Photonics of biopolymers /
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ISBN：9783540438175
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简介
Photonics of biopolymers discusses the processes of energy transformation in photoexcited proteins, nucleic acids, membranes and model systems. The author employs his considerable experience to deliver a thorough explanation of the physical background of the optical techniques for studying biological systems, and he identifies problems and experimental pitfalls that prevent accurate measurements and interpretation. He not only describes the spectroscopic methods, he uses his own experimental results to extract information from the data and critically evaluates the competing models used to interpret the results.
目录
Introduction to Photonics p. 1
Light Absorption p. 3
Vibrational Relaxation and Internal Conversion p. 3
Fluorescence p. 3
Intersystem Crossing and Phosphorescence p. 4
Energy Transfer p. 5
Excimer and Exciplex Formation p. 5
Photochemical Reaction and Electron Transfer p. 5
Photoinduced Conformational Changes p. 5
Light Absorption in Ordered Structures p. 7
Probability of Absorption p. 7
Lambert-Beer Law p. 9
Biological Chromophores p. 10
Absorbance in Scattering Media p. 11
Hypochromism p. 11
Molecular Interactions p. 12
Sieve Effect p. 13
Light-Scattering Model p. 14
Light-Dispersion Model p. 15
Stacking Model p. 15
Conclusion p. 16
Screening Hypochromism p. 18
Screening model p. 19
Nucleotides, Oligonucleotides and DNA p. 23
Tyrosine and Tryptophan in Aggregates p. 27
Tyrosine and Tryptophan in Proteins p. 28
Hemoglobin in Erythrocytes p. 30
Chloroplasts and Visual Rods p. 31
Clusters of Aromatic Hydrocarbons p. 32
Conclusion p. 35
Photometric Estimation of Protein Content in Biological Suspensions p. 36
Colorimetry and Others Methods p. 36
Combined UV-Spectrophotometric Method p. 36
Comparison of the Methods p. 38
Conclusion p. 40
Screening and Reabsorption of Light p. 41
Inner Filter Effect p. 41
Microscreening and Microreabsorption p. 42
Erythrocytes and Other Cells p. 44
Volume Reabsorption of Donor Luminescence p. 45
Trypaflavine Activates Rhodamine Fluorescence p. 47
Conclusion p. 50
Multipass Cuvettes for Luminescence Spectroscopy p. 51
Mirror and Total Internal Reflection Cuvettes p. 51
Multipass Cuvettes in Steady-State Measurements p. 53
Multipass Cuvettes in Lifetime Measurements p. 54
Other Applications p. 55
Conclusion p. 55
Division of Tyrosine and Tryptophan Fluorescence Components p. 56
Generally Accepted Approaches p. 56
Synchronous Scanning Method p. 58
Synchronous Spectra of Tryptophan and Tyrosine p. 58
Synchronous Spectra of Different Tryptophan Residues p. 60
Conclusion p. 61
Spectral Heterogeneity of Tryptophan Emission p. 62
Variation of Fluorescence Polarization along Tryptophan Emission Spectrum p. 62
Fluorescence Lifetime Variations p. 68
Photoinduced Conformational Mobility of Proteins p. 70
Phosphorescence of Proteins p. 72
Conclusion p. 73
Discrete Emission States in Photoexcited Tryptophan Complexes p. 74
Time-Resolved Spectroscopy of Tryptophan Fluorescence p. 75
TRP and NATA in Water p. 78
TRP, NATA and Indole in Ethanol p. 80
TRP and NATA in Glycerol p. 81
Dipeptides p. 83
Exciplexes in Proteins p. 84
Conclusion p. 88
Mechanisms of Exciplex Formation p. 90
Generally Accepted Models p. 90
Fractional Energy Transfer in Exciplexes p. 92
Exciplex of Aromatic Hydrocarbons p. 94
Excimers p. 96
Pyrene-Indole Exciplex p. 98
Conclusion p. 100
Mechanisms of Energy Transfer p. 101
Inductive-Resonance Model p. 102
Energy Transfer in Molecular Structures p. 105
Hot Migration p. 108
Conclusion p. 110
