Crystallization and Preliminary X-Ray Diffraction Study of Recombinant Human Eukaryotic Initiation Factor-4E.
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概要
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Recombinant human eukaryotic initiation factor-4E (eIF-4E), purified by m7GTP-Sepharose 4B affinity chromatography, was used for crystallization. After concentration of the eIF-4E protein (7mg/ml), the solution was subjected to crystallization by the hanging-drop method. Transparent needle crystals complexed with m7GTP were obtained from 50mM 2-(N-morpholino)ethanesulfonic acid-KOH buffer (pH 6.5) containing 25% (w/v) polyethylene glycol 6000 and 0.2M (NH4)2SO4. The crystals belong to tetragonal space group P41 or P43, of Z=4, with unit-cell dimensions of a=89.26, b=89.26, and c=38.51 Å, and diffract beyond 2.1 Å resolution. The Vm value was calculated to be 3.07 Å3/Da, which indicates a solvent content of 59.9%.
- 社団法人 日本生化学会の論文
著者
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Doi Mitsunobu
Department Of Physical Chemistry Osaka University Of Pharmaceutical Sciences
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KITAMURA Kunihiro
Research Center of Taisho Pharmaceutical Co., Ltd.
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Ishida Toshimasa
Department Of Physical Chemistry Osaka University Of Pharmaceutical Sciences
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Tomoo Koji
Department Of Physical Chemistry Osaka University Of Pharmaceutical Sciences
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Okabe Koumei
Department Of Pyysical Chemistry Osaka University Of Pharmaceutical Sciences
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Nishi Noriko
Department Of Biology Faculty Of Science Ochanomizu University
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Morino Shigenobu
Department Of Physical Chemistry Osaka University Of Pharmaceutical Sciences
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Morino Shigenobu
Department of Physical Chemistry, Osaka University of Pharmaceutical Sciences
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