"Invisible" conformers of an antifungal disulfide protein revealed by constrained cold and heat unfolding, CEST-NMR experiments, and molecular dynamics calculations.
"Invisible" conformers of an antifungal disulfide protein revealed by constrained cold and heat unfolding, CEST-NMR experiments, and molecular dynamics calculations.
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DOI：
10.1002/chem.201404879
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发表时间：
2015-03-23
影响因子：
4.3
通讯作者：
Batta, Gyula
中科院分区：
文献类型：
--
作者：
Fizil, Adam;Gaspari, Zoltan;Barna, Terezia;Marx, Florentine;Batta, Gyula
蛋白质构象间的转换被认为是影响蛋白质功能的一个重要因素。为了支持这一点，在几种情况下，检测到隐藏的动态NMR结构高达百分之几的人口。在这里，我们通过热去折叠的二态和三态分析表明，在298 K下，在富含二硫化物的蛋白质中，隐藏态的数量可能占20- 40%。 此外，灵敏的15 N-CEST NMR实验鉴定了与折叠的PAF蛋白缓慢交换的低填充（0.15%）状态。值得注意的是，其他技术未能识别NMR“暗物质”的其余部分。从实验和分子动力学计算的化学位移的温度依赖性的比较表明，PAF的隐藏构象不同的环和终端区域，是最相似的进化保守的核心。我们的观察指出，存在一个复杂的构象景观与多个构象状态的动态平衡，不同的汇率大概负责相当一部分的完全隐藏的性质。
Transition between conformational states in proteins is being recognized as a possible key factor of function. In support of this, hidden dynamic NMR structures were detected in several cases up to populations of a few percent. Here, we show by two- and three-state analysis of thermal unfolding, that the population of hidden states may weight 20–40 % at 298 K in a disulfide-rich protein. In addition, sensitive 15N-CEST NMR experiments identified a low populated (0.15 %) state that was in slow exchange with the folded PAF protein. Remarkably, other techniques failed to identify the rest of the NMR “dark matter”. Comparison of the temperature dependence of chemical shifts from experiments and molecular dynamics calculations suggests that hidden conformers of PAF differ in the loop and terminal regions and are most similar in the evolutionary conserved core. Our observations point to the existence of a complex conformational landscape with multiple conformational states in dynamic equilibrium, with diverse exchange rates presumably responsible for the completely hidden nature of a considerable fraction.
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DOI：
10.1111/j.1742-4658.2009.07011.x
发表时间：
2009-05
期刊：
The FEBS journal
影响因子：
--
作者：
Batta G;Barna T;Gáspári Z;Sándor S;Kövér KE;Binder U;Sarg B;Kaiserer L;Chhillar AK;Eigentler A;Leiter E;Hegedüs N;Pócsi I;Lindner H;Marx F
通讯作者：
Marx F
影响因子：
3.7
作者：
Cho, Min-Kyu;Xiang, ShengQi;Zweckstetter, Markus
通讯作者：
Zweckstetter, Markus
影响因子：
2.9
作者：
GOOD, NE;WINGET, GD;SINGH, RMM
通讯作者：
SINGH, RMM
影响因子：
13.8
作者：
Csermely, Peter;Palotai, Robin;Nussinov, Ruth
通讯作者：
Nussinov, Ruth
影响因子：
2.9
作者：
CARRA, JH;ANDERSON, EA;PRIVALOV, PL
通讯作者：
PRIVALOV, PL