Energy Transfer in Nucleic Acids p. 111
Migration Between Nucleotides p. 111
Migration Along DNA p. 113
Quantum Yield of Energy Transfer to Dyes p. 113
Polyadenilic Acid Labeled by Ethenoadenine p. 115
DNA with Intercalated Dyes p. 115
Fluorescent Probes and Labels on DNA p. 119
Conclusion p. 121
Energy Transfer in Native Proteins p. 122
Tyrosine-Tryptophan Pair p. 122
Migration between Tryptophan Residues p. 126
Tryptophan-NADH Pair in Alcohol Dehydrogenase p. 127
Tryptophan-Heme Pair in Myoglobin p. 132
Tryptophan-Pyrene Pair p. 134
Quenching of Tryptophan Emission by Dyes p. 137
Conclusion p. 141
Energy Transfer in Biomembranes p. 142
Quenching of Tryptophan Fluorescence in Sarcoplasmic Reticulum by Probes p. 143
Quenching of Tryptophan Fluorescence by ANS p. 145
Quenching of Tryptophan Fluorescence by Pyrene p. 146
Tryptophan-NADH Pair in Mitochondria p. 147
Photosynthetic Reaction Centers p. 149
Conclusion p. 150
Fluorescence Probes p. 151
Widely Used Probes p. 151
Estimation of Sizes of Chaperones and their Complexes Using ANS p. 155
Fluorescent Studies of Na[superscript +]K[superscript +]-ATPase p. 158
Anthracene with Dimethylaminochalcone in Membranes p. 159
Diffusion of Probes p. 161
Fluorescence Pharmacology in vitro p. 162
Conclusion p. 163
Pyrene Monomers and Excimers in Membranes p. 165
Viscosity Measurements p. 165
Location and Diffusion of Pyrene p. 166
Detection of Oxygen by Pyrene Emission p. 168
Vibronic Peaks as Indicators of Membrane Polarity p. 170
Conclusion p. 171
Photomodulation of Enzyme Activity p. 172
Photoactivation of Enzymes p. 172
Photodesorption p. 173
Photochemical Processes in Alcohol Dehydrogenase p. 174
Photolysis of Flavin in NADH Dehydrogenase p. 179
Conclusion p. 183
Photoactivation of Animal Membranes and their Chromophores p. 184
Photoinduced Membranes Activity p. 184
Oxygen Uptake in Mitochondria under Photoexcitation p. 185
Oxidation of NADH by Triplet Flavin and Singlet Oxygen p. 187
Conclusion p. 192
Light-Dependent Phosphorylation in Mitochondria p. 193
ATP Synthesis during Illumination p. 193
Thermal Coupling between ATP Synthesis and Electron Transfer p. 197
Conclusion p. 199
References p. 200
Index p. 228
Light Absorption p. 3
Vibrational Relaxation and Internal Conversion p. 3
Fluorescence p. 3
Intersystem Crossing and Phosphorescence p. 4
Energy Transfer p. 5
Excimer and Exciplex Formation p. 5
Photochemical Reaction and Electron Transfer p. 5
Photoinduced Conformational Changes p. 5
Light Absorption in Ordered Structures p. 7
Probability of Absorption p. 7
Lambert-Beer Law p. 9
Biological Chromophores p. 10
Absorbance in Scattering Media p. 11
Hypochromism p. 11
Molecular Interactions p. 12
Sieve Effect p. 13
Light-Scattering Model p. 14
Light-Dispersion Model p. 15
Stacking Model p. 15
Conclusion p. 16
Screening Hypochromism p. 18
Screening model p. 19
Nucleotides, Oligonucleotides and DNA p. 23
Tyrosine and Tryptophan in Aggregates p. 27
Tyrosine and Tryptophan in Proteins p. 28
Hemoglobin in Erythrocytes p. 30
Chloroplasts and Visual Rods p. 31
Clusters of Aromatic Hydrocarbons p. 32
Conclusion p. 35
Photometric Estimation of Protein Content in Biological Suspensions p. 36
Colorimetry and Others Methods p. 36
Combined UV-Spectrophotometric Method p. 36
Comparison of the Methods p. 38
Conclusion p. 40
Screening and Reabsorption of Light p. 41
Inner Filter Effect p. 41
Microscreening and Microreabsorption p. 42
Erythrocytes and Other Cells p. 44
Volume Reabsorption of Donor Luminescence p. 45
Trypaflavine Activates Rhodamine Fluorescence p. 47
Conclusion p. 50
Multipass Cuvettes for Luminescence Spectroscopy p. 51
