by Tsang Yuen Shan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 98-110). / ACKNOWLEDGMENTS --- p.I / ABSTRACT --- p.II / CONTENTS --- p.IV / ABBREVIATIONS --- p.VII / LIST OF FIGURES --- p.IX / LIST OF TABLES --- p.XII / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 1.1 --- Biochemistry of zinc --- p.1 / Chapter 1.2 --- Zinc within the body --- p.2 / Chapter 1.2.1 --- Roles of zinc in general biochemical processes --- p.4 / Chapter 1.2.2 --- Zinc and zinc-binding proteins (ZnBPs) --- p.9 / Chapter 1.3 --- Zinc in brain --- p.12 / Chapter 1.3.1 --- Distribution of zinc in brain --- p.13 / Chapter 1.3.2 --- Roles of zinc in brain 、 --- p.14 / Chapter 1.4 --- Summary --- p.18 / Chapter 1.5 --- Aim of Study --- p.20 / Chapter 2. --- MATERIALS AND METHODS --- p.22 / Chapter 2.1 --- Detection of zinc binding proteins --- p.22 / Chapter 2.1.1 --- Experimental animal --- p.22 / Chapter 2.1.2 --- Subcellular fractionation of porcine brain --- p.22 / Chapter 2.1.2 --- Determination of protein concentration --- p.24 / Chapter 2.1.3 --- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) --- p.24 / Chapter 2.1.4 --- Two dimensional polyacrylmide gel electrophoresis (2D-PAGE) --- p.25 / Chapter 2.1.5 --- "Fixing, staining and destaining" --- p.28 / Chapter 2.1.6 --- Western blotting for SDS-PAGE --- p.29 / Chapter 2.1.7 --- 65Zn binding --- p.30 / Chapter 2.1.8 --- Autoradiography --- p.30 / Chapter 2.1.9 --- Data quantification --- p.31 / Chapter 2.2 --- Identification of the 21 kD ZnBP --- p.31 / Chapter 2.2.1 --- N-terminal sequencing of the 21 kD ZnBP --- p.31 / Chapter 2.2.2 --- In situ cyanogen bromide (CNBr) cleavage --- p.32 / Chapter 2.2.3 --- Tricine SDS-PAGE --- p.33 / Chapter 2.2.4 --- Isolation of myelin --- p.34 / Chapter 2.2.5 --- Extraction of the 21 kD ZnBP --- p.35 / Chapter 2.2.6 --- Comparison of the mobility of the 21 kD ZnBP with that of bovine myelin basic protein (MBP) in three different gel electrophoresis system --- p.35 / Chapter 2.3 --- Characterization of the zinc binding properties of MBP --- p.38 / Chapter 2.3.1 --- Concentration-dependence of MBP on 65Zn binding --- p.38 / Chapter 2.3.2 --- Effect of other divalent cations on 65Zn binding to the 21 kD ZnBP --- p.38 / Chapter 2.3.3 --- Effect of pH on 65Zn binding to the 21 kD ZnBP --- p.39 / Chapter 2.3.4 --- Effect of modification of histidine residues in MBP on 65Zn binding --- p.39 / Chapter 2.4 --- Effect of zinc on the interaction between MBP and membrane- lipid --- p.40 / Chapter 2.4.1 --- Effect of zinc on MBP-induced phospholipid vesicle aggregation --- p.40 / Chapter 2.4.2 --- Effect of some divalent cations on MBP-induced phospholipid vesicle aggregation --- p.41 / Chapter 3. --- RESULTS --- p.42 / Chapter 3.1 --- Distribution of ZnBPs in porcine brain --- p.42 / Chapter 3.1.1 --- Subcellular distribution of ZnBPs in porcine brain --- p.42 / Chapter 3.1.2 --- "Comparison of the subcellular distribution of ZnBPs in brain, heart and liver" --- p.45 / Chapter 3.1.3 --- Regional distribution of ZnBPs in porcine brain --- p.48 / Chapter 3.1.4 --- Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of total brain homogenates --- p.51 / Chapter 3.2 --- Identification of the 21 kD ZnBP --- p.58 / Chapter 3.2.1 --- N-terminal sequencing of the 21 kD ZnBP --- p.58 / Chapter 3.2.2 --- CNBr cleavage of the 21 kD ZnBP and the N-terminal sequencing of CNBr cleaved fragment --- p.59 / Chapter 3.2.3 --- Detection of the 21 kD ZnBP in myelin fraction and extraction of the 21 kD ZnBP by chloroform-methanol extraction --- p.59 / Chapter 3.2.4 --- Comparison of properties of the 21 kD ZnBP with bovine MBP on three electrophoresis systems --- p.61 / Chapter 3.2.5 --- CNBr cleavage patterns of the 21 kD ZnBP and the bovine MBP --- p.64 / Chapter 3.3 --- Characterization of the zinc binding properties of MBP --- p.69 / Chapter 3.3.1 --- Concentration-dependence of MBP on 65Zn binding --- p.69 / Chapter 3.3.2 --- Effect of other divalent cations on 65Zn binding to the 21 kD ZnBP --- p.69 / Chapter 3.3.3 --- Effect of pH on 65Zn binding to the 21 kD ZnBP --- p.71 / Chapter 3.3.4 --- Effect of modification of histidine residues in MBP on 65Zn binding --- p.73 / Chapter 3.4 --- Effect of zinc on the interaction between MBP and membrane- lipid --- p.76 / Chapter 3.4.1 --- Effect of zinc on MBP-induced phospholipid vesicle aggregation --- p.76 / Chapter 3.4.2 --- Effect of other divalent cations on MBP-induced phospholipid vesicle aggregation --- p.81 / Chapter 4. --- DISCUSSION --- p.83 / Chapter 5. --- CONCLUSIONS --- p.96 / Chapter 6. --- REFERENCES --- p.98
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_320669 |
| Date | January 1994 |
| Contributors | Tsang, Yuen Shan., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Publisher | Chinese University of Hong Kong |
| Source Sets | The Chinese University of Hong Kong |
| Language | English |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xii, 110 leaves : ill. (some mounted) ; 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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