Fluorescence Studies on Biological Membranes

1 Biomembrane Structure and Dynamics Viewed by Fluorescence.- 1. Introduction to Fluorescence.- 2. Dynamics and Structure of Membranes.- 2.1. Lateral and Rotational Diffusion.- 2.2. Orientational Order and Packing.- 2.3. Asymmetry.- 2.4. Lipid Domains.- 3. Fluorescence Techniques and What They Make Visible.- 3.1. Fluorescence Depolarization.- 3.2. Quenching.- 3.3. Fluorescence Energy Transfer.- 3.4. Fluorescence Recovery after Photobleaching (FRAP).- 3.5. Excimer Fluorescence.- 4. Summary and Conclusions.- 5. References.- 2 Dynamic Structure of Membranes and Subcellular Components Revealed by Optical Anisotropy Decay Methods.- 1. Introduction.- 2. Optical Anisotropy Decay.- 2.1. Principle of Optical Anisotropy Decay Method.- 2.2. Experimental Techniques.- 2.3. Information Contained in an Anisotropy Decay.- 2.4. Optical Anisotropy Decay as a Tool in Bioscience.- 3. Examples of Application.- 3.1. Dynamic Structure of Membranes Probed by Diphenylhexatriene.- 3.2. Protein Rotations in Membrane and on Membrane Surface.- 3.3. Internal Motion of DNA.- 3.4. Internal Motion of Actin Filament.- 3.5. Dynamic Structure of Myosin Filament.- 4. Concluding Remarks.- 5. References.- 3 Principles of Frequency-Domain Fluorescence Spectroscopy and Applications to Cell Membranes.- 1. Introduction.- 2. Comparison of Time- and Frequency-Domain Measurements.- 2.1. A First-Generation Frequency-Domain Fluorometer.- 2.2. Resolution of a Two-Component Mixture.- 3. Theory of Frequency-Domain Fluorometry.- 3.1. Decays of Fluorescence Intensity.- 3.2. Decays of Fluorescence Anisotropy.- 4. Intensity Decays of DPH-Labeled Membranes.- 5. Anisotropy Decays of Labeled Membranes.- 5.1. Hindered Rotations of Diphenylhexatriene.- 5.2. Anisotropic Rotations of Perylene.- 6. Time-Resolved Emission Spectra.- 6.1. Calculation of Time-Resolved Emission Spectra.- 6.2. Time-Resolved Emission Centers of Gravity and Spectral Half-Widths.- 6.3. Time-Resolved Spectral Data for Patman-Labeled Membranes.- 7. Energy Transfer in Membranes.- 7.1. Distribution of Distances in a Covalently Linked Donor-Acceptor Pair.- 8. A 2-GHz Frequency-Domain Fluorometer.- 8.1. Picosecond Resolution of Tyrosine Intensity and Anisotropy Decays.- 8.2. Measurement of a 8-psec Correlation Time.- 9. Future Developments.- 10. Summary.- 11. References.- 4 Time-Resolved Fluorescence Depolarization Techniques in Model Membrane Systems: Effect of Sterols and Unsaturations.- 1. Introduction.- 2. Intrinsic Motional Properties of Some Widely Used Fluorescent Probes.- 2.1. Motional Characteristics.- 2.2. Excited-State Characteristics.- 3. Sterol-Phospholipid Interactions in Model Membranes.- 3.1. Cholesterol-Phospholipid Interactions: Lecithin as Bilayer Matrix.- 3.2. Cholesterol-Phospholipid Interactions: Phospholipids Other Than Lecithin as Bilayer Matrix.- 3.3. Cholesterol Chemical Modification: Effect on Phospholipid Fatty Acyl Chains Order and Dynamics.- 4. Concluding Remarks.- 5. References.- 5 Fluorescence Polarization to Evaluate the Fluidity of Natural and Reconstituted Membranes.- 1. Introduction.- 1.1. Aims and Scope of This Chapter.- 1.2. Mechanism of Action and Biological Significance of Fluorescence Polarization Measurements of Membrane Fluidity.- 2. Methodology.- 2.1. Theory of Fluorescence Polarization for Ion-Membrane Measurements.- 2.2. Probe-Membrane Interactions.- 2.3. Probe-Ion Interactions.