Effect of Buffer on Protein Stability in Aqueous Solutions: A Simple Protein Aggregation Model.
Effect of Buffer on Protein Stability in Aqueous Solutions: A Simple Protein Aggregation Model.
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DOI：
10.1021/acs.jpcb.0c10339
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发表时间：
2021-03-18
期刊：
影响因子：
--
通讯作者：
Hribar-Lee B
中科院分区：
文献类型：
--
作者：
Brudar S;Hribar-Lee B
蛋白质的液-液相分离（LLPS）最近与许多疾病的发病有关。尽管在蛋白质聚集这一领域进行了几项研究，但缓冲特异性效应似乎总是被忽视。本研究通过测定几种溶菌酶缓冲液（MOPS、磷酸盐、HEPES、羧酸盐）在pH 7.0下的云点温度、第二维里系数和相互作用扩散系数，研究了缓冲液对蛋清溶菌酶（HEWL）相稳定性及其蛋白-蛋白相互作用的影响。结果表明，缓冲分子根据其水合作用在蛋白质表面吸附，并调节其静电稳定性。获得的信息被用于扩展最近开发的粗粒度蛋白质模型，以纳入缓冲特异性效应。通过Wertheim微扰理论，该模型定性准确地预测了实验中观察到的所有hhl缓冲溶液的相分离，并进一步使我们能够在实验无法达到的条件下预测蛋白质配方的相稳定性。由于该理论可以直接扩展到包括多个组分，它提供了一个有用的工具来研究拥挤的细胞样系统中的蛋白质聚集。
Liquid–liquid phase separation (LLPS) of proteins has recently been associated with the onset of numerous diseases. Despite several studies in this area of protein aggregation, buffer-specific effects always seem to be overlooked. In this study we investigated the influence of buffers on the phase stability of hen egg-white lysozyme (HEWL) and its respective protein–protein interactions by measuring the cloud point temperature, second virial coefficient, and interaction diffusion coefficient of several HEWL–buffer solutions (MOPS, phosphate, HEPES, cacodylate) at pH 7.0. The results indicate that the buffer molecules, depending on their hydration, adsorb on the protein surface, and modulate their electrostatic stability. The obtained information was used to extend the recently developed coarse-grained protein model to incorporate buffer-specific effects. Treated by Wertheim’s perturbation theory the model qualitatively correctly predicted the experimentally observed phase separation of all investigated HEWL–buffer solutions, and further allowed us to predict the phase stability of protein formulations even in experimentally unattainable conditions. Since the theory can be straightforwardly extended to include multiple components it presents a useful tool to study protein aggregation in crowded cell-like systems.
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DOI：
10.1073/pnas.1917569117
发表时间：
2020-06-16
影响因子：
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