Monitoring alkaline transitions of yeast iso-1 cytochrome c at natural isotopic abundance using trimethyllysine as a native NMR probe
Monitoring alkaline transitions of yeast iso-1 cytochrome c at natural isotopic abundance using trimethyllysine as a native NMR probe
复制标题
使用三甲基赖氨酸作为天然 NMR 探针监测天然同位素丰度下酵母 iso-1 细胞色素 c 的碱性转变
DOI：
10.1039/c8cc07605g
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发表时间：
2018
影响因子：
4.9
通讯作者：
Liu Maili
中科院分区：
文献类型：
--
作者：
Sun Peng;Wang Qianwen;Yuan Bin;Zhu Qinjun;Jiang Bin;Li Conggang;Lan Wenxian;Cao Chunyang;Zhang Xu;Liu Maili
光谱重叠使得难以使用NMR来绘制大分子的非均相构象集合的构象分布。在这里，我们应用1H-14 N HSQC实验来监测酵母异-1细胞色素c（ycyt c）在天然同位素丰度的碱性构象转变。通过1H-14 N HSQC实验选择性地检测ycyt c的三甲基化Lys 72，并将其用作探针来追踪ycyt c在碱性条件下的构象转变。发现在碱性条件下，至少有四种不同的构象共存。除了天然的结构，赖氨酸73或赖氨酸79协调的构象和部分未折叠的状态与暴露的血红素进行了观察。这些结果表明，该方法是强大的简化光谱的三甲基化的蛋白质，这使得有可能研究复杂的构象转变的天然提取或化学修饰的三甲基化的蛋白质在天然同位素丰度。
Spectral overlap makes it difficult to use NMR for mapping the conformational profile of heterogeneous conformational ensembles of macromolecules. Here, we apply a 1H–14N HSQC experiment to monitor the alkaline conformational transitions of yeast iso-1 cytochrome c (ycyt c) at natural isotopic abundance. Trimethylated Lys72 of ycyt c is selectively detected by a 1H–14N HSQC experiment, and used as a probe to trace conformational transitions of ycyt c under alkaline conditions. It was found that at least four different conformers of ycyt c coexisted under alkaline conditions. Besides the native structure, Lys73 or Lys79 coordinated conformers and a partially unfolded state with exposed heme were observed. These results indicate that the method is powerful at simplifying spectra of a trimethylated protein, which makes it possible to study complex conformational transitions of naturally extracted or chemically modified trimethylated protein at natural isotopic abundance.