Protein linewidth and solvent dynamics in frozen solution NMR.
Protein linewidth and solvent dynamics in frozen solution NMR.
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DOI：
10.1371/journal.pone.0047242
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
McDermott AE
中科院分区：
文献类型：
--
作者：
Siemer AB;Huang KY;McDermott AE
冷冻水溶液中蛋白质的固态NMR是结构生物学中潜在的强大技术，特别是如果它与动态核极化信号增强策略相结合。关于低温下冷冻溶液蛋白质样品的NMR研究的一个问题是，它们可能具有较差的线宽，从而阻止高分辨率研究。为了进一步了解溶剂壳的组成和温度如何影响蛋白质的线宽，我们记录了不同温度下冷冻水和冷冻甘油水溶液中泛素的1H，2 H和13 C光谱。我们发现，13 C蛋白质的线宽一般随温度的降低而增加。发现这种谱线增宽是不均匀的，并且与质子退耦无关。在纯水中，我们观察到一个突然的线增宽与冻结的散装溶剂，然后在较低的温度下连续线增宽。在冷冻甘油-水，我们没有观察到一个突然的线加宽和NMR线一般窄于纯水在相同的温度。表征与蛋白质交换的水的动力学的1H和2 H测量表明，13 C线增宽相对独立于各向同性水运动的停滞。
Solid-state NMR of proteins in frozen aqueous solution is a potentially powerful technique in structural biology, especially if it is combined with dynamic nuclear polarization signal enhancement strategies. One concern regarding NMR studies of frozen solution protein samples at low temperatures is that they may have poor linewidths, thus preventing high-resolution studies. To learn more about how the solvent shell composition and temperature affects the protein linewidth, we recorded 1H, 2H, and 13C spectra of ubiquitin in frozen water and frozen glycerol-water solutions at different temperatures. We found that the 13C protein linewidths generally increase with decreasing temperature. This line broadening was found to be inhomogeneous and independent of proton decoupling. In pure water, we observe an abrupt line broadening with the freezing of the bulk solvent, followed by continuous line broadening at lower temperatures. In frozen glycerol-water, we did not observe an abrupt line broadening and the NMR lines were generally narrower than for pure water at the same temperature. 1H and 2H measurements characterizing the dynamics of water that is in exchange with the protein showed that the 13C line broadening is relatively independent from the arrest of isotropic water motions.
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固体和溶液生物NMR的高场动态核极化。
影响因子：
1
作者：
Barnes, A. B.;De Paepe, G.;van der Wel, P. C. A.;Hu, K. -N.;Joo, C. -G.;Bajaj, V. S.;Mak-Jurkauskas, M. L.;Sirigiri, J. R.;Herzfeld, J.;Temkin, R. J.;Griffin, R. G.
通讯作者：
Griffin, R. G.
影响因子：
2.9
作者：
KENNEDY, SD;BRYANT, RG
通讯作者：
BRYANT, RG
影响因子：
3.8
作者：
Ringe, D;Petsko, GA
通讯作者：
Petsko, GA
DOI：
10.1006/jmra.1995.0010
发表时间：
1995-10-01
期刊：
JOURNAL OF MAGNETIC RESONANCE SERIES A
影响因子：
--
作者：
BIELECKI, A;BURUM, DP
通讯作者：
BURUM, DP
影响因子：
15
作者：
Sharpe, S;Kessler, N;Tycko, R
通讯作者：
Tycko, R