Crystallization and preliminary crystallographic anal-ysis of human LR11 Vps10p domain
Crystallization and preliminary crystallographic anal-ysis of human LR11 Vps10p domain
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人LR11 Vps10p结构域的结晶及初步晶体学分析
DOI：
10.1107/s1744309110048153
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Takagi J
中科院分区：
文献类型：
--
作者：
Nakata Z;Nagae M;Yasui N;Bujo H;Nogi T;Takagi J
与11个结合重复序列（LR 11;也称为sorLA）相关的低密度脂蛋白受体（LDLR）在遗传上与迟发性阿尔茨海默病相关，并且被认为参与神经退行性过程。LR 11含有一个液泡蛋白分选10蛋白（Vps 10 p）结构域。由于该结构域与其他受体中的蛋白质-蛋白质相互作用有关，因此其结构和功能具有重要的生物学意义。在哺乳动物细胞中表达人LR 11 Vps 10 p结构域，并使用悬滴气相扩散法结晶纯化的蛋白。样品的酶促去糖基化是获得衍射质量晶体的关键。去糖基化的LR 11 Vps 10 p-结构域晶体属于六方空间群P6122。收集衍射数据集至2.4 μ m分辨率，并获得澄清的分子置换溶液。
Low-density lipoprotein receptor (LDLR) relative with 11 binding repeats (LR11; also known as sorLA) is genetically associated with late-onset Alzheimer's disease and is thought to be involved in neurodegenerative processes. LR11 contains a vacuolar protein-sorting 10 protein (Vps10p) domain. As this domain has been implicated in protein–protein interaction in other receptors, its structure and function are of great biological interest. Human LR11 Vps10p domain was expressed in mammalian cells and the purified protein was crystallized using the hanging-drop vapour-diffusion method. Enzymatic deglycosylation of the sample was critical to obtaining diffraction-quality crystals. Deglycosylated LR11 Vps10p-domain crystals belonged to the hexagonal space group P6122. A diffraction data set was collected to 2.4 Å resolution and a clear molecular-replacement solution was obtained.