Mirror and Total Internal Reflection Cuvettes p. 51
Multipass Cuvettes in Steady-State Measurements p. 53
Multipass Cuvettes in Lifetime Measurements p. 54
Other Applications p. 55
Conclusion p. 55
Division of Tyrosine and Tryptophan Fluorescence Components p. 56
Generally Accepted Approaches p. 56
Synchronous Scanning Method p. 58
Synchronous Spectra of Tryptophan and Tyrosine p. 58
Synchronous Spectra of Different Tryptophan Residues p. 60
Conclusion p. 61
Spectral Heterogeneity of Tryptophan Emission p. 62
Variation of Fluorescence Polarization along Tryptophan Emission Spectrum p. 62
Fluorescence Lifetime Variations p. 68
Photoinduced Conformational Mobility of Proteins p. 70
Phosphorescence of Proteins p. 72
Conclusion p. 73
Discrete Emission States in Photoexcited Tryptophan Complexes p. 74
Time-Resolved Spectroscopy of Tryptophan Fluorescence p. 75
TRP and NATA in Water p. 78
TRP, NATA and Indole in Ethanol p. 80
TRP and NATA in Glycerol p. 81
Dipeptides p. 83
Exciplexes in Proteins p. 84
Conclusion p. 88
Mechanisms of Exciplex Formation p. 90
Generally Accepted Models p. 90
Fractional Energy Transfer in Exciplexes p. 92
Exciplex of Aromatic Hydrocarbons p. 94
Excimers p. 96
Pyrene-Indole Exciplex p. 98
Conclusion p. 100
Mechanisms of Energy Transfer p. 101
Inductive-Resonance Model p. 102
Energy Transfer in Molecular Structures p. 105
Hot Migration p. 108
Conclusion p. 110
Energy Transfer in Nucleic Acids p. 111
Migration Between Nucleotides p. 111
Migration Along DNA p. 113
Quantum Yield of Energy Transfer to Dyes p. 113
Polyadenilic Acid Labeled by Ethenoadenine p. 115
DNA with Intercalated Dyes p. 115
Fluorescent Probes and Labels on DNA p. 119
Conclusion p. 121
Energy Transfer in Native Proteins p. 122
Tyrosine-Tryptophan Pair p. 122
Migration between Tryptophan Residues p. 126
Tryptophan-NADH Pair in Alcohol Dehydrogenase p. 127
Tryptophan-Heme Pair in Myoglobin p. 132
Tryptophan-Pyrene Pair p. 134
Quenching of Tryptophan Emission by Dyes p. 137
Conclusion p. 141
Energy Transfer in Biomembranes p. 142
Quenching of Tryptophan Fluorescence in Sarcoplasmic Reticulum by Probes p. 143
Quenching of Tryptophan Fluorescence by ANS p. 145
Quenching of Tryptophan Fluorescence by Pyrene p. 146
Tryptophan-NADH Pair in Mitochondria p. 147
Photosynthetic Reaction Centers p. 149
Conclusion p. 150
Fluorescence Probes p. 151
Widely Used Probes p. 151
Estimation of Sizes of Chaperones and their Complexes Using ANS p. 155
Fluorescent Studies of Na[superscript +]K[superscript +]-ATPase p. 158
Anthracene with Dimethylaminochalcone in Membranes p. 159
Diffusion of Probes p. 161
Fluorescence Pharmacology in vitro p. 162
Conclusion p. 163
Pyrene Monomers and Excimers in Membranes p. 165
Viscosity Measurements p. 165
Location and Diffusion of Pyrene p. 166
Detection of Oxygen by Pyrene Emission p. 168
Vibronic Peaks as Indicators of Membrane Polarity p. 170
Conclusion p. 171
Photomodulation of Enzyme Activity p. 172
Photoactivation of Enzymes p. 172
Photodesorption p. 173
Photochemical Processes in Alcohol Dehydrogenase p. 174
Photolysis of Flavin in NADH Dehydrogenase p. 179
Conclusion p. 183
Photoactivation of Animal Membranes and their Chromophores p. 184
Photoinduced Membranes Activity p. 184
Oxygen Uptake in Mitochondria under Photoexcitation p. 185
Oxidation of NADH by Triplet Flavin and Singlet Oxygen p. 187
Conclusion p. 192
Light-Dependent Phosphorylation in Mitochondria p. 193
ATP Synthesis during Illumination p. 193
Thermal Coupling between ATP Synthesis and Electron Transfer p. 197
Conclusion p. 199
References p. 200
Index p. 228
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