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A ribosome inactivating protein from hairy melon (Benincasa hispida var. chieh-qua) seeds and peptides with translation-inhibiting activity from several other cucurbitaceous seeds.January 2001 (has links)
Parkash Amarender. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 158-172). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Table of contents --- p.ii / Abstract --- p.xi / 撮要 --- p.xiv / List of Abbreviations --- p.xvi / List of Tables --- p.xvii / List of Figures --- p.xix / Chapter CHAPTER 1. --- INTRODUCTION / Chapter 1.1 --- Ribosome-inactivating proteins (RIPs) --- p.3 / Chapter 1.2 --- General Properties of RIPs --- p.5 / Chapter 1.2.1 --- Structure --- p.5 / Chapter 1.2.1.1 --- Type I and Type II RIPs --- p.5 / Chapter 1.2.1.2 --- Small RIPs --- p.10 / Chapter 1.2.2 --- Distribution --- p.12 / Chapter 1.2.3 --- Physicochemical properties --- p.15 / Chapter 1.3 --- Enzymatic activities of RIPs --- p.17 / Chapter 1.3.1 --- N-glycosidase activity --- p.17 / Chapter 1.3.2 --- Polynucleotide:adenosine glycosidase activity --- p.21 / Chapter 1.3.3 --- Ribonuclease (RNase) activity --- p.24 / Chapter 1.3.4 --- Deoxyribonucleolytic (DNase) activity --- p.25 / Chapter 1.3.5 --- Multiple depurination --- p.26 / Chapter 1.3.6 --- Inhibition of protein synthesis --- p.27 / Chapter 1.4 --- Biological activities of RIPs --- p.29 / Chapter 1.4.1 --- Interaction of ribosome-inactivating proteins with cells --- p.29 / Chapter 1.4.1.1 --- Internalization of type 1 ribosome-inactivating proteins --- p.29 / Chapter 1.4.1.2 --- Internalization of type 2 ribosome-inactivating proteins --- p.32 / Chapter 1.4.2 --- Effects on laboratory animals --- p.33 / Chapter 1.4.3 --- Immunosuppressive activity --- p.33 / Chapter 1.4.4 --- Abortifacient activity --- p.34 / Chapter 1.4.5 --- Antiviral activity --- p.35 / Chapter 1.5 --- Physiological roles of RIPs --- p.37 / Chapter 1.6 --- Applications of RIPs --- p.39 / Chapter 1.6.1 --- Possible uses in experimental and clinical medicine --- p.39 / Chapter 1.6.1.1 --- Anti-tumor therapy --- p.40 / Chapter 1.6.1.2 --- Immune disorders --- p.42 / Chapter 1.6.1.3 --- Neuroscience research --- p.43 / Chapter 1.6.2 --- Applications in agriculture --- p.44 / Chapter 1.7 --- Arginine/Glutamate Rich Polypeptides (AGRPs) --- p.46 / Chapter 1.8 --- Objectives of the present study --- p.48 / Chapter 1.8.1 --- Rationale of the study --- p.48 / Chapter 1.8.2 --- Outline of the thesis --- p.50 / Chapter Chapter 2 --- Materials and methods / Chapter 2.1 --- Introduction --- p.52 / Chapter 2.2 --- Materials and methods --- p.54 / Chapter 2.2.1 --- Materials --- p.54 / Chapter 2.2.2 --- Preparation of crude extract --- p.55 / Chapter 2.2.3 --- Purification of proteins --- p.55 / Chapter 2.2.4 --- Molecular weight determination with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) --- p.61 / Chapter 2.2.5 --- Protein determination --- p.64 / Chapter 2.2.6 --- N-terminal amino acid sequence --- p.64 / Chapter 2.2.7 --- Preparation of rabbit reticulocyte lysate --- p.65 / Chapter 2.2.8 --- Assay for cell-free protein synthesis- inhibiting activity --- p.65 / Chapter 2.2.9 --- Assay for N-glycosidase activity --- p.66 / Chapter 2.2.10 --- Assay for ribonuclease activity --- p.70 / Chapter 2.2.11 --- Assay for antifungal activity --- p.71 / Chapter 2.2.12 --- Assay for dehydrogenase activity --- p.71 / Chapter Chapter 3 --- Purification and characterization of proteins from their respective sources. / Chapter 3.1. --- Purification and Characterization of Hispidin from Hairy melon (Benincasa hispida