Chevelkov Veniamin, van Rossum Barth J, Castellani Federica, Rehbein Kristina, Diehl Anne, Hohwy Morten, Steuernagel Stefan, Engelke Frank, Oschkinat Hartmut, Reif Bernd
Institut für Organische Chemie und Biochemie II, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching, Germany.
J Am Chem Soc. 2003 Jul 2;125(26):7788-9. doi: 10.1021/ja029354b.
In this communication, we demonstrate the feasibility of 1H detection in MAS solid-state NMR for a microcrystalline, uniformly 2H,15N-labeled sample of a SH3 domain of chicken alpha-spectrin, using pulsed field gradients for suppression of water magnetization. Today, B0 gradients are employed routinely in solution-state NMR for coherence order selection and solvent suppression. We suggest to use gradients to purge water magnetization which cannot be suppressed using conventional water suppression schemes. The achievable gain in sensitivity for 1H detection is in the order of 5 compared to the 15N detected version of the experiment (at a MAS rotation frequency of 13.5 kHz). We expect that this labeling concept which achieves high sensitivity due to 1H detection, in combination with the possibility to measure long range 1H-1H distances as we have shown previously, to be a useful tool for the determination of protein structures in the solid state.
在本通讯中，我们展示了在MAS固态核磁共振中对鸡α-血影蛋白SH3结构域的微晶、均匀2H、15N标记样品进行1H检测的可行性，使用脉冲场梯度抑制水的磁化。如今，B0梯度在溶液态核磁共振中常用于相干序选择和溶剂抑制。我们建议使用梯度来消除使用传统水抑制方案无法抑制的水磁化。与实验的15N检测版本相比（在13.5 kHz的MAS旋转频率下），1H检测可实现的灵敏度增益约为5倍。我们预计，这种由于1H检测而实现高灵敏度的标记概念，结合我们之前所示的测量长程1H-1H距离的可能性，将成为确定固态蛋白质结构的有用工具。
