Characterization of the Binding of Spike H Protein of Bacteriophage .PHI.X174 with Receptor Lipopolysaccharides.
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- Inagaki Minoru
- Department of Bioscience, Faculty of Bioresources
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- Tanaka Akiyoshi
- Department of Bioscience, Faculty of Bioresources
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- Suzuki Ryoko
- Department of Bioscience, Faculty of Bioresources
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- Wakashima Hirohito
- Department of Bioscience, Faculty of Bioresources
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- Kawaura Tomoko
- Department of Bioscience, Faculty of Bioresources
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- Karita Shuichi
- Center for Molecular Biology and Genetics, Mie University
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- Nishikawa Shiro
- Department of Bioscience, Faculty of Bioresources
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- Kashimura Naoki
- Department of Bioscience, Faculty of Bioresources
抄録
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.
収録刊行物
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- The Journal of Biochemistry
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The Journal of Biochemistry 127 (4), 577-583, 2000
The Japanese Biochemical Society
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詳細情報 詳細情報について
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- CRID
- 1571698603079529216
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- NII論文ID
- 130003533839
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- ISSN
- 0021924X
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- 本文言語コード
- en
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- データソース種別
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- CiNii Articles