Cryo-EM samples of gas-phase purified protein assemblies using native electrospray ion-beam deposition.
Cryo-EM samples of gas-phase purified protein assemblies using native electrospray ion-beam deposition.
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DOI：
10.1039/d2fd00065b
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发表时间：
2022-11-08
影响因子：
3.4
通讯作者：
Rauschenbach, Stephan
中科院分区：
文献类型：
--
作者：
Esser, Tim K.;Bohning, Jan;Fremdling, Paul;Bharat, Tanmay;Gault, Joseph;Rauschenbach, Stephan
越来越多的生物分子功能的研究间接联合收割机质谱（MS）与成像技术，如冷冻电子显微镜（cryo-EM）。这种方法可以获得有关天然蛋白质复合物的均匀性，化学计量，形状和相互作用的信息，与高分辨率蛋白质结构互补。我们最近已经证明了TEM样品制备通过本地电喷雾离子束沉积（ES-IBD）作为本地MS和冷冻EM之间的直接联系。该工作流程形成了一种潜在的新途径，以可靠地制备均匀的冷冻EM样品，并更好地了解原生溶液相和原生气相结构之间的关系。然而，工作流程的许多方面需要理解和优化，以获得与最先进的冷冻EM相当的性能。在这里，我们扩大了以前的讨论的关键因素，通过探测基板类型和沉积能量的影响。我们目前和讨论的显微照片，从原生ES-IBD样品与无定形碳，石墨烯，和氧化石墨烯，以及着陆能量在2和150 eV之间的范围内，每电荷。
An increasing number of studies on biomolecular function indirectly combine mass spectrometry (MS) with imaging techniques such as cryo electron microscopy (cryo-EM). This approach allows information on the homogeneity, stoichiometry, shape, and interactions of native protein complexes to be obtained, complementary to high-resolution protein structures. We have recently demonstrated TEM sample preparation via native electrospray ion-beam deposition (ES-IBD) as a direct link between native MS and cryo-EM. This workflow forms a potential new route to the reliable preparation of homogeneous cryo-EM samples and a better understanding of the relation between native solution-phase and native-like gas-phase structures. However, many aspects of the workflow need to be understood and optimized to obtain performance comparable to that of state-of-the-art cryo-EM. Here, we expand on the previous discussion of key factors by probing the effects of substrate type and deposition energy. We present and discuss micrographs from native ES-IBD samples with amorphous carbon, graphene, and graphene oxide, as well as landing energies in the range between 2 and 150 eV per charge.
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影响因子：
15
作者：
Anggara K;Zhu Y;Delbianco M;Rauschenbach S;Abb S;Seeberger PH;Kern K
通讯作者：
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影响因子：
13.6
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DOI：
10.1073/pnas.1900774116
发表时间：
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影响因子：
11.1
作者：
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