Danio rerio Oocytes for Eukaryotic In-Cell NMR
Danio rerio Oocytes for Eukaryotic In-Cell NMR
复制标题
用于真核细胞内 NMR 的斑马鱼卵母细胞
DOI：
10.1021/acs.biochem.0c00922
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发表时间：
2021
期刊：
影响因子：
2.9
通讯作者：
Pielak, Gary J.
中科院分区：
文献类型：
--
作者：
Thole, Joseph F.;Fadero, Tanner C.;Bonin, Jeffrey P.;Stadmiller, Samantha S.;Giudice, Jonathan A.;Pielak, Gary J.
了解拥挤和复杂的细胞环境如何影响蛋白质的稳定性和动力学，直到最近才成为可能，通过使用细胞内核磁共振等技术。然而，稳定和不稳定相互作用的结合使得简单的预测变得困难。在这里，我们展示了danio卵母细胞作为细胞内核磁共振模型的潜力，该模型可以广泛用于测量蛋白质的稳定性和动力学。我们证明，在真核卵母细胞中，其拥挤程度比其他细胞类型低3 - 6倍，与稀释缓冲液的影响相比，吸引的化学相互作用仍然对蛋白质稳定性和缓慢翻滚时间起主导作用。
Understanding how the crowded and complex cellular milieu affects protein stability and dynamics has only recently become possible by using techniques such as in-cell nuclear magnetic resonance. However, the combination of stabilizing and destabilizing interactions makes simple predictions difficult. Here we show the potential ofDanio reriooocytes as an in-cell nuclear magnetic resonance model that can be widely used to measure protein stability and dynamics. We demonstrate that in eukaryotic oocytes, which are 3–6-fold less crowded than other cell types, attractive chemical interactions still dominate effects on protein stability and slow tumbling times, compared to the effects of dilute buffer.