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 second 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: SIGNALING

Chapter One. Assessment of Insulin Proteolysis in Rat Liver Endosomes: Its Relationship to Intracellular Insulin Signaling

Abstract

1 Introduction

2 Assays for Insulin Degradation

3 Assay for Endosomal Proteolysis of Insulin

4 Sites of Cleavage and Degradation Products of Insulin within Endosomes

5 Endosomal Insulinases

6 Effects of Insulin and Slowly Processed Insulin Analogs on Insulin Signaling in Liver Endosomes

7 Involvement of IR Endocytosis in Positive Regulation of Insulin Signaling

References

Chapter Two. A Bimolecular Fluorescent Complementation Screen Reveals Complex Roles of Endosomes in Ras-Mediated Signaling

Abstract

1 Introduction

2 Detect Ras–effector Interactions by BiFC

3 Screen for Ras-binding Proteins by BiFC

4 Concluding Remarks

Acknowledgments

References

Chapter Three. TGFβ in Endosomal Signaling

Abstract

1 Introduction

2 Endocytosis

3 The Role of the Early Endosome in TGFβ Signal Transduction

4 Readouts of TGFβ Signal Transduction

Acknowledgments

References

Chapter Four. Annexins and Endosomal Signaling

Abstract

1 Introduction

2 Isolation of Endocytic Compartments

3 Targeting Raf-1 Signaling to Early Endosomes

4 Monitoring Endosomal Signaling by Fluorescence Resonance Energy Transfer Microscopy

5 Targeting Annexins to Endosomes and Other Cellular Compartments

6 Summary

Acknowledgments

References

Chapter Five. Analysis, Regulation, and Roles of Endosomal Phosphoinositides

Abstract

1 Introduction

2 Endosomal PIs

3 Analysis of PtdIns3P Levels

4 Monitoring PtdIns3P Intracellular Localization

5 Endosomal PtdIns3P

6 Phospha

Details

No. of pages:
518
Language:
English
Copyright:
© 2014
Published:
Imprint:
Academic Press
Print ISBN:
9780123979254
Electronic ISBN:
9780123984814