非常简单的 TROSY、CRINEPT 和多量子相干组合，可在用于大型蛋白质的 HN(CO)CA 实验中增强信号。

Very simple combination of TROSY, CRINEPT and multiple quantum coherence for signal enhancement in an HN(CO)CA experiment for large proteins.

机构信息

ETH Zurich, Department of Physical Chemistry, Zurich, Switzerland.

出版信息

J Magn Reson. 2011 Apr;209(2):310-4. doi: 10.1016/j.jmr.2011.01.031. Epub 2011 Feb 3.

DOI:10.1016/j.jmr.2011.01.031

Abstract

Sensitivity enhancement in liquid state nuclear magnetic resonance (NMR) triple resonance experiments for the sequential assignment of proteins is important for the investigation of large proteins or protein complexes. We present here the 3D TROSY-MQ/CRINEPT-HN(CO)CA which makes use of a ¹⁵N-¹H-TROSY element and a ¹³C'-¹³CA CRINEPT step combined with a multiple quantum coherence during the ¹³CA evolution period. Because of the introduction of these relaxation-optimized elements and 10 less pulses required, when compared with the conventional TROSY-HN(CO)CA experiment an average signal enhancement of a factor of 1.8 was observed for the membrane protein-detergent complex KcsA with a rotational correlation time τ(c) of around 60 ns.

摘要

在液体状态下的核磁共振（NMR）三重共振实验中，提高对蛋白质的顺序赋值的灵敏度对于研究大型蛋白质或蛋白质复合物非常重要。我们在此介绍了 3D TROSY-MQ/CRINEPT-HN(CO)CA，它利用了 ¹⁵N-¹H-TROSY 元件和 ¹³C'-¹³CA CRINEPT 步骤，并在 ¹³CA 演化期间结合了多个量子相干。由于引入了这些弛豫优化元件，与传统的 TROSY-HN(CO)CA 实验相比，当旋转相关时间 τ(c)约为 60 ns 的膜蛋白-去污剂复合物 KcsA 时，所需的脉冲减少了 10 个，平均信号增强了 1.8 倍。