Crystallization and preliminary X-ray crystallographic analysis of eIF5BΔN and the eIF5BΔN-eIF1AΔN complex
Crystallization and preliminary X-ray crystallographic analysis of eIF5BΔN and the eIF5BΔN-eIF1AΔN complex
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eIF5BΔN 和 eIF5BΔN-eIF1AΔN 复合物的结晶和初步 X 射线晶体学分析
DOI：
10.1107/s1744309111015910
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
Isao Tanaka and Min Yao
中科院分区：
文献类型：
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作者：
Aiping Zheng;Reo Yamamoto;Masaaki Sokabe;Isao Tanaka and Min Yao
两个普遍保守的翻译起始因子 eIF5B 和 eIF1A 之间的结合对于核糖体上真核蛋白质合成的起始步骤非常重要。通过这种相互作用，eIF1A 协助将 eIF5B 招募到起始 40S 亚基中；然后，eIF5B 会促进 60S 亚基的连接，形成起始 80S 核糖体。在此，报告了来自酿酒酵母的 eIF5BΔN 和 eIF5BΔN-eIF1AΔN 复合物的表达、纯化、结晶和初步 X 射线分析。 eIF5BΔN晶体的衍射分辨率为2.45 Å，属于P41212空间群，晶胞参数a = b = 130.0，c = 71.7 Å。据估计，该不对称单元包含一个分子。通过 Se-SAD 获得初始相。 eIF5BΔN–eIF1AΔN 复合物的晶体衍射分辨率为 3.3 Å，属于 P212121 空间群，晶胞参数 a = 101.9、b = 120.9、c = 132.8 Å。据估计，不对称单元包含两个复杂分子。
The binding between two universally conserved translation initiation factors, eIF5B and eIF1A, is important in the initiation step of eukaryotic protein synthesis on the ribosome. Through this interaction, eIF1A assists in recruiting eIF5B to the initiating 40S subunit; eIF5B then encourages the joining of the 60S subunit to form an initiating 80S ribosome. Here, the expression, purification, crystallization and preliminary X-ray analyses of eIF5BΔN and the eIF5BΔN–eIF1AΔN complex from Saccharomyces cerevisiae are reported. The crystal of eIF5BΔN diffracted to 2.45 Å resolution and belonged to space group P41212, with unit-cell parameters a = b = 130.0, c = 71.7 Å. The asymmetric unit was estimated to contain one molecule. The initial phase was obtained by Se-SAD. The crystal of the eIF5BΔN–eIF1AΔN complex diffracted to 3.3 Å resolution and belonged to space group P212121, with unit-cell parameters a = 101.9, b = 120.9, c = 132.8 Å. The asymmetric unit was estimated to contain two complex molecules.