Description

This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This is the first of two volumes on endosome signaling and includes chapters on such topics as measurement of entry into the endosomal compartment by multi-parametric image analysis, assessment of peptide internalization and endosomal signaling, and VEGF-A in endosomal signaling.

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 Ev

Details

No. of pages:
412
Language:
English
Copyright:
© 2014
Published:
Imprint:
Academic Press
Print ISBN:
9780123979261
Electronic ISBN:
9780123984821