The amino-terminal structure of human fragile X mental retardation protein obtained using precipitant-immobilized imprinted polymers
The amino-terminal structure of human fragile X mental retardation protein obtained using precipitant-immobilized imprinted polymers
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DOI：
10.1038/ncomms7634
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发表时间：
2015-03-01
影响因子：
16.6
通讯作者：
Ren, Xueqin
中科院分区：
文献类型：
--
作者：
Hu, Yufeng;Chen, Zhenhang;Ren, Xueqin
柔性是蛋白质的内在属性，对于它们的生物学功能是必不可少的。然而，由于结构的灵活性，获得具有异质构象的蛋白质的高质量晶体仍然具有挑战性。在这里，我们展示了一种新的方法，将传统的沉淀剂吸附到分子印迹聚合物（MIPs）上，以促进蛋白质结晶，特别是对柔性蛋白质。通过应用该方法，获得了人脆性X智力低下蛋白的柔性N-末端的高质量晶体，其缺失导致最常见的遗传性智力低下。发现了一个新的KH结构域和一个分子间二硫键，并在溶液中发现了几种类型的二聚体，从而为该蛋白的功能提供了见解。此外，沉淀剂固定的分子印迹聚合物（piMIPs）成功地促进灵活的蛋白质晶体形成的五个模型蛋白质与增加的衍射分辨率。这突出了piMIP用于柔性蛋白质结晶的潜力。
Flexibility is an intrinsic property of proteins and essential for their biological functions. However, because of structural flexibility, obtaining high-quality crystals of proteins with heterogeneous conformations remain challenging. Here, we show a novel approach to immobilize traditional precipitants onto molecularly imprinted polymers (MIPs) to facilitate protein crystallization, especially for flexible proteins. By applying this method, high-quality crystals of the flexible N-terminus of human fragile X mental retardation protein are obtained, whose absence causes the most common inherited mental retardation. A novel KH domain and an intermolecular disulfide bond are discovered, and several types of dimers are found in solution, thus providing insights into the function of this protein. Furthermore, the precipitant-immobilized MIPs (piMIPs) successfully facilitate flexible protein crystal formation for five model proteins with increased diffraction resolution. This highlights the potential of piMIPs for the crystallization of flexible proteins.