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Expression purification crystallization and preliminary crystallographic study of the cytoplasmic domain of the mitochondrial dynamics protein MiD51

机译:线粒体动力学蛋白MiD51的胞质域的表达纯化结晶和初步晶体学研究

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摘要

Mitochondria play central roles in many cellular and physiological processes. They are highly dynamic organelles and continually undergo fusion and fission. Mitochondrial dynamics protein 51 kDa (MiD51), an integral mitochondrial outer membrane protein, recruits dynamin-related protein 1 (Drp1; a mitochondrial fission protein) to mitochondria and facilitates Drp1-directed mitochondrial fission. In this study, the cytoplasmic domain of MiD51 was overexpressed in Escherichia coli, purified and crystallized. An X-ray diffraction data set was collected to a resolution of 3.1 Å and the crystal belonged to space group P41212, with unit-cell parameters a = b = 90.1, c = 124.7 Å, α = β = γ = 90°. The asymmetric unit had the highest probability of containing one molecule, with a Matthews coefficient of 3.32 Å3 Da−1 and a solvent content of 63.0%.
机译:线粒体在许多细胞和生理过程中起着核心作用。它们是高度动态的细胞器,并不断发生融合和裂变。线粒体动力学蛋白51 kDa(MiD51)是线粒体的外膜蛋白,可将动力蛋白相关蛋白1(Drp1;线粒体裂变蛋白)募集到线粒体,并促进Drp1定向的线粒体裂变。在这项研究中,MiD51的胞质域在大肠杆菌中过表达,纯化和结晶。收集到的X射线衍射数据集的分辨率为3.1,晶体属于空间群P41212,其晶胞参数a = b = 90.1,c = 124.7,α=β=γ= 90°。不对称单元包含一个分子的可能性最高,其马修斯系数为3.32Å 3 Da -1 ,溶剂含量为63.0%。

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