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Studies on zinc-binding proteins in porcine brain.

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

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_320669
Date January 1994
ContributorsTsang, Yuen Shan., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
PublisherChinese University of Hong Kong
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xii, 110 leaves : ill. (some mounted) ; 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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