Crystallization and preliminary X-ray characterization of the Skp1-Fbg3 complex
Crystallization and preliminary X-ray characterization of the Skp1-Fbg3 complex
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Skp1-Fbg3 复合物的结晶和初步 X 射线表征
DOI：
10.1107/s1744309109050581
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
7番目)
中科院分区：
文献类型：
--
作者：
Kumanomidou;T.;et al.(員数9;7番目)
F-box 蛋白是 Skp1–Cullin1–F-box 蛋白–Rbx1 (SCF) 泛素连接酶复合物的底物识别成分。 Fbs1 是一种 F-box 蛋白，可与高甘露糖寡糖修饰的蛋白质特异性结合。 Fbg3 是另一种 F-box 蛋白，与 Fbs1 具有 51% 的序列同一性。尽管与寡糖结合所需的残基在 Fbs1 和 Fbg3 之间是保守的，但 Fbg3 不结合糖蛋白。 Skp1和Fbg3在大肠杆菌中共表达，其复合物被纯化至均质并结晶。微晶种与夹层悬滴技术相结合，提高了所得晶体的质量。该板状晶体属于P212121空间群，晶胞参数a = 34.1，b = 76.6，c = 193.9 Å，每个不对称单元有一个分子。
F-box proteins are the substrate-recognition components of Skp1–Cullin1–F-box protein–Rbx1 (SCF) ubiquitin ligase complexes. Fbs1, an F-box protein, binds specifically to proteins modified with high-mannose oligosaccharides. Fbg3, another F-box protein, has 51% sequence identity to Fbs1. Although the residues that are necessary for binding to oligosaccharides are conserved between Fbs1 and Fbg3, Fbg3 does not bind glycoproteins. Skp1 and Fbg3 were co-expressed in Escherichia coli and their complex was purified to homogeneity and crystallized. Microseeding combined with the sandwiched hanging-drop technique improved the quality of the resulting crystals. The plate-shaped crystals belonged to space group P212121, with unit-cell parameters a = 34.1, b = 76.6, c = 193.9 Å and one molecule per asymmetric unit.