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Isolation and characterization of five pathogenesis-related proteins from Panax notoginseng, Lyophyllum shimeji and Hypsizigus marmoreus.

Lam Sze Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 172-200). / Abstracts in English and Chinese. / Acknowledgements --- p.ii / Table of contents --- p.ii / Abstract --- p.xii / 撮要 --- p.xv / List of Abbreviations --- p.xvi / List of Tables --- p.xvii / List of Figures --- p.xix / TABLE OF CONTENTS / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1. --- Overview of Chitinases --- p.3 / Chapter 1.1.1. --- Classification of Chitinases --- p.7 / Chapter 1.1.1.1. --- Family 19 Chitinases --- p.7 / Chapter 1.1.1.1.1. --- Class I Chitinases --- p.9 / Chapter 1.1.1.1.2. --- Class II Chitinases --- p.10 / Chapter 1.1.1.1.3. --- Class IV Chitinases --- p.10 / Chapter 1.1.1.1.4. --- Class V Chitinases --- p.11 / Chapter 1.1.1.1.5. --- Class VI Chitinases --- p.11 / Chapter 1.1.1.2. --- Family 18 Chitinases --- p.12 / Chapter 1.1.1.2.1. --- PR-8/Class III Chitinases --- p.12 / Chapter 1.1.1.2.2. --- PR-11 Chitinases --- p.15 / Chapter 1.1.1.3. --- The PR-4 Family --- p.16 / Chapter 1.1.2. --- Catalytic Mechanism of Chitinases --- p.19 / Chapter 1.1.2.1. --- Catalytic Mechanism of Family 18 Chitinases --- p.20 / Chapter 1.1.2.2. --- Catalytic Mechanism of Family 19 Chitinases --- p.21 / Chapter 1.1.3. --- Biological Properties of Chitinases --- p.22 / Chapter 1.1.3.1. --- Antifungal Activity of Chitinases in vitro --- p.22 / Chapter 1.1.3.2. --- Antifungal Activity of Chitinases in vivo --- p.23 / Chapter 1.1.3.3. --- Other Functions --- p.23 / Chapter 1.2. --- Overview of Ribonucleases --- p.25 / Chapter 1.2.1. --- Classification of Ribonucleases --- p.26 / Chapter 1.2.1.1. --- RNase T1 Family --- p.26 / Chapter 1.2.1.1.1. --- Action Mechanism of RNase T1 Family --- p.32 / Chapter 1.2.1.2. --- RNase T2 Family --- p.34 / Chapter 1.2.1.2.1. --- Action Mechanism of RNase T2 Family --- p.36 / Chapter 1.2.2. --- Biological Activities of Plant Ribonucleases --- p.38 / Chapter 1.2.2.1. --- Phosphate Remobilization --- p.38 / Chapter 1.2.2.2. --- Senescence --- p.39 / Chapter 1.2.2.3. --- Programmed Cell Death --- p.40 / Chapter 1.2.2.4. --- Plant Defense --- p.41 / Chapter 1.2.2.5. --- RNA Processing and Decay --- p.43 / Chapter 1.2.2.6. --- Antitumor Activities --- p.43 / Chapter 1.3. --- Overview of plant ribosome-inactivating proteins (RIPs) --- p.45 / Chapter 1.3.1. --- General properties of RIPs --- p.46 / Chapter 1.3.1.1. --- Classification of RIPs --- p.46 / Chapter 1.3.2. --- Activities of Ribosome-inactivating Proteins --- p.52 / Chapter 1.3.2.1. --- RNA N-glycosidase activity --- p.52 / Chapter 1.3.2.2. --- Protein synthesis inhibitory activity --- p.58 / Chapter 1.3.2.3. --- Abortifacient activity --- p.59 / Chapter 1.3.2.4. --- Immunosuppressive activity --- p.60 / Chapter 1.3.2.5. --- Antiviral activity --- p.61 / Chapter 1.3.3. --- Roles of RIPs in plants --- p.63 / Chapter 1.3.3.1. --- Defensive role of RIPs in plants --- p.63 / Chapter 1.3.3.2. --- Role of RIPs in stress adaptation in plants --- p.66 / Chapter 