Sub-2 Angstrom resolution structure determination using single-particle cryo-EM at 200 keV

IF 3.5 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Mengyu Wu , Gabriel C. Lander , Mark A. Herzik Jr.
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引用次数: 35

Abstract

Although the advent of direct electron detectors (DEDs) and software developments have enabled the routine use of single-particle cryogenic electron microscopy (cryo-EM) for structure determination of well-behaved specimens to high-resolution, there nonetheless remains a discrepancy between the resolutions attained for biological specimens and the information limits of modern transmission electron microscopes (TEMs). Instruments operating at 300 kV equipped with DEDs are the current paradigm for high-resolution single-particle cryo-EM, while 200 kV TEMs remain comparatively underutilized for purposes beyond sample screening. Here, we expand upon our prior work and demonstrate that one such 200 kV microscope, the Talos Arctica, equipped with a K2 DED is capable of determining structures of macromolecules to as high as ∼1.7 Å resolution. At this resolution, ordered water molecules are readily assigned and holes in aromatic residues can be clearly distinguished in the reconstructions. This work emphasizes the utility of 200 kV electrons for high-resolution single-particle cryo-EM and applications such as structure-based drug design.

Abstract Image

在200 keV下使用单粒子低温电镜进行亚2埃分辨率结构测定
尽管直接电子探测器(ded)的出现和软件的发展使得单粒子低温电子显微镜(cryo-EM)的常规使用能够对表现良好的样品进行高分辨率的结构测定,但生物样品的分辨率与现代透射电子显微镜(tem)的信息限制之间仍然存在差异。300 千伏的低温电子显微镜装备的仪器是目前高分辨率单粒子低温电子显微镜的范例,而200 千伏的低温电子显微镜在样品筛选以外的用途上仍然相对未得到充分利用。在这里,我们扩展了我们之前的工作,并证明了一个这样的200 千伏显微镜,配备K2 DED的Talos Arctica能够以高达1.7 Å的分辨率确定大分子的结构。在这个分辨率下,有序的水分子很容易被分配,并且在重建中可以清楚地区分芳香残基中的孔。这项工作强调了200 千伏电子在高分辨率单粒子低温电镜和基于结构的药物设计等应用中的效用。
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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