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Purification and characterization of a 19 kDa zinc-binding protein in porcine brain.

by Wong Ping Shing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 97-112). / ACKNOWLEDGMENTS --- p.i / ABSTRACT --- p.ii / ABBREVIATIONS --- p.viii / Chapter 1. --- INTRODUCTION --- p.1 / Chapter 1.1 --- General properties of zinc / Chapter 1.1.1 --- Biochemistry of zinc --- p.2 / Chapter 1.1.2 --- Distribution of zinc in body --- p.3 / Chapter 1.1.3 --- Roles of zinc in protein function --- p.4 / Chapter 1.2 --- Zinc and zinc-binding proteins in brain / Chapter 1.2.1 --- Distribution of zinc in brain --- p.7 / Chapter 1.2.2 --- Metabolism of zinc in brain --- p.9 / Chapter 1.2.3 --- Compartments of zinc in brain --- p.10 / Chapter 1.2.4 --- Zinc-binding proteins in brain --- p.12 / Chapter 1.3 --- Pathological conditions of brain in relation to zinc --- p.15 / Chapter 1.4 --- Aim of the project --- p.20 / Chapter 2. --- MATERIALS AND METHODS --- p.22 / Chapter 2.1 --- Detection of zinc-binding proteins / Chapter 2.1.1 --- Sodium-Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.22 / Chapter 2.1.2 --- Electroblotting --- p.24 / Chapter 2.1.3 --- Radioactive zinc blotting --- p.25 / Chapter 2.1.4 --- Autoradiography --- p.25 / Chapter 2.2 --- Subcellular fractionation of porcine brain --- p.26 / Chapter 2.3 --- Purification and structural characterization of a 19 kDa zinc-binding protein / Chapter 2.3.1 --- Purification of a 19 kDa protein --- p.27 / Chapter 2.3.2 --- Sequencing of N-terminal blocked 19 kDa protein --- p.30 / Chapter 2.4 --- Characterization of the binding and biological properties of the 19 kDa zinc-binding protein / Chapter 2.4.1 --- Effect of divalent metal ions on zinc binding to the 19 kDa protein --- p.33 / Chapter 2.4.2 --- Effect of pH on the dissociation of radioactive zinc from the19 kDa protein --- p.34 / Chapter 2.4.3 --- Radioactive calcium blotting --- p.34 / Chapter 2.4.4 --- Interaction of radioactive zinc and radioactive calcium binding to the 19 kDa protein --- p.35 / Chapter 2.4.5 --- Calmodulin activity assay --- p.35 / Chapter 3. --- RESULTS / Chapter 3.1 --- Specificity of radioactive zinc-blot on zinc-binding protein detection --- p.38 / Chapter 3.2 --- Zinc-binding proteins in porcine brain --- p.38 / Chapter 3.3 --- Purification and identification of a cytosolic 19 kDa zinc- binding protein in porcine brain / Chapter 3.3.1 --- Zinc-dependent hydrophobic interaction chromatography --- p.44 / Chapter 3.3.2 --- N-terminal amino acid sequencing --- p.51 / Chapter 3.3.3 --- High pH native gel electrophoresis of 19 kDa protein --- p.51 / Chapter 3.4 --- The zinc and calcium binding properties of the 19 kDa protein / Chapter 3.4.1 --- Effect of pre-exposure to divalent cations on zinc binding --- p.54 / Chapter 3.4.2 --- Competition by divalent cations for zinc binding --- p.56 / Chapter 3.4.3 --- pH dependency of zinc dissociation --- p.56 / Chapter 3.4.4 --- Effect of zinc on radioactive calcium binding --- p.61 / Chapter 3.5 --- The biological activity of the 19 kDa protein / Chapter 3.5.1 --- Effect of the 19 kDa protein on the activity of calmodulin- dependent phosphodiesterase --- p.66 / Chapter 3.5.2 --- Effect of zinc on calmodulin-dependent phosphodiesterase activity --- p.69 / Chapter 3.5.4 --- "Effect of zinc on calcium-deficient, calmodulin-dependent phosphodiesterase activity" --- p.72 / Chapter 4. --- DISCUSSION / Chapter 4.1 --- Detection and Purification of zinc-binding proteins / Chapter 4.1.1 --- Strategy for the detection of zinc-binding proteins --- p.77 / Chapter 4.1.2 --- Purification of zinc-binding protein --- p.79 / Chapter 4.2 --- Amino acid sequencing of the 19 kDa protein --- p.82 / Chapter 4.3 --- Binding properties of the 19 kDa zinc-binding protein --- p.86 / Chapter 4.4 --- Effect of zinc and 19 kDa zinc-binding protein on calmodulin dependent phosphodiesterase --- p.92 / Chapter 4.5 --- Effect of zinc on the properties of calmodulin --- p.90 / Chapter 4.6 --- Significance of the ability of zinc to affect calmodulin activity --- p.94 / Chapter 5. --- CONCLUSION --- p.95 / Chapter 6. --- REFERENCES --- p.97

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_320537
Date January 1995
ContributorsWong, Ping Shing., 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, ix, 112 leaves : ill. (some col.) ; 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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