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Paramagnetic relaxation enhancement to improve sensitivity of fast NMR methods: application to intrinsically disordered proteins

机译:顺磁性弛豫增强可提高快速NMR方法的灵敏度:应用于内在无序的蛋白质

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摘要

We report enhanced sensitivity NMR measurements of intrinsically disordered proteins in the presence of paramagnetic relaxation enhancement (PRE) agents such as Ni2+-chelated DO2A. In proton-detected 1H-15N SOFAST-HMQC and carbon-detected (H-flip)13CO-15N experiments, faster longitudinal relaxation enables the usage of even shorter interscan delays. This results in higher NMR signal intensities per units of experimental time, without adverse line broadening effects. At 40 mmol·L−1 of the PRE agent, we obtain a 1.7- to 1.9-fold larger signal to noise (S/N) for the respective 2D NMR experiments. High solvent accessibility of intrinsically disordered protein (IDP) residues renders this class of proteins particularly amenable to the outlined approach.
机译:我们报道了在顺磁弛豫增强(PRE)试剂如Ni 2 + -螯合的DO2A存在下,固有紊乱蛋白的增强的NMR灵敏度测量。在质子检测到的 1 H- 15 N SOFAST-HMQC和碳检测到的(H-flip) 13 CO- 15 < / sup> N个实验,更快的纵向松弛使得可以使用更短的扫描间延迟。这导致每单位实验时间更高的NMR信号强度,而没有不利的谱线展宽效果。在40mmol·Lsup-1的PRE试剂中,我们进行的2D NMR实验得到的信噪比(S / N)为1.7-1.9倍。本质上无序的蛋白质(IDP)残基的高溶剂可及性使此类蛋白质特别适合于概述的方法。

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