Intracellular Lipid Transport Methods and Protocols

This fully updated volume explores different classes of lipid transporters, such as lipid transfer proteins, flippases, and scramblases, and significant advances in solving their biological functions.  The first half of the book features methodologies to measure the movement of lipids between or in organelle membranes inside eukaryotic cells, including plant cells, or in prokaryotic cells, and the book continues with in vitro or in silico approaches aiming to define, more from a biochemical and structural standpoint, how lipid transfer proteins function. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and up-to-date, Intracellular Lipid Transport: Methods and Protocols, Second Edition serves as an ideal guide for researchers seeking to shed light on diverse aspects of this critical and often elusive cellular process.

Selective Labeling of Phosphatidylcholine to Track Their Translocation Between Organelles in Living Cells.- Measurement of ORP10-Mediated Lipid Countertransport at ER-Endosome Membrane Contact Sites Via a Chemically-Induced Dimerization Strategy.- Analysis of Phosphatidylinositol Transfer at ER-PM Contact Sites in Receptor-Stimulated Live Cells.- Use of the D4H Probe to Track Sterols in Yeast.- Maistero-2, a Novel Probe for Sterols: Application for Visualizing Cellular Cholesterol.- Using GRAM Domain-Based Biosensors to Monitor Accessible Cholesterol Distribution in Live Cells.- Monitoring Accessible Cholesterol Levels in Immune Cells.- Methods for Visualizing and Quantifying Cholesterol Distribution in Mammalian Cells Using Filipin and D4 Probes.- HEK293 Cell-Based Assay to Measure the Lipid Scrambling Activity of TMEM16 Family Members.- Measuring Phospholipid Flippase Activity by NBD-Lipid Uptake in Living Yeast Cells.- Advanced Method for the In Vivo Measurements of Lysophospholipid Translocation Across the Inner (Cytoplasmic) Membrane of Escherichia coli.- Measurement of Lipid Transport in Mitochondria by the MTL Complex.- Determining ATG2 Localization During Autophagosome Formation in Mammalian Cells.- Use of Bicelle-Generated Lipid Bilayer Vesicle Nanoparticles for In Vitro Measurement of Lipid Intermembrane Transport.- DNA-Assisted Assays for Studying Lipid Transfer Between Membranes.- Galactocerebroside Lipid Nanotubes, a Model Membrane System for Studying Membrane-Associated Proteins on a Molecular Scale.- In Vitro Approaches for Detection of Protein-Lipid Interactions and Protein-to-Protein Transport of Lipids Using Bifunctional Lipid Analogs.- Determining the Relative Affinity of ORPs for Lipid Ligands Using Fluorescence and Thermal Shift Assays.- Atomistic Simulations and Analysis of Peripheral Membrane Proteins with Model Lipid Bilayers.- In Silico Strategies for Characterizing Inner Cavities of Lipid-Binding Proteins.