钾离子通道的大尺寸和高度有序二维晶体的生长，脂环境的结构作用。
Growth of large and highly ordered 2D crystals of a K⁺ channel, structural role of lipidic environment.
机构信息
Laboratory of Molecular Biophysics, Department of Biochemistry, University of Oxford, Oxford, United Kingdom.
出版信息
Biophys J. 2013 Jul 16;105(2):398-408. doi: 10.1016/j.bpj.2013.05.054.
Abstract
2D crystallography has proven to be an excellent technique to determine the 3D structure of membrane proteins. Compared to 3D crystallography, it has the advantage of visualizing the protein in an environment closer to the native one. However, producing good 2D crystals is still a challenge and little statistical knowledge can be gained from literature. Here, we present a thorough screening of 2D crystallization conditions for a prokaryotic inwardly rectifying potassium channel (>130 different conditions). Key parameters leading to very large and well-organized 2D crystals are discussed. In addition, the problem of formation of multilayers during the growth of 2D crystals is also addressed. An intermediate resolution projection map of KirBac3.1 at 6 Å is presented, which sheds (to our knowledge) new light on the structure of this channel in a lipid environment.
摘要
2D 晶体学已被证明是一种确定膜蛋白 3D 结构的出色技术。与 3D 晶体学相比，它具有在更接近天然环境中可视化蛋白质的优势。然而，产生良好的 2D 晶体仍然是一个挑战，并且从文献中几乎无法获得统计学知识。在这里，我们对一种原核内向整流钾通道（>130 种不同条件）的 2D 结晶条件进行了全面筛选。讨论了导致非常大且组织良好的 2D 晶体的关键参数。此外，还解决了在 2D 晶体生长过程中形成多层的问题。呈现了 KirBac3.1 在 6 Å 分辨率下的中间投影图，这为该通道在脂质环境中的结构提供了（据我们所知）新的认识。
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