| Summary: | 碩士 === 國立清華大學 === 生物科技研究所 === 94 === H. Pylori (Helicobacter pylori) is a Gram-negative, microaerophilic, and slow-growing bacterium, which can cause serious gastric ulcer, duodenum-associated disease and gastric cancer. According to the sequence alignment, HP0404 which is produced by H. pylori is the PKC inhibitor of HIT (histidine triad) protein family, because of the special sequence motif, His-X-His-X-His-X-X (X is a hydrophobic amino acid). HIT proteins fall into three branches, the Fhit (fragile histidine triad) branch that is found only in animals and fungi and histidine triad nucleotide-binding protein (Hint) branch that has represented in all cellular life and the Galt(Galactose-1-phosphate uridylytransferase) brench. Here we chose HP0404 as interest target to investigate the structure by using NMR spectroscopy (nuclear magnetic resonance). The C13- and N15-label protein sample of HP0404 have been produced for the NMR experiment. Proton, nitrogen, and carbon chemical shift assignment have been made for HP0404. assignment were made from a combination of homonuclear two-dimensional and N15- and C13- edited three –dimensional spectra at PH6.5 and 298K. Here we present the sequential assignment of HP0404 and make the residue annotation on the HSQC (heteronuclear single quantum correlation) spectra. We also did the ATP titration experiment for functional study and found residue I17, N20, S25, S44, K68, T81, N86 and H97 maybe important for interacting with ATP. This annotation maybe could offer slight help for further investigation between structure and function of HP0404.
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