Revealing the complexity of ionic liquid-protein interactions through a multi-technique investigation.
Revealing the complexity of ionic liquid-protein interactions through a multi-technique investigation.
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DOI：
10.1038/s42004-020-0302-5
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发表时间：
2020-05-06
影响因子：
5.9
通讯作者：
Hallett, Jason P.
中科院分区：
文献类型：
--
作者：
Bui-Le, Liem;Clarke, Coby J.;Brohl, Andreas;Brogan, Alex P. S.;Arpino, James A. J.;Polizzi, Karen M.;Hallett, Jason P.
离子液体为生物催化提供了令人兴奋的可能性，因为溶剂的性质为可定制、高能效的生物处理提供了难得的机会。不幸的是，蛋白质和酶在离子液体中通常是不稳定的，人们已经做出了几次尝试来解释为什么；然而，对离子液体-蛋白质相互作用的全面了解仍然难以捉摸。在这里，我们提出了一个分析框架(圆二色谱(CD)、荧光、紫外-可见(UV/VIS)和核磁共振(核磁共振)光谱以及小角X射线散射(SAXS))来探索一种模型蛋白质(绿色荧光蛋白(GFP))在一系列(醋酸盐、氯化物、三氟酸盐)吡咯烷和咪唑盐中的相互作用、结构和稳定性。我们证明，测量蛋白质稳定性需要类似的整体分析框架，而不是提供误导性结论的单一技术评估。我们揭示了特定部位离子液体-蛋白质相互作用的信息，揭示了三氟化钠(最小相互作用的阴离子)诱导蛋白质大小的收缩，从而降低了展开的屏障。像这样强大的框架对于推进非水生物催化和避免与单一技术研究相关的陷阱至关重要。离子液体被用作生物催化中的溶剂，但它们如何与蛋白质的结构相互作用尚不完全清楚。在这里，我们用一套实验技术研究了三种常见的离子液体对绿色荧光蛋白结构的影响，发现了一种任何单一技术都难以捕捉到的复杂关系。
Ionic liquids offer exciting possibilities for biocatalysis as solvent properties provide rare opportunities for customizable, energy-efficient bioprocessing. Unfortunately, proteins and enzymes are generally unstable in ionic liquids and several attempts have been made to explain why; however, a comprehensive understanding of the ionic liquid–protein interactions remains elusive. Here, we present an analytical framework (circular dichroism (CD), fluorescence, ultraviolet-visible (UV/Vis) and nuclear magnetic resonance (NMR) spectroscopies, and small-angle X-ray scattering (SAXS)) to probe the interactions, structure, and stability of a model protein (green fluorescent protein (GFP)) in a range (acetate, chloride, triflate) of pyrrolidinium and imidazolium salts. We demonstrate that measuring protein stability requires a similar holistic analytical framework, as opposed to single-technique assessments that provide misleading conclusions. We reveal information on site-specific ionic liquid–protein interactions, revealing that triflate (the least interacting anion) induces a contraction in the protein size that reduces the barrier to unfolding. Robust frameworks such as this are critical to advancing non-aqueous biocatalysis and avoiding pitfalls associated with single-technique investigations. Ionic liquids are used as solvents in biocatalysis but how they interact with the structures of proteins is imperfectly understood. Here the effect of three common ionic liquids on the structure of green fluorescent protein is studied using a suite of experimental techniques, finding a complex relationship which is poorly captured by any single technique.
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影响因子：
15
作者：
Brogans, Alex P. S.;Halleet, Jason P.
通讯作者：
Halleet, Jason P.
影响因子：
56.9
作者：
Ormo, M;Cubitt, AB;Remington, SJ
通讯作者：
Remington, SJ
影响因子：
8.4
作者：
Brogan, Alex P. S.;Siligardi, Giuliano;Mann, Stephen
通讯作者：
Mann, Stephen
影响因子：
3.3
作者：
Ab Rani, M. A.;Brant, A.;Wilding, R.
通讯作者：
Wilding, R.
影响因子：
3.3
作者：
Nordwald, Erik M.;Kaar, Joel L.
通讯作者：
Kaar, Joel L.