A study of the ultrastructure of fragile-X-related proteins.
A study of the ultrastructure of fragile-X-related proteins.
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DOI：
10.1042/bj20082197
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发表时间：
2009-04-15
期刊：
影响因子：
--
通讯作者：
Pastore A
中科院分区：
文献类型：
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作者：
Sjekloća L;Konarev PV;Eccleston J;Taylor IA;Svergun DI;Pastore A
脆性X相关蛋白形成一个涉及RNA代谢的家族。它们的序列由保守的N-末端和含有Tudor和KH结构域的中心区域以及具有富含精氨酸和甘氨酸残基的基序的不同C-末端组成。该家族中研究最广泛的成员可能是FMRP（脆性X智力低下蛋白），因为人类缺乏或突变这种蛋白会导致脆性X综合征，这是遗传性智力低下的最常见原因。了解FMRP的结构特性对于将其与其功能相关联至关重要。FMRP的分离结构域的结构已被报道，但关于不同模块的空间排列尚不清楚，部分原因是难以大量生产全长蛋白质及其多结构域片段，纯度和单分散性适合于结构研究。在本研究中，我们描述了我们如何产生重叠的重组片段的人FMRP及其旁系同源物，其中包括进化保守区。我们已经研究了他们的行为在解决方案中的补充生物化学和生物物理技术，确定了区域，促进自我协会，并确定其整体的三维形状。本研究为进一步的研究铺平了道路，并合理化现有的知识，这些蛋白质的自缔合特性。
Fragile-X-related proteins form a family implicated in RNA metabolism. Their sequence is composed of conserved N-terminal and central regions which contain Tudor and KH domains and of a divergent C-terminus with motifs rich in arginine and glycine residues. The most widely studied member of the family is probably FMRP (fragile X mental retardation protein), since absence or mutation of this protein in humans causes fragile X syndrome, the most common cause of inherited mental retardation. Understanding the structural properties of FMRP is essential for correlating it with its functions. The structures of isolated domains of FMRP have been reported, but nothing is yet known with regard to the spatial arrangement of the different modules, partly because of difficulties in producing both the full-length protein and its multidomain fragments in quantities, purities and monodispersity amenable for structural studies. In the present study, we describe how we have produced overlapping recombinant fragments of human FMRP and its paralogues which encompass the evolutionary conserved region. We have studied their behaviour in solution by complementary biochemical and biophysical techniques, identified the regions which promote self-association and determined their overall three-dimensional shape. The present study paves the way to further studies and rationalizes the existing knowledge on the self-association properties of these proteins.