Department of Chemistry, University of Konstanz, Universitätsstrasse. 10, 78457, Konstanz, Germany.
Konstanz Research School Chemical Biology KoRS-CB, University of Konstanz, Universitätsstrasse. 10, 78457, Konstanz, Germany.
Nat Commun. 2020 Nov 13;11(1):5760. doi: 10.1038/s41467-020-19616-w.
The high density of macromolecules affecting proteins due to volume exclusion has been discussed in theory but numerous in vivo experiments cannot be sufficiently understood taking only pure entropic stabilization into account. Here, we show that the thermodynamic stability of a beta barrel protein increases equally at all atomic levels comparing crowded environments with dilute conditions by applying multidimensional high-resolution NMR spectroscopy in a systematic manner. Different crowding agents evoke a pure stabilization cooperatively and do not disturb the surface or integrity of the protein fold. The here developed methodology provides a solid base that can be easily expanded to incorporate e.g. binding partners to recognize functional consequences of crowded conditions. Our results are relevant to research projects targeting soluble proteins in vivo as it can be anticipated that their thermodynamic stability increase comparably and has consequently to be taken into account to coherently understand intracellular processes.
由于体积排阻而影响蛋白质的大分子高密度在理论上已经讨论过了，但仅考虑纯熵稳定的话，许多体内实验并不能得到充分的理解。在这里，我们通过系统地应用多维高分辨率 NMR 光谱学，表明在拥挤环境与稀溶液条件下，β桶状蛋白的热力学稳定性在所有原子水平上都同等增加。不同的拥挤试剂协同地产生纯稳定作用，并且不会干扰蛋白质折叠的表面或完整性。这里开发的方法学提供了一个坚实的基础，可以很容易地扩展到纳入例如结合配偶体，以识别拥挤条件下的功能后果。我们的结果与针对体内可溶性蛋白质的研究项目相关，因为可以预期它们的热力学稳定性会相应增加，因此必须考虑到这一点，以连贯地理解细胞内过程。
