Cistola D P, Hall K B
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO 63110-1093, USA.
J Biomol NMR. 1995 Jun;5(4):415-9. doi: 10.1007/BF00182285.
A simple approach for detecting internal water molecules in proteins in solution is described. This approach combines 19F-detected heteronuclear Overhauser and exchange spectroscopy (HOESY) with site-specific 19F substitution. The model system employed was intestinal fatty acid-binding protein complexed with [2-mono-19F]-palmitate. An intense cross peak was observed between the fluorine and a buried water molecule, as defined in the 1.98 A crystal structure of the complex. From HOESY spectra, the fluorine-water distance was estimated to be 2.1 A, in agreement with the crystal structure. This approach may be applicable to macromolecules that are too large for 1H-detected NMR methods.
本文描述了一种检测溶液中蛋白质内部水分子的简单方法。该方法将¹⁹F检测的异核Overhauser和交换光谱（HOESY）与位点特异性¹⁹F取代相结合。所采用的模型系统是与[2-单-¹⁹F]-棕榈酸酯复合的肠脂肪酸结合蛋白。在氟与一个埋藏的水分子之间观察到一个强烈的交叉峰，如该复合物1.98 Å晶体结构所定义。从HOESY光谱估计，氟与水的距离为2.1 Å，与晶体结构一致。这种方法可能适用于对¹H检测的核磁共振方法来说太大的大分子。
