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In the past several years significant attention has been given to the analysis of the properties and functions of lateral microdomains (rafts) in biological membranes. As described in the overview chapter of this book, as well as in the introductions to many of the other chapters, there are many fundamental un- swered questions concerning the composition, structure, dynamics, and even the very existence of these membrane rafts. Therefore, a variety of sophisticated techniques have been used to study intact cell membranes, as well as model s- tems composed of specific lipids and proteins thought to be in rafts. The analyses of biological membranes have provided detailed data on intact microdomains, and the complementary studies of model systems have given critical insights on the roles of specific lipid-lipid and lipid-protein interactions in raft formation and function. The chapters in this book provide detailed information on modern methods that are currently being employed to study lipid rafts.
List of contents
Overview of Membrane Rafts.- Analysis of Raft Affinity of Membrane Proteins by Detergent-Insolubility.- Nondetergent Isolation of Rafts.- Detecting Ordered Domain Formation (Lipid Rafts) in Model Membranes Using Tempo.- Using Monomolecular Films to Characterize Lipid Lateral Interactions.- Electro-Formation and Fluorescence Microscopy of Giant Vesicles With Coexisting Liquid Phases.- Fluorescence Correlation Spectroscopy.- Multiphoton Laser-Scanning Microscopy and Spatial Analysis of Dehydroergosterol Distributions on Plasma Membrane of Living Cells.- NMR Detection of Lipid Domains.- Lateral Diffusion Coefficients of Raft Lipids From Pulsed Field Gradient NMR.- Saturation-Recovery Electron Paramagnetic Resonance Discrimination by Oxygen Transport (DOT) Method for Characterizing Membrane Domains.- Plasmon-Waveguide Resonance Spectroscopy Studies of Lateral Segregation in Solid-Supported Proteolipid Bilayers.- Fluorescence Recovery After Photobleaching Studies of Lipid Rafts.- Single-Molecule Tracking.- X-Ray Diffraction to Determine the Thickness of Raft and Nonraft Bilayers.- Small-Angle Neutron Scattering to Detect Rafts and Lipid Domains.- Exploring Membrane Domains Using Native Membrane Sheets and Transmission Electron Microscopy.- Atomic Force Microscopy of Lipid Domains in Supported Model Membranes.- Atomistic and Coarse-Grained Computer Simulations of Raft-Like Lipid Mixtures.- A Microscopic Model Calculation of the Phase Diagram of Ternary Mixtures of Cholesterol and Saturated and Unsaturated Phospholipids.
Summary
In the past several years significant attention has been given to the analysis of the properties and functions of lateral microdomains (rafts) in biological membranes. As described in the overview chapter of this book, as well as in the introductions to many of the other chapters, there are many fundamental un- swered questions concerning the composition, structure, dynamics, and even the very existence of these membrane rafts. Therefore, a variety of sophisticated techniques have been used to study intact cell membranes, as well as model s- tems composed of specific lipids and proteins thought to be in rafts. The analyses of biological membranes have provided detailed data on intact microdomains, and the complementary studies of model systems have given critical insights on the roles of specific lipid-lipid and lipid-protein interactions in raft formation and function. The chapters in this book provide detailed information on modern methods that are currently being employed to study lipid rafts.
