Abstract
In recent years, the structural genomics has been studied and developed thoroughly. Genomic sequencing information has combined the latest molecular and structural biology; hence scientists are able to move one step further in the structural genomics. Helicobacter pylori is a Gram-negative, microaerophilic, and slow-growing bacterium with flagellum and spiral appearance. It causes serious gastric ulcer and duodenum-associated diseases, and almost half population of the world has been infected by this bacterium. Since scientists have little understanding in the proteins produced by Helicobacter pylori genomics so far, we have chosen this bacterium as the major subject of the structural genomic project. The aim of this thesis is to investigate a protein called HP0404 from Hlicobacter pylori genomics. The results from amino acid sequence alignment inferred that its biological activity might be inhibition of PKC. The secondary structure and stability of the protein were determined by circular dichroism, and it revealed that HP0404 is a heat stable protein. In addition, fluorescence spectrometer was used to detect the binding ability between the protein and metal ions, and the quantitative analysis of the metal ions within the protein was performed by ICP-MS, thus it is shown that HP0404 has specific binding with Zn2+. Furthermore, the non- radioactive method was applied to demonstrate that HP0404 could effectively suppress the PKC activity. The difference between the protein structures of HP0404 and HP0404 C14A was examined by NMR. Moreover, we attempted to use X-ray technology to study the protein structure of HP0404. The hanging-drop method was applied to crystallize HP0404, and the structure would be determined by X-ray. The purpose was to study the molecular structure of HP0404 in a different way.