Methods in Membrane Biology: Volume 8

1 The Use of Organic Solvents in Membrane Research.- 1. Introduction.- 1.1. General Considerations.- 1.2. Solvolysis and Solvation.- 1.3. Effect of Organic Solvents on Water.- 1.4. Effect of Organic Solvents on Lipids.- 1.5. Effect of Organic Solvents on Proteins.- 2. Applications of Organic Solvents in Membranology.- 2.1. Chloroform-Methanol.- 2.2. Acetone.- 2.3. Ethyl Alcohol.- 2.4. Diethyl Ether.- 2.5. Butanol.- 2.6. Acetic and Formic Acid.- 2.7. 2-Chloroethanol.- 2.8. Phenol and Pyridine.- 2.9. Miscellaneous.- 3. Appendix.- Method 1: Isolation of Total Lipids from Animal Tissues by Chloroform-Methanol.- Method 2: Extraction and Purification of Lipids from Animal Tissues by Chloroform-Methanol.- Method 3: Extraction of Serum Phosphatides by Chloroform-Methanol.- Method 4: Extraction and Solubilization of Mammalian Erythrocyte Membrane Glycoproteins by ChloroformMethanol-Water.- Method 5: Extraction of Lipids from Blood with an Ethanol-Ether Mixture.- Method 6: Extraction of Loosely and Strongly Bound Lipids from Erythrocyte Membranes by Diethyl Ether and Ethanol.- Method 7: A Modified n-Butanol Extraction of Erythrocyte Membranes.- Method 8: Solubilization of Erythrocyte Membrane Proteins by Acetic Acid.- Method 9: Solubilization and Isolation of Erythrocyte Membrane Protein by 2-Chloroethanol.- Method 10: Recombination of Erythrocyte Membrane Proteins and Lipids in 2-Chloroethanol.- Method 11: Isolation of Glycoproteins from Cell Membranes with Lithium Diiodosalicylate (LIS).- Method 12: Solubilization of Erythrocyte Membrane Proteins with Aqueous Pyridine.- Method 13: Delipidation of a Proteolipid Protein from Cerebral Cortex Using Sephadex LH-20 and Elution with N,N-Dimethylformamide-HCl.- 4. References.- 2 Recent Methods for the Elucidation of Lipid Structure.- 1. Introduction.- 2. Separation Techniques.- 2.1. Extraction of Tissue.- 2.2. Fractionation by Lipid Class.- 2.3. Gas-Liquid Chromatography.- 2.4. Fractionation by Molecular Property.- 3. Chemical Identification.- 3.1. Degradation of Complex Lipids.- 3.2. Recovery and Separation of Cleavage Products.- 3.3. Analysis of Constituents.- 3.4. Reagents for Detecting Specific Functional Groups.- 3.5. Positional Analysis.- 3.6. Assessment of Purity.- 4. The Analysis of Stereoisomers.- 4.1. Geometrical Isomerism.- 4.2. The Determination of Optical Configuration and Enantiomeric Purity.- 5. Mass Spectrometry of Lipids.- 5.1. Introduction.- 5.2. The Ionization Process.- 5.3. The Quasi-Equilibrium Theory (QET) of Mass Spectra.- 5.4. Ionization and Appearance Potentials.- 5.5. Experimental Predictions of the QET.- 5.6. Ionization Techniques.- 5.7. The Mass Spectrum.- 5.8. Applications.- 5.9. Wax Esters.- 5.10. Glycerides and Diol Esters.- 5.11. Complex Lipids-Glycolipids, Sphingolipids, Phospholipids, and Phosphonolipids.- 5.12. Sterols.- 5.13. Prostaglandins.- 5.14. Miscellaneous.- 6. Proton Nuclear Magnetic Resonance Spectroscopy of Lipids.- 6.1. Introduction.- 6.2. Spin-Spin Coupling and Conformational Studies.- 6.3. Quantitative Analytical Applications of High Resolution NMR.- 6.4. Low Resolution (Wide Band) NMR.- 6.5. NMR Chemical Shift Reagents.- 7. Carbon-13 Nuclear Magnetic Resonance Spectroscopy.- 7.1. Introduction.- 7.2. The Prediction of 13C Chemical Shift Parameters.- 7.3. Applications.- 8. Artifacts and Contaminants.- 9. References.- 3 Synthesis of Stereoisomeric Phospholipids for Use in Membrane Studies.- 1. Introduction and Historical Survey.- 2. Procedures for Synthesis of Acyl Phospholipids.- 2.1. Starting Materials.- 2.2. Phosphatidylcholines (PC).- 2.3. Phosphatidylethanolamines (PE).- 2.4. N-Methyl-and N,N-Dimethyl-3-sn-phosphatidylethanolamines.- 2.5. 3-sn-Phosphatidylserine (PS).- 2.6. Phospholipids with Unusual Nitrogenous Bases.- 2.7. Phosphatidylglycerols (PG) and Derivatives.- 2.8. Bisphosphatidic Acids (Bis-PA), Semi-lysobisphosphatidic Acid (Semi-lyso-bis-PA) and Lyso-bisphosphatidic Acid (Lyso-bis-PA).- 2.9. Diphosphatidylglycerol (Cardiolipin