1.3.4. --- Possible application of RIPs --- p.67 / Chapter 1.3.4.1. --- Use of RIPs in therapies --- p.67 / Chapter 1.3.4.1.1. --- Antiviral agents --- p.67 / Chapter 1.3.4.1.2. --- Immunotoxins --- p.68 / Chapter 1.3.4.1.3. --- Anti-HIV drugs --- p.69 / Chapter 1.3.4.2. --- Use of RIPs in agriculture --- p.71 / Chapter 1.4. --- Overview of the PR-5 Family: Thaumatin-Like Proteins (TLPs) --- p.72 / Chapter 1.4.1. --- Occurrence of Thaumatin-Like Proteins --- p.76 / Chapter 1.4.2. --- Biological properties of TLPs --- p.77 / Chapter 1.4.2.1. --- Antifungal Activity --- p.77 / Chapter 1.4.2.2. --- TLPs as Anti-Freeze Protein --- p.78 / Chapter 1.4.3. --- Biotechnological Application ´ؤ Transgenic Plants --- p.79 / Chapter Chapter 2 --- Materials and Methods --- p.81 / Chapter 2.1. --- Materials --- p.81 / Chapter 2.2. --- Preparation of Crude Extract --- p.82 / Chapter 2.3. --- Purification --- p.83 / Chapter 2.4. --- Chromatography --- p.84 / Chapter 2.4.1. --- CM-Cellulose Chromatography --- p.84 / Chapter 2.4.2. --- Mono S® HR 5/5 and Mono Q® HR 5/5 --- p.85 / Chapter 2.4.3. --- Affi-gel Blue gel --- p.86 / Chapter 2.4.4. --- Superdex75 --- p.87 / Chapter 2.5. --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.88 / Chapter 2.6. --- Protein Concentration Determination --- p.89 / Chapter 2.7. --- Preparation of Rabbit Reticulocyte Lysate --- p.90 / Chapter 2.8. --- Determination of N-terminal Amino Acid Sequence --- p.91 / Chapter 2.9. --- Biological Activity Assays --- p.92 / Chapter 2.9.1. --- Assay for Antifungal Activity --- p.92 / Chapter 2.9.2. --- Assay for Cell-Free Translation Inhibitory Activity --- p.93 / Chapter 2.9.3. --- Assay of Cytotoxic Activity on Cancer Cell Lines --- p.94 / Chapter 2.9.4. --- Assay for HIV-1 Reverse Transcriptase (RT) Inhibitory Activity --- p.95 / Chapter 2.9.5. --- Assay of Mitogenic Activity --- p.97 / Chapter 2.9.6. --- Assay for N-Glycosidase Activity --- p.98 / Chapter 2.9.6.1. --- RNA Extraction --- p.98 / Chapter 2.9.6.2. --- Aniline Treatment --- p.99 / Chapter 2.9.6.3. --- Formaldehyde Gel Electrophoresis --- p.99 / Chapter 2.9.7. --- Assay of Ribonuclease Activity --- p.100 / Chapter 2.9.7.1. --- Assay for Yeast tRNA --- p.100 / Chapter 2.9.7.2. --- Activity toward Polyhomoribonucleotides --- p.100 / Chapter Chapter 3 --- Purification and Characterization of Pathogenesis-Related Proteins from their Respective Sources --- p.101 / Chapter 3.1. --- Purification and Characterization of Chitinase and Ribonuclease from the Roots of Panax notoginseng --- p.102 / Chapter 3.1.1. --- Introduction --- p.102 / Chapter 3.1.2. --- Results --- p.104 / Chapter 3.1.3. --- Purification --- p.107 / Chapter 3.1.3.1. --- Cation-Exchange Chromatography on CM-Cellulose --- p.108 / Chapter 3.1.3.2. --- Affinity Chromatography on Affi-gel Blue gel --- p.111 / Chapter 3.1.3.3. --- Cation-Exchange Chromatography on Mono S Column --- p.114 / Chapter 3.1.3.4. --- Gel Filtration on Superdex 75 Column --- p.115 / Chapter 3.1.4. --- Characterization of Chitinase --- p.117 / Chapter 3.1.4.1. --- N-terminal Amino Acid Sequence --- p.117 / Chapter 3.1.4.2. --- Assay for Antifungal Activity --- p.118 / Chapter 3.1.4.3. --- Assay for Cell-Free Translation-inhibitory Activity --- p.120 / Chapter 3.1.4.4. --- Assay for HIV-1 Reverse Transcriptase Inhibitory Activity --- p.120 / Chapter 3.1.5. --- Characterization of Ribonuclease --- p.121 / Chapter 3.1.5.1. --- N-terminal Amino Acid Sequence --- p.121 / Chapter 3.1.5.2. --- Assay for Ribonuclease Activity --- p.122 / Chapter 3.1.5.3. --- Assay for Cell-Free Translation-inhibitory Activity --- p.125 / Chapter 3.1.5.4. --- Assay for Antifungal Activity --- p.125 / Chapter 3.1.5.5. --- Assay for Antiproliferative Activity --- p.126 / Chapter 3.1.6. --- Discussion --- p.127 / Chapter 3.2. --- Purification and Characterization of Ribosome-Inactivating Protein and Antifungal Protein from the mushroom Lyophyllum shimeji --- p.131 / Chapter 3.2.1. --- Introduction --- p.131 / Chapter 3.2.2. --- Results --- p.132 / Chapter 3.2.3. --- Purification --- p.134 / Chapter 3.2.3.1. --- Cation-Exchange Chromatography on CM-Cellulose --- p.135 / Chapter 3.2.3.2. --- Affinity Chromatography on Affi-gel Blue Gel --- p.137 / Chapter 3.2.3.3. --- Cation-Exchange Chromatography on Mono S --- p.140 / Chapter 3.2.4. --- Characterization of Ribosome-Inactivating Protein and Antifungal Protein from Lyophyllum shimeji --- p.142 / Chapter 3.2.4.1. --- N-terminal Amino Acid Sequence --- p.142 / Chapter 3.2.4.2. --- Assay for Antifungal Activity --- p.144 / Chapter 3.2.4.3. --- Assay for N-glycosidase Activity --- p.147 / Chapter 3.2.4.4. --- Assay for Mitogenic Activity --- p.147 / Chapter 3.2.4.5. --- Assay for HIV-1 Reverse Transcriptase Inhibitory Activity --- p.148 / Chapter 3.2.5. --- Discussion --- p.150 / Chapter 3.3. --- Purification and Characterization of Ribosome-inactivating Protein from the Hypsizigus marmoreus --- p.153 / Chapter 3.3.1. --- Introduction --- p.153 / Chapter 3.3.2. --- Result --- p.154 / Chapter 3.3.3. --- Purification --- p.155 / Chapter 3.3.3.1. --- Cation-Exchange Chromatography on CM-Cellulose --- p.156 / Chapter 3.3.3.2. --- Affinity-Chromatography on Affi-gel Blue Gel --- p.158 / Chapter 3.3.3.3. --- Anion-Exchange Chromatography on Mono Q Column --- p.160 / Chapter 3.3.4. --- Characterization of Ribosome-inactivating Protein from Hypsizigus marmoreus --- p.162 / Chapter 3.3.4.1. --- N-terminal Amino Acid Sequence --- p.162 / Chapter 3.3.4.2. --- Assay for Cell-Free Translation-Inhibiting Activity --- p.163 / Chapter 3.3.4.3. --- Assay for Antifungal Activity --- p.164 / Chapter 3.3.4.4. --- Assay for N-glycosidase Activity --- p.166 / Chapter 3.3.4.5. --- Assay for HIV-1 Reverse Transcriptase Inhibitory Activity --- p.166 / Chapter 3.3.4.6. --- Assay for mitogenic Activity --- p.167 / Chapter 3.3.4.7. --- Assay for Antiproliferative Activity --- p.167 / Chapter 3.3.5. --- Discussion --- p.159 / Chapter Chapter 4 --- General Discussion --- p.170 / References --- p.172

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323548
Date January 2001
ContributorsLam, Sze Kwan., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xxiv, 200 leaves : ill. ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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