Methods in Membrane Biology

1 Isolation of Cell Surface Membrane Fractions from Mammalian Cells and Organs.- 1. Introduction.- 2. Fractionation Characteristics of Cellular Organelles.- 2.1. Background.- 2.2. Nuclei.- 2.3. Mitochondria.- 2.4. Endoplasmic Reticulum.- 2.5. Golgi Apparatus.- 2.6. Glycogen.- 2.7. Fat Droplets.- 2.8. Cell Surface Membrane.- 3. General Types of Procedures.- 4. Evaluating Surface Membrane Preparations.- 5. Relation between Isolated Surface Membrane and in Vivo Membrane.- 6. Explanation of Table I.- 7. Brief Summary of Table I.- 8. References.- 2 Methods for the Isolation and Structural Characterization of Hepatocyte Gap Junctions.- 1. Introduction.- 1.1. The Desmosome.- 1.2. The Zonula Occludens.- 1.3. The Gap Junction (Nexus).- 2. Methods.- 2.1. Isolation of Liver Gap Junctions.- 2.2. Electron Microscopy.- 2.3. Nomenclature.- 2.4. Splitting of Hepatocyte Gap Junctions in Whole Liver.- 2.5. X-Ray Diffraction.- 3. References.- 3 Membrane Receptors for Polypeptide Hormones.- 1. Introduction.- 2. Studies of Peptide Hormone Receptors.- 2.1. Indirect Studies.- 2.2. Early Direct Studies.- 3. Methodology.- 3.1. Labeled Hormones.- 3.2. Receptor Preparations.- 3.3. Methods of Separation.- 4. Characteristics of the Receptor.- 5. Quantitative Aspects of Hormone-Receptor Studies.- 5.1. Equilibrium Studies.- 5.2. Kinetic Analysis.- 5.3. Cooperativity in Binding.- 5.4. Correlation of Binding and Biological Effect.- 5.5. Hormone and Receptor Degradation.- 6. Chemical Characterization of Hormone Receptors.- 7. Applications of Hormone-Receptor Studies.- 7.1. Mechanism of Hormone Action and Structure-Activity Relationships for Peptide Hormones.- 7.2. Assay of Plasma Hormones.- 7.3. Role of the Receptor in States of Altered Hormone Sensitivity.- 7.4. Factors Regulating Hormone Receptors.- 8. Future Directions.- 9. References.- 4 Use of Lectins for the Study of Membranes.- 1. Introduction.- 2. General Properties of Lectins.- 3. Methods for Study of Binding of Lectins to Cell Surfaces.- 3.1. Radioactively Labeled Lectins.- 3.2. Microscopic Techniques.- 4. Distribution of Lectin Receptor Sites.- 5. Effects of Binding of Lectins to Cells.- 5.1. Agglutination.- 5.2. Mitogenic Stimulation.- 5.3. Other Effects.- 6. Cell Receptors for Lectins.- 6.1. Isolation of Receptors.- 6.2. Fractionation and Purification of Receptors.- 7. References.- 5 Turnover of Membrane Proteins in Animal Cells.- 1. Introduction.- 2. Methods for Studying Turnover.- 2.1. A Simple Model for the Description of Changes in Protein Levels.- 2.2. Methods Based on Isotope Incorporation and Decay.- 2.3. Use of Surface-Labeling Reagents for the Study of Turnover of Membrane Proteins.- 2.4. Methods Based on Kinetics of Change in Enzyme Activity.- 3. General Properties of Turnover of Proteins.- 4. On the Mechanisms of Protein Turnover in Animal Cells.- 4.1. Properties of the Protein Molecule as a Substrate for Degradation.- 4.2. Alterations in Activity of a Degradative Process.- 5. Turnover of Membrane Proteins.- 6. Mechanisms of Genesis and Turnover of Membranes in Animal Tissues.- 7. Conclusion.- 8. References.