Yuwen Tairan, Huang Rui, Kay Lewis E
Departments of Molecular Genetics, Biochemistry and Chemistry, University of Toronto, Toronto, ON, Canada.
Program in Molecular Structure and Function, Hospital for Sick Children, Toronto, ON, Canada.
J Biomol NMR. 2017 Jul;68(3):215-224. doi: 10.1007/s10858-017-0121-x. Epub 2017 Jun 24.
Although N- and C-based chemical exchange saturation transfer (CEST) experiments have assumed an important role in studies of biomolecular conformational exchange, H CEST experiments are only beginning to emerge. We present a methyl-TROSY H CEST experiment that eliminates deleterious H-H NOE dips so that CEST profiles can be analyzed robustly to extract methyl proton chemical shifts of rare protein conformers. The utility of the experiment, along with a version that is optimized for CHD labeled proteins, is established through studies of exchanging protein systems. A comparison between methyl H CEST and methyl H CPMG approaches is presented to highlight the complementarity of the two experiments.
尽管基于N和C的化学交换饱和转移（CEST）实验在生物分子构象交换研究中发挥了重要作用，但H CEST实验才刚刚开始出现。我们提出了一种甲基-TROSY H CEST实验，该实验消除了有害的H-H NOE凹陷，从而可以对CEST谱进行稳健分析，以提取稀有蛋白质构象体的甲基质子化学位移。通过对交换蛋白系统的研究，确定了该实验以及针对CHD标记蛋白优化的版本的实用性。本文还比较了甲基H CEST和甲基H CPMG方法，以突出这两个实验的互补性。
