Akbey Ümit
Radboud University, Magnetic Resonance Research Center, Institute for Molecules and Materials, Nijmegen, The Netherlands.
Department of Structural Biology, University of Pittsburgh School of Medicine, 3501 Fifth Ave., Pittsburgh, PA, USA.
J Biomol NMR. 2022 Apr;76(1-2):23-28. doi: 10.1007/s10858-021-00388-4. Epub 2022 Jan 8.
Determination of protein structure and dynamics is key to understand the mechanism of protein action. Perdeuterated proteins have been used to obtain high resolution/sensitivty NMR experiments via proton-detection. These methods utilizes H, C and N nuclei for chemical shift dispersion or relaxation probes, despite the existing abundant deuterons. However, a high-sensitivity NMR method to utilize deuterons and e.g. determine site-specific deuterium quadrupolar pattern information has been lacking due to technical difficulties associated with deuterium's large quadrupolar couplings. Here, we present a novel deuterium-excited and proton-detected three-dimensional H-C-H MAS NMR experiment to utilize deuterons and to obtain site-specific methyl H quadrupolar patterns on detuterated proteins for the first time. A high-resolution fingerprint H-N HSQC-spectrum is correlated with the anisotropic deuterium quadrupolar tensor in the third dimension. Results from a model perdeuterated protein has been shown.
确定蛋白质的结构和动力学是理解蛋白质作用机制的关键。全氘代蛋白质已被用于通过质子检测获得高分辨率/灵敏度的核磁共振实验。尽管存在大量的氘核，但这些方法利用氢、碳和氮原子核进行化学位移分散或弛豫探测。然而，由于与氘的大四极耦合相关的技术困难，一种利用氘核并例如确定位点特异性氘四极模式信息的高灵敏度核磁共振方法一直缺乏。在此，我们提出了一种新颖的氘激发和质子检测的三维H-C-H MAS NMR实验，以首次利用氘核并在氘代蛋白质上获得位点特异性甲基氢四极模式。高分辨率指纹H-N HSQC谱在第三维与各向异性氘四极张量相关。已展示了来自模型全氘代蛋白质的结果。
