Four-dimensional heteronuclear triple-resonance NMR spectroscopy of interleukin-1 beta in solution.
Four-dimensional heteronuclear triple-resonance NMR spectroscopy of interleukin-1 beta in solution.
复制标题
溶液中白细胞介素 1 β 的四维异核三重共振 NMR 波谱。
作者：
L. Kay;G. Clore;A. Bax;A. Gronenborn
提出了一种通过将蛋白质核磁共振谱的维度增加到四维来显著提高其分辨率的方法。这项技术的威力通过将四维碳-13-氮-15(13C-15N)编辑的核Overhauser效应(NOE)光谱应用于白细胞介素1β(一种由153个残基组成的蛋白质)来证明。NH和脂肪族质子之间的NOE首先通过酰胺15N原子的15N化学位移扩展到第三维，然后通过直接键合的13C原子的13C化学位移扩展到第四维。通过这种方法，完全消除了由于广泛的化学位移重叠和脂肪共振的简并而在NH和脂肪质子之间分配NOE的模糊性，这些NOE仍然存在于三维15N编辑的NOE谱中。因此，从NOE数据中可以获得比迄今可能的更多近似的质子间距离约束，从而将核磁共振三维结构确定的视野扩大到更大的蛋白质。
A method is presented that dramatically improves the resolution of protein nuclear magnetic resonance (NMR) spectra by increasing their dimensionality to four. The power of this technique is demonstrated by the application of four-dimensional carbon-13--nitrogen-15 (13C-15N)--edited nuclear Overhauser effect (NOE) spectroscopy to interleukin-1 beta, a protein of 153 residues. The NOEs between NH and aliphatic protons are first spread out into a third dimension by the 15N chemical shift of the amide 15N atom and subsequently into a fourth dimension by the 13C chemical shift of the directly bonded 13C atoms. By this means ambiguities in the assignment of NOEs between NH and aliphatic protons that are still present in the three-dimensional 15N-edited NOE spectrum due to extensive chemical shift overlap and degeneracy of aliphatic resonances are completely removed. Consequently, many more approximate interproton distance restraints can be obtained from the NOE data than was heretofore possible, thereby expanding the horizons of three-dimensional structure determination by NMR to larger proteins.