Mapping Single-Molecule Protein Complexes in 3D with DNA Nanoswitch Calipers.

Mapping Single-Molecule Protein Complexes in 3D with DNA Nanoswitch Calipers.
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用DNA纳米开关卡尺在3D中绘制单分子蛋白质复合物。

DOI:
10.1021/jacs.3c10262
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发表时间:
2023-12-27
影响因子:
15
通讯作者:
Wong, Wesley P.
Wong, Wesley P.
中科院分区:
化学1区
文献类型:
--
作者:
Shrestha, Prakash;Yang, Darren;Ward, Andrew;Shih, William M.;Wong, Wesley P.

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准确绘制微量样品中单分子复合物 3D 几何结构的能力是一个具有挑战性的目标,它将带来对分子力学的新见解,并为单分子结构蛋白质组学提供一种方法。为了实现这一点,我们开发了一种高分辨率力谱方法,能够测量天然折叠蛋白质复合物中标记位点之间的多个距离。我们的方法将可重新配置的纳米级设备(我们称之为 DNA 纳米开关卡尺)与基于力的条形码系统相结合,以区分每个测量位置。我们通过重建天然折叠的链霉亲和素中生物素结合位点的四面体几何形状来展示我们的方法,与之前报道的结构具有 1.5–2.5 Å 的一致性。
The ability to accurately map the 3D geometry of single-molecule complexes in trace samples is a challenging goal that would lead to new insights into molecular mechanics and provide an approach for single-molecule structural proteomics. To enable this, we have developed a high-resolution force spectroscopy method capable of measuring multiple distances between labeled sites in natively folded protein complexes. Our approach combines reconfigurable nanoscale devices, we call DNA nanoswitch calipers, with a force-based barcoding system to distinguish each measurement location. We demonstrate our approach by reconstructing the tetrahedral geometry of biotin-binding sites in natively folded streptavidin, with 1.5–2.5 Å agreement with previously reported structures.
使用原子力显微镜在蛋白质复合物内进行化学基团的成像和三维重建。
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