Characterization of the Binding of Spike H Protein of Bacteriophage .PHI.X174 with Receptor Lipopolysaccharides.
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概要
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The spike H protein of bacteriophage φX 174 was prepared as a hexa histidine-tagged fusion (HisH). On enzyme-linked plate assaying, HisH was found to bind specifically to the lipopolysaccharides (LPSs) of φX174-sensitive strains, Escherichia coli C and Salmonella typhimurium Ra chemotype, having the complete oligosaccharide sequence of the R-core on the LPSs. In sharp contrast, HisH bound weakly to the LPSs of ΦX174-insensitive strains, i.e. E. coli F 583 (Rd2) lacking some terminal saccharides and E. coli O111:B4 (smooth strain) having additional O-repeats on the R-core. The fluorescence spectra of HisH changed dose-dependently in the case of the LPS of E. coli C, the intensity increasing and the emission peak shifting to the shorter wavelength side, which was attributable to the hydrophobic interaction of HisH with the LPS. The binding equilibrium was analyzed by fluorometric titration to determine the dissociation constant Kd, 7.02±0.37 μM, and the Gibbs free energy change ΔG0, -29.1 kJ mol-1 (at 22°C, pH 7.4). Based on the temperature dependence of Kd in a van't Hoff plot, the standard enthalpy change ΔH0 and the entropy change. ΔS0 were calculated to be +23.7 kJ mol-1 and 179 J mol-1 K-1 at 22°C, respectively, and this binding was thereby concluded to be an entropy-driven reaction.
- 社団法人 日本生化学会の論文
著者
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Suzuki Ryoko
Department Of Biomolecular Science Faculty Of Science Toho University
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KARITA Shuichi
Center for Molecular Biology and Genetics Mie University
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Kawaura Tomoko
Department Of Life Science Faculty Of Bioresources Mie University
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Kashimura Naoki
Department Of Bioscience Mie University
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Inagaki Minoru
Department Of Life Science Faculty Of Bioresources Mie University
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Tanaka Akiyoshi
Department Of Chemistry Faculty Of Education Mie University
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Wakashima Hirohito
Department Of Bioscience Faculty Of Bioresources And Center For Molecular Biology And Genetics Mie U
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Suzuki Ryoko
Department of Bioscience, Faculty of Bioresources
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Nishikawa Shiro
Department of Bioscience, Faculty of Bioresources
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