- 3. Current Advancements in the Measurement of Ion-Membrane Interactions Using Fluorescence Polarization.- 3.1. Natural Membranes.- 3.2. Reconstituted Membranes.- 4. Critical Evaluation of the Significance of Ion-Membrane Measurements.- 4.1. Advantages of Fluorescence Polarization for Evaluation of Ion-Membrane Interactions.- 4.2. Limitations of Fluorescence Polarization for Measurement of Ion-Membrane Interactions.- 4.3. Substantiation of the Fluorescence Polarization Measurements of Ion-Membrane Interactions.- 5. Concluding Remarks.- 6. References.- 6 Fluidity of Thyroid Plasma Membranes.- 1. Introduction.- 2. Thyroid Plasma Membranes.- 2.1. Enriched Plasma Membrane Fractions.- 2.2. Chemical Characterization of Purified Plasma Membranes.- 2.3. Enzymic Characterization of Purified Plasma Membranes.- 2.4. Subfractionation of Thyroidal Plasma Membranes.- 2.5. Characterization of Thyroid Plasma Membrane Subfractions.- 3. Fluidity of Thyroid Plasma Membranes.- 3.1. Fluidity Measurements.- 3.2. Fluidity of Thyroid Subcellular Fractions.- 3.3. Fluidity of a P2 Fraction in Reconstituted Thyroid Plasma Membranes.- 3.4. Fluidity Characteristics of Plasma Membrane Subfractions.- 4. Modulation of the Adenylate Cyclase Activity by Manipulating the Plasma Membrane Composition.- 4.1. Incorporation of Phospholipids.- 4.2. Incorporation of Gangliosides.- 4.3. Incorporation of Dolichol and Dolichyl Derivatives.- 4.4. Addition of Membrane-Perturbing Drugs.- 5. Involvement of Membrane Fluidity on Human Normal and Pathological Thyroid Glands.- 6. References.- 7 Spectroscopic Analysis of the Structure of Bacteriorhodopsin.- 1. Introduction.- 2. Principle of the Fluorescence Energy Transfer Technique.- 3. Three-Dimensional Disposition of the Retinal Chromophore in the Purple Membrane.- 3.1. In-Plane Location.- 3.2. Transmembrane Location.- 3.3. Orientation of the Molecular Plane.- 4. In-Plane Location of NBD (7-Chloro-4-Nitrobenzo-2-Oxa-l,3-Diazole) Bound to Lys-41 in the Purple Membrane.- 5. Conformational Prediction of Bacteriorhodopsin Molecule.- 6. References.- 8 Structure and Dynamics of the Liver Microsomal Monoxygenase System.- 1. Introduction.- 1.1. General Structure of Biological Membranes.- 1.2. Peroxidation of Membrane Lipids.- 1.3. Microsomal Monoxygenase.- 2. Membrane Dynamics and Order Studied by Fluorescence.- 2.1. Biophysical Consequences of Lipid Peroxidation.- 2.2. Mobility of Membrane-Bound Cytochrome P-450.- 2.3. Rotational Mobility of Cytochrome P-450 in Peroxidized Rat Liver Microsomes.- 2.4. Structure of Free and Membrane-Bound Cytochrome P-450.- 2.5. Structure of NADPH-Cytochrome P-450 Reductase.- 2.6. Interaction of Cytochrome P-450 and Its Reductase in Membranes.- 2.7. Lipid-Protein Interactions Studied by DPH Fluorescence Anisotropy.- 3. References.- 9 Fluorescence Studies on Prokaryotic Membranes.- 1. Introduction.- 2. Fluorescent Probes.- 3. Structural Aspects of Bacterial Membranes.- 3.1. Outer Membrane of Gram-Negative Bacteria.- 3.2. Molecular Interactions.- 3.3. Phase Transitions and Homeoviscous Adaptation.- 3.4. Effects of Alcohols.- 3.5. Permeability of the Outer Membrane to Hydrophobic Substances.- 3.6. Membrane-Potential-Related Permeability Changes.- 3.7. Factors Increasing Cell Resistance and Membrane Stability.- 4. Periplasm.- 5. Incorporation of Exogenous Lipids into Prokaryotic Membranes.- 5.1. Gram-Negative Bacteria.- 5.2. Other Bacteria.- 5.3. Effect of Lipid Uptake on Membrane Function.- 5.4. Interactions with Vehicle Liposomes.