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首页> 外文期刊>Nucleic Acids Research >3.9 angstrom structure of the nucleosome core particle determined by phase-plate cryo-EM
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3.9 angstrom structure of the nucleosome core particle determined by phase-plate cryo-EM

机译：3.9埃的相板冷冻-电磁法测定核小体核心颗粒的结构

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The Volta phase plate is a recently developed electron cryo-microscopy (cryo-EM) device that enables contrast enhancement of biological samples. Here we have evaluated the potential of combining phase-plate imaging and single particle analysis to determine the structure of a small protein-DNA complex. To test the method, we made use of a 200 kDa Nucleosome Core Particle (NCP) reconstituted with 601 DNA for which a high-resolution X-ray crystal structure is known. We find that the phase plate provides a significant contrast enhancement that permits individual NCPs and DNA to be clearly identified in amorphous ice. The refined structure from 26,060 particles has an overall resolution of 3.9 angstrom and the density map exhibits structural features consistent with the estimated resolution, including clear density for amino acid side chains and DNA features such as the phosphate backbone. Our results demonstrate that phase-plate cryo-EM promises to become an important method to determine novel near-atomic resolution structures of small and challenging samples, such as nucleosomes in complex with nucleosome-binding factors.
机译：Volta相板是最近开发的电子冷冻显微镜（cryo-EM）设备，可增强生物样品的对比度。在这里，我们评估了相板成像和单颗粒分析相结合以确定小蛋白质-DNA复合物结构的潜力。为了测试该方法，我们使用了以601 DNA重组的200 kDa核小体核心颗粒（NCP），已知该DNA具有高分辨率的X射线晶体结构。我们发现，相板提供了显着的对比度增强，使单个NCP和DNA可以在无定形冰中清晰识别。从26,060个颗粒中提炼的结构的总分辨率为3.9埃，密度图显示的结构特征与估算的分辨率一致，包括氨基酸侧链的清晰密度和DNA特征（例如磷酸骨架）。我们的结果表明，相板式冷冻EM有望成为确定小而富挑战性样品（例如与核小体结合因子复合的核小体）的新型近原子拆分结构的重要方法。

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