Chan, King Lun Michel. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 111-121). / Abstracts in English and Chinese. / THESIS ASSESSMENT COMMITTEE --- p.i / ACKNOWLEDGEMENTS --- p.ii / 摘要 --- p.iv / ABSTRACT --- p.v / LIST OF ABBREVIATIONS --- p.xi / Chapter CHAPTER 1 --- INTRODUCTION / Chapter 1.1 --- Aldehyde Dehydrogenase Superfamily / Chapter 1.1.1 --- Classification and Substrate Specificities of Aldehyde Dehydrogenases --- p.1 / Chapter 1.1.2 --- Multiple Functions of Aldehyde Dehydrogenases and their Roles in Metabolism --- p.4 / Chapter 1.1.3 --- Structural Organization of Aldehyde Dehydrogenases in view of their Catalytic Mechanism --- p.7 / Chapter 1.2 --- Antiquitin / Chapter 1.2.1 --- Discovery and Plant Antiquitins --- p.15 / Chapter 1.2.2 --- Animal Antiquitins --- p.17 / Chapter 1.2.3 --- Human Antiquitin Gene Mutations and Pyridoxine-dependent Seizures --- p.19 / Chapter 1.2.4 --- Previous Findings on Seabream Antiquitin --- p.20 / Chapter 1.3 --- Aims of Study --- p.22 / Chapter CHAPTER 2 --- MATERIALS AND METHODS / Chapter 2.1 --- Materials / Chapter 2.1.1 --- "Subcloning, Expression and Purification of Human Antiquitin" --- p.24 / Chapter 2.1.2 --- Characterization of Human Antiquitin --- p.25 / Chapter 2.1.3 --- "Crystallization of Human Antiquitin, Diffraction Data Collection and Structure Determination" --- p.26 / Chapter 2.1.4 --- Mutational Studies of Human Antiquitin --- p.26 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- "Subcloning, Expression and Purification of Human Antiquitin" / Chapter 2.2.1.1 --- Subcloning of the Full-length Human Antiquitin cDNA --- p.27 / Chapter 2.2.1.2 --- Bacterial Expression of Recombinant Human Antiquitin --- p.29 / Chapter 2.2.1.3 --- Purification of Human Antiquitin --- p.30 / Chapter 2.2.2 --- Characterization of Human Antiquitin / Chapter 2.2.2.1 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.31 / Chapter 2.2.2.2 --- Size-exclusion Chromatography - Multi-angle Light Scattering --- p.32 / Chapter 2.2.2.3 --- Isoelectric Focusing --- p.33 / Chapter 2.2.2.4 --- pH-rate Profile --- p.33 / Chapter 2.2.2.5 --- Stability Studies --- p.34 / Chapter 2.2.2.6 --- Substrate Specificity Study --- p.35 / Chapter 2.2.3 --- "Crystallization of Human Antiquitin, Diffraction Data Collection and Structure Determination" / Chapter 2.2.3.1 --- Crystallization of Human Antiquitin --- p.36 / Chapter 2.2.3.2 --- Diffraction Data Collection and Model Building --- p.37 / Chapter 2.2.4 --- Mutational Studies of Human Antiquitin / Chapter 2.2.4.1 --- Preparation of Mutant Plasmids --- p.39 / Chapter 2.2.4.2 --- "Expression, Purification and Kinetics Studies of Mutants" --- p.39 / Chapter CHAPTER 3 --- RESULTS / Chapter 3.1 --- "Subcloning, Expression and Purification of Human Antiquitin" --- p.43 / Chapter 3.2 --- Characterization of Human Antiquitin --- p.49 / Chapter 3.3 --- "Crystallization of Human Antiquitin, Diffraction Data Collection and Structure Determination" / Chapter 3.3.1 --- "Crystallization, Data Collection and Refinement" --- p.59 / Chapter 3.3.2 --- Human Antiquitin Structure --- p.63 / Chapter 3.4 --- Mutational Studies of Human Antiquitin --- p.75 / Chapter CHAPTER 4 --- DISCUSSION / Chapter 4.1 --- Characterization and Substrate Specificity of Recombinant Human Antiquitin --- p.83 / Chapter 4.2 --- Crystallization and Crystal Structure of Human Antiquitin / Chapter 4.2.1 --- Crystallization --- p.86 / Chapter 4.2.2 --- Overall structure --- p.86 / Chapter 4.2.3 --- Cofactor binding --- p.88 / Chapter 4.2.4 --- Substrate Binding and Catalysis --- p.91 / Chapter 4.3 --- Mutations in Human Antiquitin Gene and their Relationship with Pyridoxine-dependent Seizures --- p.95 / Chapter 4.4 --- Comparison of Antiquitin Gene and Other Aldehyde Dehydrogenases --- p.104 / Chapter CHAPTER 5 --- FUTURE PROSPECTS --- p.106 / LIST OF REFERENCES --- p.111 / APPENDIX --- p.122
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326171 |
| Date | January 2007 |
| Contributors | Chan, King Lun Michel., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xii, 122 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use 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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