Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Straße 10, 13125, Berlin, Germany.
J Biomol NMR. 2012 Jan;52(1):31-9. doi: 10.1007/s10858-011-9591-4. Epub 2011 Dec 4.
Biological solid-state nuclear magnetic resonance spectroscopy developed rapidly in the past two decades and emerged as an important tool for structural biology. Resonance assignment is an essential prerequisite for structure determination and the characterization of motional properties of a molecule. Experiments, which rely on carbon or nitrogen detection, suffer, however, from low sensitivity. Recently, we introduced the RAP (Reduced Adjoining Protonation) labeling scheme, which allows to detect backbone and sidechain protons with high sensitivity and resolution. We present here a (1)H-detected 3D (H)CCH experiment for assignment of backbone and sidechain proton resonances. Resolution is significantly improved by employing simultaneous (13)CO and (13)Cβ J-decoupling during evolution of the (13)Cα chemical shift. In total, ~90% of the (1)Hα-(13)Cα backbone resonances of chicken α-spectrin SH3 could be assigned.
在过去的二十年中，生物固态核磁共振波谱学发展迅速，成为结构生物学的重要工具。共振分配是确定结构和分子动态特性的重要前提。然而，依赖于碳或氮检测的实验灵敏度较低。最近，我们引入了 RAP（减少毗邻质子化）标记方案，该方案允许以高灵敏度和分辨率检测骨架和侧链质子。我们在这里提出了一种用于分配骨架和侧链质子共振的（1）H 检测 3D（H）CCH 实验。通过在（13）Cα化学位移演化过程中同时使用（13）CO 和（13）Cβ J 去耦，分辨率得到显著提高。总共，可以分配约 90%的鸡α- spectrin SH3 的（1）Hα-（13）Cα 骨架共振。
