000			04481nam a2200145Ia 4500
999			_c176467 _d176467
020			_a9788182930452
040			_cCUS
082			_a612 _bSIN/P
100			_aSingh, Ram Dayal _921706
245		0	_aProtein Microarrays/ _c Ram Dayal Singh
260			_aNew Delhi: _bInternational Scientific Publishing Academy, _c2011.
300			_ax, 278p.
505			_a 1. PROTEIN MICROARRAYS 1 Protein Therapeutics 2 Applications of Protein Microarrays 3 Problems and Challenges 5 Sample Preparation and Handling 5 Microarray Platform 7 Detection Technologies 7 Data Analysis 8 Potential Solutions 8 2. DEVELOPMENT OF A PROTEIN MICROARRAY 11 Protein Microarray Technologies 13 Assays on a Microspot 15 Types of Spot-based Array Formats 16 Capture Agents 19 Solid Support and Surface Chemistry 21 Arraying Devices for Immobilisation of Capture Agents 23 Signal Generation and Signal Detection 24 Classification Protein Microarrays 27 Protein Expression Microarrays 27 Protein Function Microarrays 29 Protein Biomarker Screening 30 Rheumatoid Arthritis 32 Etiology of- Rheumatoid Arthritis 34 Pathogenesis of Rheumatoid Arthritis 35 Treatment of Rheumatoid Arthritis 39VIII Protein Microarra'ss 3. PROTEIN MICROARRAY GENERATION AND DETECTION 42 Immobilization Strategies 43 Via Biotin-Avidin Interaction 44 In Vitro Biotinylation of Proteins 44 PCR-based In vitro Biotinylation of Proteins 47 In vivo Biotinylation of Proteins 48 Via N-terminal Cysteine 49 Detection Strategies 50 Mechanism-based Inhibitor Probes on Array 52 Protein Profiling using Fluorogenic Substrates 53 Array Fabrication 56 4. PROTEIN STRUCTURE 58 Peptide Bond 61 Protein Secondary Structure 62 a-Helix 62 Effects of Amino Acid Sequence 65 p Conformation 68 Characteristic Bond Angles and Amino Acid Contents 69 Protein Tertiary and Quaternary Structures 71 Fibrous Proteins 72 a-Keratin 73 Functional Diversity in Globular Proteins 79 Tertiary Structures in Globular Proteins 83 Common Structural Patterns 86 Structural Classification 91 Protein Quaternary Structures 92 5. PROPERTIES AND EVOLUTION OF PROTEINS 97 Electrophoresis 100 Unseparated Proteins 104 Covalent Structure of Proteins 106 Role ofAmino Acid Sequence 107 Sequenced using Automated Procedures 109 Sequence of Large Proteins 111 Breaking Disulfide Bonds III Sequencing of Peptides 112 Ordering Peptide Fragments US Locating Disulfide Bonds 113CONTKNTS IX Small Peptide and Proteins 114 Protein Sequences and Evolution 117 6. PROTEIN DENATURATION AND FOLDING 125 Amino Acid Sequence Determines Tertiary Structure 127 Polypeptides Fold Rapidly by a Stepwise Process 128 Protein Folding 132 Principles of Protein Folding 134 Hydrophobic Interactions 135 Experimental Evidence 136 Simple Exact Models 137 Salt-Induced Detour 141 Triangular Scheme between D,C and N 143 Using Salt to Collapse the Denatured Ensemble 145 Protein Engineering Analysis of the Structure ofC 149 7. FOLDING FUNNELS AND FRUSTRATION 154 p-Barrel Model 1^2 Signatures of Good and Bad Folders 164 Thermodynamics 154 Kinetics 155 8. PROTEIN FOLDING PATHWAYS AND KINETICS 170 Characteristics of Folding Kinetics 17g Intermediates and Folding Pathways 184 Folding Speed and Topology 187 Kinetic Folding Pathway of p-sheet Protein 191 Stopped-flow Fluorescence Measurements 195 Quenched-flow Experiments 195 Equilibrium Unfolding 198 Anomalous CD Overshoot Phenomenon 200 Amide Proton Protection 202 Hydrophobic Collapse Detected by ANS Fluorescence 204 Comparison with Folding Kinetics ofInterleukin-1p 208 Significance ofthe Hydrophobic Collapse 209 9. PROTEIN DESIGN 212 Protein Modeling 214 Learning the Interaction Potentials 216 Designing PDB Structures 220 Similarities of Homologous Sequences 222 10. evolution OFPROTEIN THERMODYNAMICS 229 Evolutionary Steady-state 231X Protein Microarrans Protein Sequence-structure Compatibility 240 Pfotein Structure Prediction and Sequence Design 242 11. CANCER CELLS USING PROTEIN MICROARRAYS 244 Cell—Lines Culture,and Irradiation 246 Extraction and Delipidation 246 Detection of Protein Interactions 248 Analysis of Differential Protein Expression 248 Imunoblot Analysis 250 Qualitative Protein Levels in Antibody Microarrays 250 12. VALIDATION OF MICROARRAYS 260 AimsofValidation 260 Challenges ofValidation 261 Validation in Silico 264 Validation using PCR 268 Qualitative PCR 269 Semi-Quantitative PCR 270 Real-time Quantitative PCR 273 Double-stranded DNA-binding Agents 273 Validation Using Protein Antibodies 277
942			_cWB16 _01