General Principles and Procedures

General Principles and Procedures

by R.D. O'Brien (Editor)

Paperback(Softcover reprint of the original 1st ed. 1979)

View All Available Formats & Editions
Choose Expedited Shipping at checkout for guaranteed delivery by Thursday, May 2

Product Details

ISBN-13: 9781468409819
Publisher: Springer US
Publication date: 12/24/2012
Series: The Receptors , #1
Edition description: Softcover reprint of the original 1st ed. 1979
Pages: 345
Product dimensions: 7.01(w) x 10.00(h) x 0.03(d)

Table of Contents

1 Reconstitution of Membrane Transport Functions.- 1. Introduction.- 2. Reconstitution of Active and Passive Transport Systems.- 3. General Techniques of Reconstitution.- 3.1. Liposomes: Test Tubes with a Difference.- 3.1.1. Multilamellar Liposomes.- 3.1.2. Unilamellar Liposomes.- 3.2. Methods for Inserting Proteins into Liposomes.- 3.2.1. Cholate Dialysis.- 3.2.2. Sonication.- 3.2.3. Incorporation.- 3.2.4. The Use of Superstable Membrane Proteins.- 4. What We Can Learn from Reconstitution.- 4.1. Oxidative Phosphorylation.- 4.2. Ca2+-ATPase.- 4.3. (Na+ + K+)-ATPase.- 4.4. Acetylcholine Receptor.- 4.5. The Problem of Orientation.- 5. Reconstitution in Planar Bilayer Membranes.- 5.1. Sucrase-Isomaltase Complex.- 5.2. Acetylcholine Receptor in Planar Bilayers.- 5.3. Insertion of Whole Membrane Vesicles.- 5.4. Proton Pumps.- References.- 2 The Pharmacon-Receptor-Effector Concept: A Basis for Understanding the Transmission of Information in Biological Systems.- 1. Introduction.- 2. Biological Action.- 3. Receptors and Receptor Sites.- 4. Pharmacon-Receptor Interaction.- 5. Spare Receptors.- 6. Structure and Action.- 7. Accessory Receptor Sites.- 8. Steric Structure and Action.- 9. Selectivity in Action.- 10. Differentiation in Closely Related Receptor Types.- 11. Receptor Binding and Receptor Isolation.- 12. Dualism in Receptors for Agonists and Their Competitive Antagonists.- 13. The Aggregation-Segregation Concept.- 14. Dual Receptor Model.- 15. Combination of Pharmaca.- 16. The Slope of the Concentration-Effect Curves.- 17. The Allosteric Receptor Model.- 18. Binding and Displacement on Two or More Independent Classes of Receptor Sites.- 19. Two-Site Model.- 20. Reflection.- References.- 3 The Link between Drug Binding and Response: Theories and Observations.- 1. The Response to Acetylcholine-Like Drugs.- 1.1. Methods of Investigation of the Response.- 1.2. The Nature of the Response to Acetylcholine.- 1.3. The Response-Concentration Curve at Equilibrium.- 1.4. The Kinetics of the Response.- 1.4.1. Relationship between Methods of Studying Kinetics.- 1.4.2. Concentration-Jump Studies.- 1.4.3. Fluctuation Analysis.- 1.4.4. Voltage-Jump Relaxation Studies.- 1.5. Anesthetics, Local Anesthetics, and Channel Blocking.- 2. The Binding of Drugs to Acetylcholine Receptors.- 2.1. Methods for Investigation of Binding.- 2.2. Binding at Equilibrium.- 2.2.1. Cooperativity in Binding.- 2.2.2. Is There a Single Sort of Binding Site?.- 2.2.3. Binding to Junctional and Extrajunctional Receptors in Muscle.- 2.3. The Kinetics of Acetylcholine Binding.- 3. The Link between Drug Binding and Response.- 3.1. What Should a Mechanism Explain?.- 3.2. Some Mechanisms.- 3.3. The Concentration Dependence of Binding and Response at Equilibrium.- 3.4. The Nature of Efficacy, Partial Agonists, and Desensitization….- 3.5. Kinetics and Mechanism.- 3.5.1. What Does the Observation of a Single Time Constant Imply?.- 3.5.2. What Is the Rate-Limiting Step?.- 3.5.3. Concentration Dependence of Time Constants from Kinetic Studies.- 3.6. What Is the Origin of Voltage Dependence?.- 3.7. High Affinity Versus High Speed.- References.- 4 Kinetics of Cooperative Binding.- 1. Overview.- 2. General Introduction.- 3. Model I: koff as a Linear Function of Occupancy.- 3.1. Assumptions.- 3.2. Properties of the Model.- 3.2.1. Equilibrium.- 3.2.2. Association Curves.- 3.2.3. Dissociation Curves.- 3.3. Discussion.- 4. Application to the Insulin-Receptor System.- 4.1. The Controversy.- 4.2. Experimental Design.- 4.3. Simulation Results.- 4.4. Discussion.- 5. Model II: kon as a Linear Function of Occupancy.- 5.1. Introduction.- 5.2. The Model.- 5.3. Properties of the Model.- 5.3.1. Equilibrium.- 5.3.2. Association Curves.- 5.3.3. Dissociation Curves.