I:人類 6-磷酸葡萄糖異構酵素與鳥苷三磷酸結合及其遺傳性突變之結構與功能關聯性研究II: 竹嵌紋病毒戴帽酵素中保留性組胺酸於形成[酵素-m7GMP]中間產物時之功能分析

Abstract

The part I in this dissertation is “Structure-function relationship of human phosphoglucose isomerase − A study bases on inherited mutations and GTP-binding”. This part contains two chapters. Chapter I entitled “Effects of inherited mutations on catalytic activity and structural stability of human phosphoglucose isomerase expressed in Escherichia coli” demonstrates the effects of the inherited mutations associated with nonspherocytic hemolytic anemia on the protein properties including kinetic parameter and protein stability. Chapter II entitled “Characterization of the novel binding of GTP to human phosphoglucose isomerase/autocrine motility factor” explicates a novel binding between human phosphoglucose isomerase and GTP. The biochemical and biological characterization of this binding is described in this chapter. The part II is “Functional analysis of the conserved histidine residue of Bamboo mosaic virus capping enzyme in the activity for the covalent [Enzyme-m7GMP] intermediate formation”. In this part, the amino acid residue that covalently bond to m7GMP was identified using hydroxylamine digestion mapping and mutagenesis assay. The roles of the histidine residue, H66, and the conserved histidine residue, H68, on the formation of [Enzyme-m7GMP] intermediate were also investigated.