Tauskela J S, Shoubridge E A
McGill University, Montreal Neurological Institute, Canada.
Biochim Biophys Acta. 1993 Oct 3;1158(2):155-65. doi: 10.1016/0304-4165(93)90009-w.
Double quantum filtered (DQF) 23Na-NMR signals were evaluated as a function of [Na+] at constant temperature in two model systems (bovine serum albumin (BSA) and Ficoll 400) and in human red blood cells (RBCs). In model systems, the ratio of double quantum filtered to single quantum (SQ) signal intensities was independent of [Na+], even over a wide range of Na+/K+ ratios. Varying the DQF preparation time affected only the DQF signal intensity. In contrast, in human red blood cells (RBCs) the shape and phase of the DQF intracellular Na+ signal (Na+in) varied as a function of preparation time. Similar observations in cartilage [Eliav, U., Shinar, H. and Navon, G. (1992) J. Magn. Reson. 28, 223-229] have been attributed to the generation of a second- and a third-rank tensor by the DQF pulse sequence, resulting from Na+ ion ordering. By using a DQF sequence which isolates the second-rank tensor only, this component was found to originate from the intracellular Na+ ion pool in human RBCs, as well as from interactions of Na+ ions with the extracellular face of the plasma membrane. The residual quadrupolar splitting for the signal originating from the former environment was shown to be less than the SQ linewidth, explaining its absence in SQ spectra, and this was confirmed by two-dimensional DQF 23Na-NMR experiments. By isolating the contribution from the third-rank tensor exclusively, the ratio of DQF:SQ signal intensities for Na+in in human RBCs was shown to be constant over a 4-fold change in [Na+in] produced by addition of an ionophore (nystatin). This indicates that such changes in physiological state do not alter the efficiency of DQF signal generation in human RBCs.
在两个模型系统(牛血清白蛋白(BSA)和聚蔗糖400)以及人类红细胞(RBC)中,在恒定温度下,双量子滤波(DQF)23Na-NMR信号作为[Na+]的函数进行了评估。在模型系统中,双量子滤波信号强度与单量子(SQ)信号强度的比值与[Na+]无关,即使在很宽的Na+/K+比值范围内也是如此。改变DQF制备时间仅影响DQF信号强度。相比之下,在人类红细胞(RBC)中,DQF细胞内Na+信号(Na+in)的形状和相位随制备时间而变化。软骨中的类似观察结果[埃利亚夫,U.,希纳尔,H.和纳冯,G.(1992年)《磁共振杂志》28卷,223 - 229页]归因于DQF脉冲序列产生的二阶和三阶张量,这是由Na+离子有序排列导致的。通过使用仅分离二阶张量的DQF序列,发现该成分源自人类RBC中的细胞内Na+离子池,以及Na+离子与质膜细胞外表面的相互作用。源自前一种环境的信号的剩余四极分裂显示小于SQ线宽,这解释了其在SQ光谱中不存在的原因,二维DQF 23Na-NMR实验证实了这一点。通过专门分离三阶张量的贡献,发现在添加离子载体(制霉菌素)使[Na+in]变化4倍的情况下,人类RBC中Na+in的DQF:SQ信号强度比值保持恒定。这表明这种生理状态的变化不会改变人类RBC中DQF信号产生的效率。