Methods in Membrane Biology: Volume 3 Plasma Membranes

ISBN-13: 9781475702675 / Angielski / Miękka / 2012 / 246 str.

Edward D. Korn

Opis

Volume 3 continues the approach carried out in the first two volumes of this se ries of publishing articles on membrane methodology which include, in addition to procedural details, incisive discussions of the ap- plications of the methods and of their limitations. Wh at is the theoretical basis of the method, how and to what problems can it be applied, how does one interpret the results, what has thus far been achieved by the method, what lies in the future-these are the questions the authors have tried to answer. No area of membrane biology engages the interest of more investigators than studies of the plasma membrane. Four chapters in this volume are concerned with one or more aspects of the cell surface. Fundamental to all studies of the cell surface are the isolation and characterization of pure plasma membranes. Many preparations described in the literature are inadequate or are inadequately characterized. In the first chapter, Neville discusses the theoretical and practical bases of tissue fractionation, empha- sizes the variations in enzyme content among plasma membranes from different sources, offers guidance in the choice of the proper criteria for assessing membrane purity, and suggests the best markers for detecting the possible presence of contaminating organelles. To review in detail each of the many preparations of plasma membranes that have been published is impossible.

Spis treści

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.