Supramolecular Structure and Function 10 (eBook)

Preface 5
Contents 7
Contributors 8
Fluorescence Correlation Spectroscopy: Principles and Developments 10
1 Introduction 11
2 Principles of FCS 12
2.1 What Is FCS? 12
2.1.1 Fluorescence 12
2.1.2 Correlation 14
2.1.3 Spectroscopy 15
2.2 The Geometrical Factor 16
2.3 The Signal-to-Noise in FCS 18
2.4 The Limitations of FCS 19
2.5 Reactions 20
2.5.1 Triplet-State 20
2.5.2 Unimolecular Reactions 21
2.5.3 Bimolecular Reactions 21
2.6 Multiple Species 23
2.7 Cross-Correlation Spectroscopy 25
2.8 Pulsed Interleaved Excitation 25
3 Advanced FCS Methods 30
3.1 Scanning FCS 30
3.2 Two-Focus FCS 33
3.3 Raster Image Correlation Spectroscopy 35
References 36
Time-Resolved FT-IR Spectroscopy for the Elucidation of Protein Function 40
1 Introduction 40
2 Time-Resolved FT-IR Techniques 41
2.1 Rapid-Scan FT-IR Spectroscopy 41
2.2 Step-Scan FT-IR Spectroscopy 42
2.2.1 Step-Scan Device 43
2.2.2 Step-Scan Timing 44
2.2.3 Step-Scan Parameters 44
3 Applications: Bacteriorhodopsin 44
4 New Developments of the Step-Scan FT-IR Technique 47
References 48
Recombinant Membrane Protein Production: Past, Present and Future 50
1 Introduction 50
2 Membrane Targeting and Insertion 51
3 Naturally Abundant Membrane Proteins 54
4 Expression Systems 55
4.1 Prokaryotic Expression Systems 55
4.1.1 Escherichia Coli 55
4.1.2 Lactococcus Lactis 56
4.1.3 Alternative Bacterial Expression Systems 60
4.2 Eukaryotic Expression Systems 60
4.2.1 Yeasts 61
4.2.2 Insect Cells 62
4.2.3 Mammalian Cells 63
4.2.4 Alternative Expression Systems 63
4.3 Cell-Free Expression 64
5 Expression Tools 65
6 Host Screening/Strain Selection 66
7 Protein Engineering 67
8 Homologue Screening 68
9 Stress Response Upon Membrane Protein Expression 68
10 Rational Engineering of Membrane Protein Expression Through Co-expression of Limiting Factors 70
11 Strategies to Optimize Functional Expression 71
12 Conclusions 73
References 74
Cold Denaturation and Protein Stability 84
1 Introduction 85
2 Unbiased Cold Denaturation 86
3 Alcohols: Denaturants or Structure Stabilizers? 88
4 Stability and Aggregation 91
References 93
Polyglutamine Diseases and Neurodegeneration: The Example of Ataxin-1 95
1 Introduction 95
2 Structural Features of Atx1 96
2.1 Atx1 Domain Architecture 96
2.2 The Structure of the Atx1 AXH Domain 97
2.3 Atx1 ULM Functions as a Molecular Switch 98
3 Atx1 Functions 98
3.1 A Role for Atx1 in Transcriptional Repression 98
3.2 RNA Processing 101
3.3 The Multiple Role of the AXH Motif in Nucleic Acid- and Protein-Protein Interactions 102
3.4 Towards the Determination of the Atx1 Interactome 103
4 Proteasome Functions in SCA1 103
5 Conclusions 104
References 105
Phase Plate Electron Microscopy 108
1 Introduction 108
1.1 Defocus Phase Contrast TEM 109
1.2 Zernike Phase Contrast TEM 109
1.3 Hilbert Differential Contrast TEM 111
1.4 Phase Contrast Cryo-TEM 111
2 Biological Applications to Two Dimensional Imaging 113
2.1 ZPC-TEM Applications in Two Dimensions 113
2.2 HDC-TEM Applications in Two Dimensions 115
3 Biological Applications to Three Dimensional Imaging 116
3.1 ZPC-TEM Applications in Three Dimensions 116
4 Future Prospects and Conclusions 116
4.1 Electrostatic Charging 116
4.2 Next-Generation Phase Plates 118
References 118
Deriving Biomedical Diagnostics from Spectroscopic Data 121
1 Introduction 121
2 The Problems 122
3 Principal Component Analysis and Simca 124
4 Partial Least Squares (PLS) and Principal Component Regression (PCR) 124
5 The Statistical Classification Strategy 125
6 Examples 127
6.1 Screening for Colon Cancer 127
6.2 Thyroid Cancer 128
6.3 Biliary Cancer 128
7 The Future 129
References 129
The Emergence and Ozone Treatment Studies of Living Cells 131
1 Introduction 131
2 On High-Frequency Bio Oxidative Treatments (HF-BOT) 132
3 Some Examples of Dental and Medical Effects of HFBot 136
References 137
Toxicity Study of Nanofibers 139
1 Introduction 139
1.1 Carbon Nanotubes 140
1.2 Inorganic Nanofilaments 140
1.3 Boron Nitride Nanotubes 141
2 Current State of the Reserach 141
2.1 Potential Toxicity of CNTs 142
2.2 Potential Toxicity of INFs 142
2.3 Potential Toxicity of BNNTs 143
3 Cellular Toxicity 143
3.1 Effect of Surface Chemistry 143
3.1.1 Materials and Methods 143
3.2 Effect of Morphology and Surface Chemistry 145
3.2.1 Materials and Methods 146
3.3 Possible Effect of Tortuosity 149
3.3.1 Materials and Methods 150
4 Conclusions and Future Perspectives 152
References 153
Subject Index 156