Department of Chemistry, University Konstanz, Research School Chemical Biology (KoRS-CB), Universitätsstrasse 10, 78457, Konstanz, Germany.
Chembiochem. 2020 Dec 11;21(24):3575-3579. doi: 10.1002/cbic.202000413. Epub 2020 Sep 16.
In living organisms, protein folding and function take place in an inhomogeneous, highly crowded environment possessing a concentration of diverse macromolecules of up to 400 g/L. It has been shown that the intracellular environment has a pronounced effect on the stability, dynamics and function of the protein under study, and has for this reason to be considered. However, most protein studies neglect the presence of these macromolecules. Consequently, we probe here the overall thermodynamic stability of cold shock protein B from Bacillus subtilis (BsCspB) in cell lysate. We found that an increase in cell lysate concentration causes a monotonic increase in the thermodynamic stability of BsCspB. This result strongly underlines the importance of considering the biological environment when inherent protein parameters are quantitatively determined. Moreover, we demonstrate that targeted application of F NMR spectroscopy operates as an ideal tool for protein studies performed in complex cellular surroundings.
在活生物体中，蛋白质折叠和功能发生在非均相、高度拥挤的环境中，该环境具有高达 400 g/L 的各种大分子的浓度。已经表明，细胞内环境对所研究的蛋白质的稳定性、动力学和功能有显著影响，因此必须加以考虑。然而，大多数蛋白质研究忽略了这些大分子的存在。因此，我们在这里探测枯草芽孢杆菌冷休克蛋白 B（BsCspB）在细胞裂解物中的整体热力学稳定性。我们发现，细胞裂解物浓度的增加会导致 BsCspB 的热力学稳定性呈单调增加。这一结果有力地强调了在定量确定固有蛋白质参数时考虑生物环境的重要性。此外，我们证明，靶向应用 F NMR 光谱学是在复杂细胞环境中进行蛋白质研究的理想工具。
