Lipids

Lipids

1st Edition - February 16, 2012
  • Editors: Gilbert Di Paolo, Markus Wenk
  • eBook ISBN: 9780123864888
  • Hardcover ISBN: 9780123864871

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Description

Lipids are a broad group of naturally occurring molecules which includes fats, waxes, sterols, fat-soluble vitamins (such as vitamins A, D, E and K), monoglycerides, diglycerides, phospholipids, and others. The main biological functions of lipids include energy storage, as structural components of cell membranes, and as important signaling molecules.This volume of Methods in Cell Biology covers such areas as Membrane structure and dynamics, Imaging, and Lipid Protein Interactions. It will be an essential tool for researchers and students alike.

Key Features

  • Covers such areas as membrane structure and dynamics, imaging, and lipid protein interactions
  • An essential tool for researchers and students alike
  • International authors
  • Renowned editors

Readership

Researchers and students in cell, molecular and developmental biology

Table of Contents

  • Contributors

    Preface

    Part I: Membrane Dynamics and Reconstitution Assays

    Chapter 1: Supported Native Plasma Membranes as Platforms for the Reconstitution and Visualization of Endocytic Membrane Budding

    I. Introduction

    II. Preparation of Membrane Sheets

    III. Preparation of Brain Extract

    IV. Cell-Free Reaction

    V. Conclusions

    Chapter 2: Studying Lipids Involved in the Endosomal Pathway

    I. Introduction

    II. Methods

    III. Conclusion

    Appendix A. Supplementary Movies

    Chapter 3: Studying In Vitro Membrane Curvature Recognition by Proteins and its Role in Vesicular Trafficking

    I. Introduction

    II. Preparation of proteins and liposomes

    III. Binding Assays for Testing Curvature Recognition by a Protein

    IV. Distribution of a Curvature-Sensing Protein on Tube Networks Pulled by Kinesin Motors

    V. Distribution of a Curvature-Sensing Protein on a Tube Elongated by Optical Tweezers

    VI. Assays to Measure the Curvature-Dependant Activity of ArfGAPl and GMAP-210

    VII. Summary and Conclusion

    Chapter 4: Reconstituting Multivesicular Body Biogenesis with Purified Components

    I. Introduction

    II. Rationale

    III. Methods

    IV. Materials

    V. Discussion

    VI. Summary and Outlook

    Chapter 5: Approaches to the Study of Atg8-Mediated Membrane Dynamics In Vitro

    I. Introduction

    II. Using Liposomes as In Vitro Mimics of Autophagosome Membrane

    III. Proteins

    IV. The Lipidation Reaction

    V. Alternative Lipidation Approach for PE and Enzyme Independence

    VI. Membrane Tethering

    VII. Conclusion

    Chapter 6: Reconstitution Assay System for Ceramide Transport With Semi-Intact Cells

    I. Introduction

    II. Materials

    III. Methods

    IV. Notes

    Chapter 7: Visualizing Mitochondrial Lipids and Fusion Events in Mammalian Cells

    I. Introduction

    II. Imaging Mitochondrial Tubules in Overexpression or Knockdown Samples

    III. A Quantitative Assay for Mitochondrial Fusion

    IV. Visualizing Lipids in Cells

    V. Summary

    Part II: Lipid Metabolism and Signaling

    Chapter 8: Targeted and Non-Targeted Analysis of Membrane Lipids Using Mass Spectrometry

    I. Introduction

    II. Isolation and Purification of Membrane Lipids

    III. Detailed Protocols for Isolation of Membrane Lipid From Mammalian Cells and Tissues

    IV. Mass Spectrometry-Based Approaches for Lipid Analysis

    A. Details of Lipidomics Analysis

    Chapter 9: Modulation of Host Phosphoinositide Metabolism During Salmonella Invasion by the Type III Secreted Effector SopB