- 5.4. Testing for Positive Cooperativity.- 6. General Discussion.- 7. A Guide to the Experimentalist.- Appendix A: Model I: Differential Equations and Solutions.- Appendix B: Addition of Fresh (Empty) Receptors.- Appendix C: Model II: Labeled Ligand Only.- Appendix D: Model II: Labeled and Unlabeled Ligands.- Appendix E: Optimization of Testing for Model II with ? > 0.- References.- 5 Distinction of Receptor from Nonreceptor Interactions in Binding Studies.- 1. Defining a Pharmacologic Receptor.- 2. Criteria for Receptor Interactions.- 3. The Problem of Relating Binding to Biological Responsiveness.- 4. Nonspecific Binding: Definition and Examples of Complications of Binding Data Analysis.- 5. Estimating the Affinity of the Unlabeled Ligand.- 6. Examples of Receptor-Like Nonreceptor Interactions.- 7. Conclusion.- References.- 6 Incorporation of Transport Molecules into Black Lipid Membranes.- 1. Introduction.- 2. Methodology.- 2.1. Formation and Composition of BLMs.- 2.2. Electrical Properties of BLMs.- 3. Mechanisms of Ion Permeability.- 3.1. Carriers.- 3.2. Channel Formers.- 4. Models of Interactions of Proteins with BLMs.- 5. Ionophorous Properties in BLMs of Functional Transport Molecules.- 5.1. Ca2+-ATPase: Dissection of a Transport System.- 5.2. (Na+ + K+)-ATPase.- 5.3. The Acetylcholine Receptor.- 6. The BLM as a Test System for Ionophorous Function of Isolated Membrane Proteins.- 6.1. Mitochondrial Membrane Proteins.- 6.2. Red Blood Cell Membrane Proteins.- 6.3. Gastric Mucosal Membrane Proteins.- 6.4. Dopamine-?-Hydroxylase.- 6.5. Rhodopsin.- 6.6. Immune Cytotoxic Factors.- 7. Coda.- References.- 7 Visualization and Counting of Receptors at the Light and Electron Microscope Levels.- 1. Receptors at the Cell Membrane.- 1.1. Introduction.- 1.2. Information Required on the Distribution of Receptors.- 2. The Labeling of Receptors for Localization.- 2.1. Approaches.- 2.2. Methods of Labeling and Visualizing Receptors.- 2.2.1. Autoradiography.- 2.2.2. Electron-Dense Label Attachment.- 2.2.3. Enzymatic Reaction Product Markers.- 2.2.4. Fluorescent Markers.- 2.2.5. X-Ray Microanalysis.- 2.3. Ligands for Receptor Labeling.- 2.3.1. Selection of Primary Ligands.- 2.3.2. Ligands Available for Receptor Tracing.- 3. Cell and Tissue Autoradiography.- 3.1. Problems of Application of a Labeled Ligand.- 3.1.1. Mode of Application.- 3.1.2. Nonspecific Labeling.- 3.1.3. Tissue Processing for Autoradiography of Receptors.- 3.1.4. Application to a Pseudo-Irreversible Reaction at a Receptor.- 3.2. Autoradiographic Methods.- 3.2.1. Treatments in Aqueous Media.- 3.2.2. Dry-Mount Methods.- 3.3. Interpretation of EM Autoradiographic Data on Receptors.- 3.3.1. Assignment of Silver Grains in Autoradiographs to Most Probable Locations of the Labeled Receptors.- 3.3.2. Calculation of Receptor Density.- 3.3.3. Isotopes and Resolution.- 3.4. Applications to Synaptic Receptors.- 4. Counting Receptors per Cell or per Synapse.- 4.1. Light Microscope Autoradiography of Receptors.- 4.2. Absolute Enumeration of Total Receptors.- 4.3. Direct Determination of Receptor Occupancy Relations.- 5. Electron Microscope Methods for Visualization of Receptors.- 5.1. Peroxidase Cytochemistry of Receptors.- 5.1.1. Peroxidase Methods.- 5.1.2. Applications of Peroxidase Cytochemistry to Receptors.- 5.2. Tissue Preservation for Immunocytochemistry of Receptors.- 5.3. Ferritin Labeling.- 5.4. Other Labels Applicable for Transmission and Scanning EM Studies of Receptors.- 6. Fluorescence Marker Methods.- 6.1. Fluorescence Labeling.- 6.2. Application of Fluorescent Labeling to Receptors.- 7. Possibilities of Quantitation of Receptors in Immunocytochemical and Other Nonradioisotopic Techniques.- 7.1. Quantitation in Electron-Dense Marker Techniques.- 8. Conclusions.- References.- 8 Problems and Approaches in Noncatalytic Biochemistry.- 1. Introduction.- 2. Measurement.- 2.1. Histological Techniques.- 2.2. Physical Separation: General Considerations.- 2.3. Equilibrium Dialysis.- 2.4. Other Physical Separations.- 2.5. Negative Binding.- 2.6. The Magnitude of the Off-Time.- 3. Relation of in Vivo to in Vitro Properties.- 3.1. Reversibility.- 3.2. Location.- 3.3. Specificity.- 3.4. Dissociation Constants.- 3.5. Detergents.- References.

Customer Reviews

Most Helpful Customer Reviews

See All Customer Reviews