Water's Variable Role in Protein Stability Uncovered by Liquid-Observed Vapor Exchange NMR.
Water's Variable Role in Protein Stability Uncovered by Liquid-Observed Vapor Exchange NMR.
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DOI：
10.1021/acs.biochem.1c00552
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发表时间：
2021-10-19
期刊：
影响因子：
2.9
通讯作者：
Pielak GJ
中科院分区：
文献类型：
--
作者：
Crilly CJ;Eicher JE;Warmuth O;Atkin JM;Pielak GJ
水对蛋白质的结构和稳定性至关重要，但我们对水如何塑造蛋白质的理解还远远不够。我们对蛋白质-水相互作用的不完整知识部分是由于长期以来的技术无法通过实验评估水的去除如何影响局部蛋白质结构。现在可以通过液体观察蒸汽交换（LOVE）NMR获得脱水蛋白质结构的残留水平信息，这是一种溶液NMR技术，可以量化脱水蛋白质和D2 O蒸汽的未保护酰胺质子之间的氢氘交换程度。在这里，我们应用LOVE NMR，傅里叶变换红外光谱，和溶液氢氘交换到球状蛋白质GB 1，CI 2，和其两个变体连接突变引起的脱水蛋白质结构的变化，在溶液结构和稳定性的变化。我们发现，一个突变，不稳定的GB 1在溶液中不影响其脱水结构，而一个突变，稳定CI 2在溶液中使几个地区的蛋白质更容易脱水诱导展开，这表明水是主要负责的GB 1变体的不稳定，但在CI 2变体中起着稳定的作用。我们的研究结果表明，脱水蛋白质结构的变化不能单独从溶液稳定性的变化来预测，并证明了LOVE NMR揭示水在蛋白质稳定性中的可变作用的能力。LOVE NMR进一步应用于其他蛋白质及其变体将提高预测和调节水合和脱水状态下蛋白质结构和稳定性的能力，以应用于医学和生物技术。
Water is essential to protein structure and stability, yet our understanding of how water shapes proteins is far from thorough. Our incomplete knowledge of protein–water interactions is due in part to a long-standing technological inability to assess experimentally how water removal impacts local protein structure. It is now possible to obtain residue-level information on dehydrated protein structures via liquid-observed vapor exchange (LOVE) NMR, a solution NMR technique that quantifies the extent of hydrogen–deuterium exchange between unprotected amide protons of a dehydrated protein and D2O vapor. Here, we apply LOVE NMR, Fourier transform infrared spectroscopy, and solution hydrogen–deuterium exchange to globular proteins GB1, CI2, and two variants thereof to link mutation-induced changes in the dehydrated protein structure to changes in solution structure and stability. We find that a mutation that destabilizes GB1 in solution does not affect its dehydrated structure, whereas a mutation that stabilizes CI2 in solution makes several regions of the protein more susceptible to dehydration-induced unfolding, suggesting that water is primarily responsible for the destabilization of the GB1 variant but plays a stabilizing role in the CI2 variant. Our results indicate that changes in dehydrated protein structure cannot be predicted from changes in solution stability alone and demonstrate the ability of LOVE NMR to uncover the variable role of water in protein stability. Further application of LOVE NMR to other proteins and their variants will improve the ability to predict and modulate protein structure and stability in both the hydrated and dehydrated states for applications in medicine and biotechnology.
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DOI：
10.1073/pnas.37.4.205
发表时间：
1951-01-01
影响因子：
11.1
作者：
PAULING, L;COREY, RB;BRANSON, HR
通讯作者：
BRANSON, HR
影响因子：
3.9
作者：
Allison, SD;Chang, B;Carpenter, JF
通讯作者：
Carpenter, JF
影响因子：
62.1
作者：
Bellissent-Funel MC;Hassanali A;Havenith M;Henchman R;Pohl P;Sterpone F;van der Spoel D;Xu Y;Garcia AE
通讯作者：
Garcia AE
影响因子：
5.6
作者：
Itzhaki, LS;Neira, JL;Fersht, AR
通讯作者：
Fersht, AR
影响因子：
3.1
作者：
Moorthy, Balakrishnan S.;Iyer, Lavanya K.;Topp, Elizabeth M.
通讯作者：
Topp, Elizabeth M.