    I. Introduction

    II. Rationale

    III. Materials and Media

    IV. Methods

    V. Summary and Conclusions

    Chapter 10: Acute Manipulation of Phosphoinositide Levels in Cells

    I. Introduction

    II. Rationale

    III. Preparation of Expression Constructs

    IV. Expression of Fusion Proteins and Cell Maintenance

    V. Microscopy

    VI. Considerations

    VII. Summary and Conclusion

    Chapter 11: Regulation of Phosphoinositide-Metabolizing Enzymes by Clathrin Coat Proteins

    I. Introduction

    II. Inducible Expression of PI-Metabolizing Enzymes

    III. Radioactive Kinase Activity Assay

    IV. Interpretation and Troubleshooting

    V. Outlook

    Chapter 12: Phosphoinositides at the Neuromuscular Junction of Drosophila melanogaster: A Genetic Approach

    I. Introduction

    II. Genetic Tools to Study Phosphoinositides in Drosophila

    III. Cellular Processes Regulated by Phosphoinositides at the Fly NMJ

    IV. Future Directions

    V. Conclusions

    Chapter 13: Devising Powerful Genetics, Biochemical and Structural Tools in the Functional Analysis of Phosphatidylinositol Transfer Proteins (PITPs) Across Diverse Species

    I. Introduction

    II. Rationale

    III. In Vitro Approaches

    IV. In Vivo Approaches

    V. Structural Approach

    VI. Conclusions and Summary

    Chapter 14: Genome-Wide Screens for Gene Products Regulating Lipid Droplet Dynamics

    I. Introduction

    II. Genome-Wide Screen of Yeast Deletion Mutants for Changes in the Dynamics of Lipid Droplets

    III. Additional Insights From Genome-Wide Studies in Drosophila Cells

    IV. Concluding Remarks

    Chapter 15: The Three Dimensionality of Cell Membranes: Lamellar to Cubic Membrane Transition as Investigated by Electron Microscopy

    I. Introduction

    II. Cell Models to Study Non-lamellar Membrane Organizations

    III. Understanding Highly Ordered Membrane Arrangements through Transmission Electron Microscopy and Computer Simulation

    IV. A Closer Look at Cubic Membrane Surface Contours through Scanning Electron Microscopy (SEM)

    V. Summary

    Chapter 16: Quantitative Imaging of Lipid Metabolism in Yeast: From 4D Analysis to High Content Screens of Mutant Libraries

    I. Introduction

    II. Choice of Fluorescence Dyes for LD Labeling

    III. Four-Dimensional Live Cell Imaging of Yeast LD During Cellular Growth

    IV. Imaging-Based Quantitative Analysis of Yeast LD in Large Cell Populations

    V. Label Free Imaging of yeast LD using CARS Microscopy

    VI. Summary and Conclusions

    Chapter 17: Analysis of Cholesterol Trafficking with Fluorescent Probes

    I. Introduction

    II. Concluding remarks

    Chapter 18: Fluorescence Correlation Methods for Imaging Cellular Behavior of Sphingolipid-Interacting Probes

    I. Introduction

    II. Rationale

    III. Methods

    IV. Materials

    V. Discussion

    VI. Summary and Outlook

    Chapter 19: Monitoring Phospholipid Dynamics during Phagocytosis: Application of Genetically-Encoded Fluorescent Probes

    I. Introduction

    II. Rationale

    III. Materials

    IV. Methods

    V. Considerations when Designing an Experiment

    VI. Summary

    Chapter 20: Genetically Encoded Probes for Phosphatidic Acid

    I. Phosphatidic Acid: A Rapid Overview

    II. Choice of the PA-Probes

    III. Specific Binding of PA to Probes

    IV. Imaging PA in Cells

    V. Summary and Conclusion

    Index

    VOLUMES IN SERIES

Product details

  • No. of pages: 528
  • Language: English
  • Copyright: © Academic Press 2012
  • Published: February 16, 2012
  • Imprint: Academic Press
  • eBook ISBN: 9780123864888
  • Hardcover ISBN: 9780123864871

About the Serial Volume Editors

Gilbert Di Paolo

Markus Wenk