Reconstruction of Average Subtracted Tubular Regions (RASTR) enables structure determination of tubular filaments by cryo-EM

IF 3.5 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Peter S. Randolph , Scott M. Stagg
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引用次数: 1

Abstract

As the field of electron microscopy advances, the increasing complexity of samples being produced demand more involved processing methods. In this study, we have developed a new processing method for generating 3D reconstructions of tubular structures. Tubular biomolecules are common throughout many cellular processes and are appealing targets for biophysical research. Processing of tubules with helical symmetry is relatively straightforward for electron microscopy if the helical parameters are known, but tubular structures that deviate from helical symmetry (asymmetrical components, local but no global order, etc) present myriad issues. Here we present a new processing technique called Reconstruction of Average Subtracted Tubular Regions (RASTR), which was developed to reconstruct tubular structures without applying symmetry. We explain the RASTR approach and quantify its performance using three examples: a simulated symmetrical tubular filament, a symmetrical tubular filament from cryo-EM data, and a membrane tubule coated with locally ordered but not globally ordered proteins.

Abstract Image

平均减去管状区域(RASTR)的重建使管状细丝的结构通过低温电镜测定
随着电子显微镜领域的发展,所生产的样品越来越复杂,需要更复杂的处理方法。在这项研究中,我们开发了一种新的处理方法来生成管状结构的三维重建。管状生物分子在许多细胞过程中都很常见,是生物物理研究的重要目标。如果螺旋参数已知,用电子显微镜处理螺旋对称的小管是相对简单的,但是偏离螺旋对称的管状结构(不对称成分,局部但没有全局秩序等)存在无数问题。在这里,我们提出了一种新的处理技术,称为平均减去管状区域重建(RASTR),该技术是在不应用对称的情况下重建管状结构的。我们通过三个例子解释了RASTR方法并量化了其性能:模拟对称管状细丝,来自低温电镜数据的对称管状细丝,以及涂有局部有序而非全局有序蛋白质的膜小管。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Structural Biology: X
Journal of Structural Biology: X Biochemistry, Genetics and Molecular Biology-Structural Biology
CiteScore
6.50
自引率
0.00%
发文量
20
审稿时长
62 days
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