残余内三重共振实验中的优化路径选择

Optimized pathway selection in intraresidual triple-resonance experiments.

作者信息

Tossavainen Helena, Permi Perttu

机构信息

NMR Laboratory, Structural Biology and Biophysics Program, Institute of Biotechnology, University of Helsinki, P.O. Box 65, FIN-00014, Helsinki, Finland.

出版信息

J Magn Reson. 2004 Oct;170(2):244-51. doi: 10.1016/j.jmr.2004.06.018.

DOI:10.1016/j.jmr.2004.06.018

PMID:15388087

Abstract

An optimized intraresidual pulse sequence element with better sensitivity and suppression of sequential cross peaks is presented. Concatenation of three magnetization transfer delays allows their independent setting, in accordance with the relaxation properties of the individual spins, without concomitantly prolonging the pulse sequence. Additionally, implementations of the scheme to HNCA, HNCACB, and the TROSY based triple-resonance experiments are proposed. The feasibility of the new element was verified by recording HNCA and HNCACB on the small 8.6 kDa protein ubiquitin. The corresponding HNCA-TROSY experiment was tested on a larger protein, the 30.4 kDa Cel6A from the thermophilic soil bacterium Thermobifida fusca at 800 (1)H MHz.

摘要

提出了一种优化的残余内脉冲序列元件，具有更好的灵敏度和对连续交叉峰的抑制能力。三个磁化转移延迟的串联允许根据各个自旋的弛豫特性独立设置它们，而不会相应地延长脉冲序列。此外，还提出了该方案在HNCA、HNCACB和基于TROSY的三共振实验中的实现方法。通过在8.6 kDa的小蛋白质泛素上记录HNCA和HNCACB，验证了新元件的可行性。相应的HNCA-TROSY实验在较大的蛋白质上进行了测试，该蛋白质是来自嗜热土壤细菌嗜热栖热放线菌的30.4 kDa的Cel6A，在800 (1)H MHz下进行。