Fragment-based phase extension for three-dimensional structure determination of membrane proteins by electron crystallography.

Fragment-based phase extension for three-dimensional structure determination of membrane proteins by electron crystallography.
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DOI:
10.1016/j.str.2011.04.008
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发表时间:
2011-07-13
期刊:
影响因子:
5.7
通讯作者:
Gonen, Tamir
Gonen, Tamir
中科院分区:
生物学2区
文献类型:
--
作者:
Wisedchaisri, Goragot;Gonen, Tamir

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在电子晶体学中,膜蛋白质结构是由蛋白质嵌入膜中的二维晶体确定的。一旦生长出大而有序的2D晶体,电子晶体学中的瓶颈之一是图像数据的收集,以直接提供高分辨率的实验阶段。在这里,我们描述了一种新的方法来绕过这个瓶颈,消除了对高分辨率成像的需要。我们利用电子晶体学的优势,从低分辨率图像中快速获得准确的实验相位信息,并从电子衍射中获得准确的高分辨率振幅信息。低分辨率实验相位用于放置α-螺旋片段,并使用片段相位扩展到高分辨率。通过对高分辨率衍射数据进行密度修改,然后进行片段扩展和结构细化,进一步改善了相位。使用这种方法,三种膜蛋白的结构被快速,准确地确定到原子分辨率没有高分辨率的图像数据。
In electron crystallography membrane protein structure is determined from two-dimensional crystals where the protein is embedded in a membrane. Once large and well-ordered 2D crystals are grown one of the bottlenecks in electron crystallography is the collection of image data to directly provide experimental phases to high resolution. Here we describe a new approach to bypass this bottleneck, eliminating the need for high-resolution imaging. We use the strengths of electron crystallography in rapidly obtaining accurate experimental phase information from low-resolution images and accurate high-resolution amplitude information from electron diffraction. The low-resolution experimental phases were used for the placement of α-helix fragments and extended to high resolution using phases from the fragments. Phases were further improved by density modifications followed by fragment expansion and structure refinement against the high-resolution diffraction data. Using this approach, structures of three membrane proteins were determined rapidly and accurately to atomic resolution without high-resolution image data.
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发表时间: 2004-05-13
期刊: NATURE
影响因子: 64.8
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