Protein Engineering Network Centers of Excellence and Departments of Molecular and Medical Genetics, Biochemistry and Chemistry, University of Toronto, Toronto, ON, Canada, M5S 1A8.
J Biomol NMR. 1999 Jan;13(1):3-10. doi: 10.1023/A:1008329230975.
A pulse scheme resulting in improved sensitivity in TROSY-based 1HN-detected triple resonance experiments is presented. The approach minimizes relaxation losses which occur during the transfer of transverse magnetization from 15N to 1HN immediately prior to detection. The utility of the method is demonstrated on a complex of methyl protonated, highly deuterated maltose binding protein (MBP, 370 residues) and β- cyclodextrin. Sensitivity gains relative to previous TROSY schemes of approximately 10 and 20% are noted in HNCO spectra of MBP recorded at 25 and 5 °C, respectively, corresponding to molecular correlation times of 23 and 46 ns.
提出了一种脉冲方案，可提高基于 TROSY 的 1HN 检测三重共振实验的灵敏度。该方法最大限度地减少了在检测前立即将横向磁化从 15N 转移到 1HN 时发生的弛豫损失。该方法在甲基质子化、高度氘化麦芽糖结合蛋白（MBP，370 个残基）和β-环糊精复合物上的实验中得到了验证。在分别记录于 25°C 和 5°C 时的 MBP 的 HNCO 谱中，与先前的 TROSY 方案相比，灵敏度分别提高了约 10%和 20%，这对应于分子相关时间为 23 和 46 ns。
