Department of NMR-assisted Structural Biology, In-cell NMR Group, Leibniz Institute of Molecular Pharmacology (FMP), Berlin, Germany.
J Biomol NMR. 2011 Dec;51(4):487-95. doi: 10.1007/s10858-011-9577-2. Epub 2011 Oct 19.
We report enhanced sensitivity NMR measurements of intrinsically disordered proteins in the presence of paramagnetic relaxation enhancement (PRE) agents such as Ni(2+)-chelated DO2A. In proton-detected (1)H-(15)N SOFAST-HMQC and carbon-detected (H-flip)(13)CO-(15)N 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.
我们报告了在顺磁弛豫增强（PRE）试剂如 Ni（2+）-螯合 DO2A 存在下，对无规卷曲蛋白质进行增强灵敏度的 NMR 测量。在质子检测（1）H-（15）N SOFAST-HMQC 和碳检测（H-flip）（13）CO-（15）N 实验中，更快的纵向弛豫使我们能够使用更短的扫描间隔。这导致在相同实验时间内，NMR 信号强度更高，而不会产生不利的谱线展宽效应。在 40mmol·L（-1）的 PRE 试剂浓度下，我们获得了相应二维 NMR 实验中信号到噪声（S/N）的 1.7 到 1.9 倍提高。无规卷曲蛋白质（IDP）残基的高溶剂可及性使得这一类蛋白质特别适合于所概述的方法。
