Endosome Signaling Part A

Endosome Signaling Part A

1st Edition - December 17, 2013

Write a review

  • Editor: P. Michael Conn
  • Hardcover ISBN: 9780123979261
  • eBook ISBN: 9780123984821

Purchase options

Purchase options
Available
DRM-free (PDF, Mobi, EPub)
Sales tax will be calculated at check-out

Institutional Subscription

Free Global Shipping
No minimum order

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 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

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 served on the National Board of Medical Examiners, including two years as chairman of the reproduction and endocrinology committee. The work of his laboratory has been recognized with a MERIT award from the NIH, the J.J. Abel Award of the American Society for Pharmacology and Experimental Therapeutics, the Weitzman, Oppenheimer and Ingbar Awards of the Endocrine Society, the National Science Medal of Mexico (the Miguel Aleman Prize) and the Stevenson Award of Canada. He is the recipient of the Oregon State Award for Discovery, the Media Award of the American College of Neuropsychopharmacology and was named a distinguished Alumnus of Baylor College of Medicine in 2012. Conn is a previous member of Council for the American Society for Cell Biology and the Endocrine Society and is a prior President of the Endocrine Society, during which time he founded the Hormone Foundation and worked with political leadership to heighten the public’s awareness of diabetes. Conn’s students and fellows have gone on to become leaders in industry and academia. He is an elected member of the Mexican Institute of Medicine and a fellow of the American Association for the Advancement of Science. He is the co-author of The Animal Research War (2008) and many articles for the public and academic community on the value of animal research and the dangers posed by animal extremism. His op/eds have appeared in The Washington Post, The LA Times, The Wall Street Journal, the Des Moines Register, and elsewhere. Conn consults with organizations that are influenced by animal extremism and with universities and companies facing challenges from these groups.

Affiliations and Expertise

Texas Tech University Health Sciences Center, Lubbock, USA

Ratings and Reviews

Write a review

There are currently no reviews for "Endosome Signaling Part A"