Improving particle quality in cryo-EM analysis using a PEGylation method
Improving particle quality in cryo-EM analysis using a PEGylation method
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使用聚乙二醇化方法提高冷冻电镜分析中的颗粒质量
DOI：
10.1016/j.str.2021.05.004
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发表时间：
2021
期刊：
影响因子：
5.7
通讯作者：
Ohto Umeharu
中科院分区：
文献类型：
--
作者：
Zhang Zhikuan;Shigematsu Hideki;Shimizu Toshiyuki;Ohto Umeharu
冷冻电子显微镜（cryo-EM）是一种广泛应用于结构生物学研究的技术，近年来得到了广泛的发展。然而，其样品玻璃化过程是一个主要的限制，因为它会导致严重的颗粒聚集和/或变性。这种效应被认为是因为在玻璃化过程中颗粒倾向于粘附在“致命的”空气-水界面上。在这里，我们报告了一种聚乙二醇化蛋白质的方法，可以有效地保护颗粒免受玻璃化过程中的这些问题。该方法简化了精细调整玻璃化条件的费力过程，允许分析否则将被丢弃的样品。
Cryo-electron microscopy (cryo-EM) is widely used for structural biology studies and has been developed extensively in recent years. However, its sample vitrification process is a major limitation because it causes severe particle aggregation and/or denaturation. This effect is thought to occur because particles tend to stick to the "deadly" air-water interface during vitrification. Here, we report a method for PEGylation of proteins that can efficiently protect particles against such problems during vitrification. This method alleviates the laborious process of fine-tuning the vitrification conditions, allowing for analysis of samples that would otherwise be discarded.