuring Interactions of PX-FERM Proteins with Phosphoinositide Lipids and Peptide Cargo Motifs by Isothermal Titration Calorimetry
5 Analysis of SNX17 PX Domain-PI3P Interactions by Nuclear Magnetic Resonance Spectroscopy
6 Summary
Acknowledgments
References
Author Index
Subject Index
Product details
- No. of pages: 412
- Language: English
- Copyright: © Academic Press 2014
- Published: December 17, 2013
- Imprint: Academic Press
- Hardcover ISBN: 9780123979261
- eBook ISBN: 9780123984821