var. chieh-qua) / Chapter 3.1.1. --- Introduction --- p.73 / Chapter 3.1.2. --- Results --- p.76 / Chapter 3.1.2.1. --- Purification --- p.78 / Chapter 3.1.2.2. --- Molecular weight determination --- p.84 / Chapter 3.1.2.3. --- N-terminal amino acid sequence --- p.85 / Chapter 3.1.2.4. --- Assay for cell-free protein synthesis-inhibiting activity --- p.86 / Chapter 3.1.2.5. --- Assay for N-glycosidase activity --- p.87 / Chapter 3.1.2.6. --- Assay for ribonuclease activity --- p.88 / Chapter 3.1.2.7. --- Assay for dihydrodiol dehydrogenase activity --- p.88 / Chapter 3.1.2.8. --- Assay for antifungal activity --- p.89 / Chapter 3.1.2.9. --- "Assessment of purity, yield and activity" --- p.91 / Chapter 3.1.3. --- Discussion --- p.92 / Chapter 3.2. --- Purification and Characterization of Momorchin from Dried Bitter Gourd (Momordica charantia) Seeds / Chapter 3.2.1. --- Introduction --- p.95 / Chapter 3.2.2. --- Results --- p.99 / Chapter 3.2.2.1. --- Purification --- p.100 / Chapter 3.2.2.2. --- Molecular weight determination --- p.103 / Chapter 3.2.2.3. --- N-terminal amino acid sequence --- p.104 / Chapter 3.2.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.105 / Chapter 3.2.2.5. --- Assay for ribonuclease activity --- p.105 / Chapter 3.2.2.6. --- Assay for N-glycosidase activity --- p.106 / Chapter 3.2.2.7. --- "Assessment of purity, yield and activity" --- p.107 / Chapter 3.2.3. --- Discussion --- p.108 / Chapter 3.3.3. --- Purification and Characterization of Luffacylin from Sponge Gourd (Luffa cylindrica) / Chapter 3.3.1. --- Introduction --- p.110 / Chapter 3.3.2. --- Results --- p.113 / Chapter 3.3.2.1. --- Purification --- p.115 / Chapter 3.3.2.2. --- Molecular weight determination --- p.119 / Chapter 3.3.2.3. --- N-terminal amino acid sequencing --- p.120 / Chapter 3.3.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.121 / Chapter 3.3.2.5. --- Assay for ribonuclease activity --- p.121 / Chapter 3.3.2.6. --- Assay for N-glycosidase activity --- p.122 / Chapter 3.3.2.7. --- Assay for antifungal activity --- p.123 / Chapter 3.3.2.8. --- "Assessment of purity, activity and yield" --- p.124 / Chapter 3.3.3. --- Discussion --- p.125 / Chapter 3.4. --- Purification and Characterization of α and β Benincasin from fresh Winter Melon {Benincasa hispida var. dong-gua) Seeds / Chapter 3.4.1. --- Introduction --- p.127 / Chapter 3.4.2. --- Results --- p.129 / Chapter 3.4.2.1. --- Purification --- p.130 / Chapter 3.4.2.2. --- Molecular weight determination --- p.135 / Chapter 3.4.2.3. --- N-terminal amino acid sequence --- p.136 / Chapter 3.4.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.137 / Chapter 3.4.2.5. --- Assay for ribonuclease activity --- p.137 / Chapter 3.4.2.6. --- Assay for antifungal activity --- p.138 / Chapter 3.4.2.7. --- "Assessment of purity, activity and yield" --- p.140 / Chapter 3.4.3. --- Discussion --- p.141 / Chapter 3.5. --- Purification and characterization of Moschins from Pumpkin (Cucurbita moschata) Seeds / Chapter 3.5.1. --- Introduction --- p.143 / Chapter 3.5.2. --- Results --- p.145 / Chapter 3.5.2.1. --- Purification --- p.146 / Chapter 3.5.2.2. --- Molecular weight determination --- p.149 / Chapter 3.5.2.3. --- N-terminal amino acid sequence --- p.150 / Chapter 3.5.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.151 / Chapter 3.5.2.5. --- Assay for ribonuclease activity --- p.151 / Chapter 3.5.2.6. --- "Assessment of purity, activity and yield" --- p.152 / Chapter 3.5.3. --- Discussion --- p.153 / Chapter Chapter 4 --- General Discussion and Conclusion --- p.154 / References --- p.158
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