The trehalose coating effect on the internal protein dynamics.
The trehalose coating effect on the internal protein dynamics.
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海藻糖涂层对内部蛋白质动力学的影响
DOI：
10.1039/c2cp23098d
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Krushelnitsky
中科院分区：
文献类型：
--
作者：
Hackel;Zinkevich;Belton;Achilles;Reichert;Krushelnitsky
用~(15)N和~(13)C核磁共振实验对冻干粉中的冷休克蛋白(CSP)和海藻糖玻璃基质中的蛋白质进行了比较研究。对这两个样本进行了不同程度的再水化研究。所使用的实验(测量松弛速率R1和R1ρ、动量平均偶极耦合和固体交换法检测化学位移各向异性张量的重定向)允许在从纳秒到秒的相关时间范围内获得关于蛋白质结构特征和内部运动的丰富信息。主要结果如下：(A)与脱水冻干粉末相比，海藻糖涂层使蛋白质结构更加天然，但海藻糖仍然无法消除脱水蛋白质中存在的所有非天然氢键；(B)海藻糖对内部动力学有显著影响：主链N-H基团在纳秒和微秒时间尺度上的运动变慢，而运动幅度保持不变；(C)向CSP-海藻糖混合物中加水后，水分子在蛋白质周围积累，在蛋白质表面和海藻糖基质之间形成一层。(D)蛋白质中NH基团和CH(CH2)基团的动力学水化反应有质的不同：随着蛋白质水化程度的增加，N-H运动的关联时间变短，幅度保持稳定，而CH(CH2)基团的运动幅度增大，关联时间不变。这可以用NH基团和CH(CH2)基团形成氢键的不同能力来解释。
15N and 13C NMR experiments were applied to conduct a comparative study of a cold shock protein (Csp) in two states—lyophilized powder and a protein embedded in a glassy trehalose matrix. Both samples were studied at various levels of rehydration. The experiments used (measuring relaxation rates R1 and R1ρ, motionally averaged dipolar couplings and solid state exchange method detecting reorientation of the chemical shift anisotropy tensor) allow obtaining abundant information on the protein structural features and internal motions in a range of correlation times from nanoseconds to seconds. The main results are: (a) the trehalose coating makes the protein structure more native in comparison with the dehydrated lyophilized powder, however, trehalose still cannot remove all non-native hydrogen bonds which are present in a dehydrated protein; (b) trehalose has an appreciable effect on the internal dynamics: the motion of the backbone N–H groups in the nanosecond and microsecond time scales becomes slower while the motional amplitude remains constant; (c) upon adding water to the Csp–trehalose mixture, water molecules accumulate around proteins forming a layer between the protein surface and the trehalose matrix. The protein dynamics become faster, however, not as fast as in the fully hydrated state; (d) the hydration response of dynamics of the NH and CH(CH2) groups in a protein is qualitatively different: upon increasing protein hydration, the correlation times of the N–H motions become shorter and the amplitude remains stable, and for CH(CH2) groups the motional amplitude increases and the correlation times do not change. This can be explained by a different ability of the NH and CH(CH2) groups to form hydrogen bonds.
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通过 13C 和 1H 固态 NMR 弛豫探测溶菌酶内部动力学对水合的响应
影响因子：
1
作者：
A. Krushelnitsky;D. Reichert
通讯作者：
D. Reichert
在没有中间体的情况下极其快速的蛋白质折叠
DOI：
--
发表时间：
1995
期刊：
Nature Structural Biology
影响因子：
--
作者：
T. Schindler;M. Herrler;M. Marahiel;F. Schmid
通讯作者：
F. Schmid
使用动态蒸汽吸附方法表征无定形海藻糖的吸水性。
影响因子：
3.1
作者：
N. Hunter;C. Frampton;D. Craig;P. Belton
通讯作者：
P. Belton
影响因子：
3.7
作者：
Separovic, F;Lam, YH;Chan, HK
通讯作者：
Chan, HK
通过 15N R(1rho) 位点特异性 NMR 弛豫率研究固体蛋白质中微秒级的迁移率。
影响因子：
15
作者：
A. Krushelnitsky;T. Zinkevich;D. Reichert;Veniamin Chevelkov;B. Reif
通讯作者：
B. Reif