Structure ( IF 4.4 ) Pub Date : 2021-05-27 , DOI: 10.1016/j.str.2021.05.004 Zhikuan Zhang 1 , Hideki Shigematsu 2 , Toshiyuki Shimizu 1 , Umeharu Ohto 1
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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.
中文翻译：
使用聚乙二醇化方法提高冷冻电镜分析中的颗粒质量
冷冻电子显微镜 (cryo-EM) 广泛用于结构生物学研究，近年来得到了广泛的发展。然而，它的样品玻璃化过程是一个主要限制，因为它会导致严重的粒子聚集和/或变性。这种效应被认为是因为颗粒在玻璃化过程中往往会粘在“致命”的空气-水界面上。在这里，我们报告了一种蛋白质聚乙二醇化的方法，该方法可以有效地保护颗粒在玻璃化过程中免受此类问题的影响。这种方法减轻了微调玻璃化条件的繁琐过程，允许分析否则会被丢弃的样品。
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