- 6. Concluding Remarks.- 7. References.- 10 The Study of Cytoskeletal Protein Interactions by Fluorescence Probe Techniques.- 1. Introduction.- 2. The Cytoskeleton.- 2.1. Organization of Cytoskeletal Proteins.- 2.2. Assembly of Actin Filaments.- 3. Fluorescence Probe Techniques.- 3.1. Introduction.- 3.2. Energy Transfer.- 3.3. Fluorescence Enhancement.- 3.4. Anisotropy.- 3.5. Fluorescence Photobleaching Recovery.- 3.6. Quenching.- 3.7. Pressure Relaxation.- 4. Alternative Luminescence Techniques.- 4.1. Introduction.- 4.2. Transient Absorption Anisotropy.- 4.3. Phosphorescence.- 5. Summary and Future Prospects.- 6. References.- 11 Fluorescent Probes for the Acetylcholine Receptor Surface Environments.- 1. Introduction.- 2. An Overview of AchR Properties.- 2.1. Structural Characteristics of Torpedo californica AchR.- 2.2. Ligand Binding and Pharmacological Properties.- 3. PTSA: A Probe for Measuring AchR-Mediated Ionic Fluxes in the Physiological Time Scale.- 3.1. Stopped-Flow Assays for AchR-Mediated Ionic Fluxes.- 3.2. Stopped-Flow Assay with PTSA and Thallous Ion.- 4. Pyrene-1-Sulfonyl Azide (PySA): A Probe for the Study of the AchR-Lipid Interface.- 4.1. Optical Properties of PySA and Its Photoproducts.- 4.2. Labeling of the AchR.- 4.3. Applications of PySA to the Study of AchR Structure and Function.- 5. Pyrene Maleimide (PM): The Labeling of a Functionally Relevant Sulfhydryl Group.- 5.1. Labeling of Solubilized Receptor.- 5.2. Labeling of Native Membranes.- 6. State and Organization of the Lipid Bilayer in AchR Membranes.- 6.1. Probing of AchR Membranes with Pyrene.- 6.2. Fluidity of the AchR Membranes as Probed by DPH and TMA-DPH.- 7. Summary.- 8. References.- 12 Structural Basis and Physiological Control of Membrane Fluidity in Normal and Tumor Cells.- 1. Introduction.- 2. Quantitative Contribution of Individual Types of Lipid to Membrane Fluidity.- 2.1. Cholesterol, Sphingomyelin, and Fatty Acyl (Un)saturation.- 2.2. Cell Biological Implications of Preferential Interactions between Individual Lipids.- 3. Alterations in Membrane Fluidity in Lymphoid Tumor Cells.- 3.1. Tumor Cell Type and Location.- 3.2. Plasma Lipoproteins and Cholesterol Biosynthesis.- 4. Effects of Dietary Lipids on Membrane Fluidity.- 5. References.- 13 Fusion of Enveloped Viruses with Biological Membranes: Fluorescence Dequenching Studies.- 1. Introduction.- 2. Receptors for Enveloped Viruses.- 2.1. Myxoviruses.- 2.2. Paramyxoviruses.- 2.3. Togaviruses.- 2.4. Rhabdoviruses.- 2.5. Retroviruses.- 2.6. Herpesviruses.- 2.7. Other Enveloped Viruses.- 3. Interaction of Enveloped Viruses with Receptor-Depleted Cells.- 3.1. Use of Antimembrane Antibodies or Polypeptide Hormones to Mediate Virus Attachment.- 3.2. Implantation of Receptors or Binding Proteins for Enveloped Virions into Recipient Cell Membranes.- 4. Theoretical Aspects of the Use of Fluorescence Dequenching to Measure Viral Fusion.- 5. Fusion of Enveloped Viruses with Animal Cells and Biological Membranes: Studies with Intact Virions.- 6. Use of Fluorescent Dequenching Methods to Study Fusion of Enveloped Viruses with Biological Membranes Lacking Virus Receptors.- 7. Role of Viral Glycoproteins in the Process of Virus Membrane Fusion: Studies with Reconstituted Viral Envelopes.- 8. Fusion of Enveloped Viruses with Negatively Charged and Neutral Liposomes.- 9. Role of Conformational Changes and Cooperativity of Viral Proteins in Mediating Membrane Fusion.- 10. Conclusions.- 11. References.