Dynamics of reassembled thioredoxin studied by magic angle spinning NMR: snapshots from different time scales.
Dynamics of reassembled thioredoxin studied by magic angle spinning NMR: snapshots from different time scales.
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通过魔法角度旋转NMR：来自不同时间尺度的快照研究的重新组装硫氧还蛋白的动力学。
DOI：
10.1021/ja9037802
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发表时间：
2009-09-30
影响因子：
15
通讯作者：
Polenova, Tatyana
中科院分区：
文献类型：
--
作者：
Yang, Jun;Tasayco, Maria Luisa;Polenova, Tatyana
固态核磁共振波谱可以在没有整体分子翻滚的情况下用来探测蛋白质内部的动力学。在这项研究中，我们报道了差异富集型1-73(U-13C，15N)/74-108(U-15N)重组硫氧还蛋白在多个时间尺度上的15N骨架动力学，使用了一系列2D和3D MAS核磁共振实验，探讨了骨架酰胺15N的纵向驰豫，1H-15N偶极序参数，15N化学位移各向异性(CSA)，以及依赖温度的和1H T2‘过滤的NCA实验中的信号强度。自旋晶格驰豫速率R_1(R_1=1/T_1)在0.012~0.64 S~(-1)范围内观察到，表明动力学在皮秒到纳秒的尺度上有很大的变化。1H-15N偶极序参数<S>和15N CsA各向异性δσ揭示了重组硫氧还蛋白的骨架迁移率，平均值分别为<S>=0.89±0.06和δσ=92.3±5.2ppm。从不同动力学方法的实验数据集合中，提出了不同时间尺度上的运动之间的某种程度的相关性。对这些固体核磁共振实验得到的动力学参数的分析表明，组成不规则二级结构元素的残基比位于α-螺旋和β-折叠中的残基具有更高的迁移率，α-螺旋和β-折叠残基在动力学上没有明显的系统差异。值得注意的是，由固体核磁共振测量得到的偶极有序参数和相应的溶液核磁共振广义有序参数随着残基数目的变化呈现出类似的定性趋势。通过固体动力学参数与晶体B因子的比较，确定了静态无序对B因子的贡献。本研究中采用的纵向松弛、偶极序参数和CSA线形分析相结合的方法提供了动力学快照，并对这些运动在多个时间尺度上的相关性提供了新的见解。
Solid-state NMR spectroscopy can be used to probe internal protein dynamics in the absence of the overall molecular tumbling. In this study, we report 15N backbone dynamics in differentially enriched 1-73(U-13C, 15N)/74-108(U-15N) reassembled thioredoxin on multiple timescales using a series of 2D and 3D MAS NMR experiments probing the backbone amide 15N longitudinal relaxation, 1H-15N dipolar order parameters, 15N chemical shift anisotropy (CSA), and signal intensities in the temperature-dependent and 1H T2′ -filtered NCA experiments. The spin-lattice relaxation rates R1(R1 = 1/T1) were observed in the range from 0.012 to 0.64 s-1 indicating large site-to-site variations in dynamics on pico- to nanosecond time scales. The 1H-15N dipolar order parameters, <S>, and 15N CSA anisotropies, δσ reveal the backbone mobilities in reassembled thioredoxin, as reflected in the average <S>= 0.89 ± 0.06 and δσ = 92.3 ± 5.2 ppm, respectively. From the aggregate of experimental data from different dynamics methods, some degree of correlation between the motions on the different time scales has been suggested. Analysis of the dynamics parameters derived from these solid-state NMR experiments indicates higher mobilities for the residues constituting irregular secondary structure elements than for those located in the α-helices and β-sheets, with no apparent systematic differences in dynamics between the α-helical and β-sheet residues. Remarkably, the dipolar order parameters derived from the solid-state NMR measurements and the corresponding solution NMR generalized order parameters display similar qualitative trends as a function of the residue number. The comparison of the solid-state dynamics parameters to the crystallographic B-factors has identified the contribution of static disorder to the B-factors. The combination of longitudinal relaxation, dipolar order parameter, and CSA line shape analyses employed in this study provides snapshots of dynamics and a new insight on the correlation of these motions on multiple time scales.
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DOI：
10.1073/pnas.0509133103
发表时间：
2006-03-14
影响因子：
11.1
作者：
Bollen, YJM;Kamphuis, MB;van Mierlo, CPM
通讯作者：
van Mierlo, CPM
影响因子：
4.4
作者：
BENNETT, AE;RIENSTRA, CM;GRIFFIN, RG
通讯作者：
GRIFFIN, RG
影响因子：
1.7
作者：
Baldus, M;Petkova, AT;Griffin, RG
通讯作者：
Griffin, RG
影响因子：
15
作者：
Giraud, N;Blackledge, M;Emsley, L
通讯作者：
Emsley, L
影响因子：
4.4
作者：
Chan, JCC;Tycko, R
通讯作者：
Tycko, R