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低温软着陆可提高蛋白质复合物的结构保存。

Cryogenic Soft Landing Improves Structural Preservation of Protein Complexes.

机构信息

Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.

Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.

出版信息

Anal Chem. 2023 Oct 10;95(40):15094-15101. doi: 10.1021/acs.analchem.3c03228. Epub 2023 Sep 21.

Abstract

We describe an apparatus for the cryogenic landing of particles from the ion beam of a mass spectrometer onto transmission electron microscope grids for cryo-electron microscopy. This system also allows for the controlled formation of thin films of amorphous ice on the grid surface. We demonstrate that as compared to room temperature landings, the use of this cryogenic landing device greatly improves the structural preservation of deposited protein-protein complexes. Furthermore, landing under cryogenic conditions can increase the diversity of particle orientations, allowing for improved 3D structural interpretation. We conclude that this approach allows for the direct coupling of mass spectrometry with cryo-electron microscopy.

摘要

我们描述了一种将质谱仪离子束中的颗粒低温沉积到透射电子显微镜网格上的装置,用于低温电子显微镜。该系统还允许在网格表面上控制形成非晶冰的薄膜。我们证明,与室温沉积相比,使用这种低温沉积装置可以极大地提高沉积的蛋白质-蛋白质复合物的结构保存。此外,低温沉积条件下可以增加颗粒取向的多样性,从而改善 3D 结构解析。我们得出结论,这种方法可以实现质谱与低温电子显微镜的直接耦合。

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