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
eBook ISBN:
9780123984821
Print ISBN:
9780123979261

About the serial-volume-editor

P. Michael Conn

P. Michael Conn is the Senior Vice President for Research and Associate Provost, Texas Tech Health Sciences Center. He is The Robert C. Kimbrough, Professor of Internal Medicine and Cell Biology/Biochemistry. He was previously Director of Research Advocacy and Professor of Physiology and Pharmacology, Cell Biology and Development and Obstetrics and Gynecology at Oregon Health and Science University and Senior Scientist of the Oregon National Primate Research Center (ONPRC). He served for twelve years as Special Assistant to the President and Associate Director of the ONPRC. After receiving a B.S. degree and teaching certification from the University of Michigan (1971), a M.S. from North Carolina State University (1973), and a Ph.D. degree from Baylor College of Medicine (1976), Conn did a fellowship at the NIH, then joined the faculty in the Department of Pharmacology, Duke University Medical Center where he was promoted to Associate Professor in 1982. In 1984, he became Professor and Head of Pharmacology at the University of Iowa College of Medicine, a position he held for eleven years. Conn is known for his research in the area of the cellular and molecular basis of action of gonadotropin releasing hormone action in the pituitary and therapeutic approaches that restore misfolded proteins to function. His work has led to drugs that have benefitted humans and animals. Most recently, he has identified a new class of drugs, pharmacoperones, which act by regulating the intracellular trafficking of receptors, enzymes and ion channels. He has authored or co-authored over 350 publications in this area and written or edited over 200 books, including texts in neurosciences, molecular biology and endocrinology. Conn has served as the editor of many professional journals and book series (Endocrinology, Journal of Clinical Endocrinology and Metabolism, Endocrine, Methods, Progress in Molecular Biology and Translational Science and Contemporary Endocrinology). Conn serve

Affiliations and Expertise

Texas Tech University Health Sciences Center, Lubbock, USA