Membrane Analysis (Paperback)

Abbreviations -- Preface -- 1. General membrane composition and structure -- Lipids -- General lipid composition -- Phospholipids -- Isoprenoids -- Lipid architecture -- Proteins -- Peripheral proteins -- Integral proteins -- Lipid-anchored proteins -- Protein architecture in the plane of the membrane -- Carbohydrates -- Glycolipids -- Glycoproteins -- Proteoglycans -- 2. The membranes of prokaryotes and eukaryotes -- General features -- Membranes of prokaryotes -- Membranes of eukaryotes -- The surface (plasma) membrane -- Plasma membrane domains -- The cytoskeleton and glycocalyx -- Mitochondria and chloroplasts -- Peroxisomes (microbodies) -- Lysosomes -- Nuclei -- Endocytic compartment -- The secretory compartment -- References -- 3. Preparation of subcellular membranes -- Homogenization -- Liquid shear -- Gaseous shear -- Mechanical shear -- Other methods -- Homogenization of soft mammalian tissue -- Homogenization of tough tissues -- Homogenization of cultured mammalian cells -- Plant tissue -- Yeast -- Bacteria -- Fractionation of subcellular organelles -- Differential centrifugation -- Density gradient centrifugation -- Density perturbation -- Fractionation by immunoaffinity -- Fractionation by continuous flow electrophoresis -- Fractionation in aqueous two-phase systems -- Identification of fractionated membranes -- Morphological -- Chemical -- Enzyme markers -- Immunological markers -- Expression of data -- Some specimen separations of organelles and membranes -- Nuclei -- Golgi membranes -- Mitochondria -- Resolving a light mitochondrial fraction -- Plasma membrane -- Membrane vesicles -- References -- 4. Molecular composition -- Chemical analysis of proteins -- Quantitative assay -- Separation of membrane proteins by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) -- Western blotting (immunoblotting) -- Resolution of proteins by isoelectric focusing and twodimensional (2D) electrophoresis -- Chemical analysis of lipids -- Lipid extraction -- Analysis of phospholipids -- Separation and analysis of phospholipids by HPLC -- Analysis of phospholipid fatty acid -- Determination of cholesterol -- Separation and analysis of glycolipids -- Glycoprotein oligosaccharide analysis -- Glycoprotein analysis -- Preparation of a crude glycoprotein fraction -- Glycoprotein fractionation -- Chemical assays for carbohydrates -- Isolation of the oligosaccharide chains -- Oligosaccharide sequence analysis -- References -- 5. Investigating the topology of membrane proteins -- Demonstration of the orientation of membrane vesicles -- Identification and separation of peripheral and integral proteins -- Triton X114 as a reagent for separation of integral and peripheral proteins -- Identification of proteins anchored by phosphatidylinositol -- Identification of lipid-modified proteins -- Probing the topography of integral membrane proteins -- General experimental criteria -- Probes for integral membrane proteins -- Hydropathy plots for prediction of membrane protein topography -- Structural analysis of membrane proteins -- References -- 6. Investigation of membrane lipids -- Determination of the organization of membrane phospholipids -- Model membranes -- Multilamellar vesicles (MLVs) -- Small unilamellar vesicles (SUVs) -- Large unilamellar vesicles (LUVs) -- Black lipid membranes -- Reconstituted membranes -- Determination of the transverse distribution of membrane phospholipids -- Chemical probes -- Phospholipases -- Phospholipid transfer proteins -- Assay based on prothrombinase -- Determination of the transverse distribution of membrane cholesterol -- Studies of lipid transport -- Investigation of the transmembrane movement of phospholipids -- Transport of phospholipids and cholesterol between intracellular membranes -- Genetic approach to investigate membrane phospholipids -- References -- 7. Protein targetting and membrane biogenesis -- Genetic approaches to studies of protein targetting -- Biochemical approaches to studies of protein targetting -- Steps in protein targetting -- Nuclear targetting -- Targetting to peroxisomes -- Targetting to mitochondria -- Targetting to the ER -- Transport across the ER membrane -- Retention in the ER lumen -- Transfer to the Golgi apparatus -- Targetting to lysosomes -- Transit of mannose-6-phosphatase receptors to and from the plasma membrane and Golgi membranes -- Establishing the topography and location of the membrane proteins of the ER, Golgi and plasma membrane -- Covalently bound lipids in protein targetting -- Targetting in prokaryotes -- Overview -- References -- 8. Endocytosis and vesicular trafficking -- Receptors and endocytosis -- Receptor-mediated endocytosis -- Membrane receptors -- Investigation of events in endocytosis -- Electron microscopy -- Biochemical studies -- Isolation of transport vesicles and investigation of the factors involved in vesicular transport -- Reconstitution of vesicular transport -- Semi-intact cell systems -- Transfer vesicles -- Transfer from the plasma membrane to the endosomes -- Transfer from the Golgi to lysosomes -- Golgi to endoplasmic reticulum (retrograde transport) -- Endoplasmic reticulum to Golgi (anterograde transport) -- Recognition in vesicular transfer -- References -- Index.