Development of the ^<31>P-NMR Probe with ultra high sensitivity and analysis of phosphate binding proteins, related to signal transduction

虽然^<31>P- nmr（核磁共振）的灵敏度较高，磷酸盐在生物体中主要以ATP等形式存在，但由于T_2和T_1分别很短和很长，^<31>P很难获得有用的数据。因此，迄今为止获得的核磁共振测量的灵敏度一直是推进一项研究的严重障碍，该研究进行了一项实验，以检测含有单细胞的样品的信号。然而，近年来用于核磁共振测量的磁场强度增加，探针技术也得到了改进，用^<31>P-NMR对单细胞进行测量和分析的可能性也增加了。我们研制了探测设备和检测细胞，可以在一定的生理环境下对单细胞进行直接监测。该探头可以改善测量脉冲和接收机的动态范围，使其能够响应短T_2。因此，设备将细胞和组织保持在它们长期存在的状态，这使得测量^<31>的P-NMR光谱成为可能。此外，还对作为探针候选检测物的活性物质混合物的牛奶进行了分析。在不进行预处理的情况下，通过应用2D ^1H-^<31>P核磁共振光谱以及^1D ^<31>P核磁共振测量，成功地将磷酸盐化合物指定为牛奶中的成分。

Although the sensitivity of ^<31>P-NMR (nuclear magnetic resonance) is a comparatively high and phosphate is existed mostly in forms, such as ATP, in the living body, ^<31>P is difficult for obtaining useful data because T_2 and T_1 are very short and long, respectively. Therefore, the sensitivity of the NMR measurement obtained so far has been a serious obstacle when advancing a research for performing an experiment to detect the signals from a sample containing single cells. However, the strength of the magnetic field used for NMR measurement goes up and probe technology was also improved in recent years, a possibility of measuring and analyzing by ^<31>P-NMR the single cells has also increased.We have developed the probe equipment and the cell for detecting can carry out a direct monitoring the single cells in a condition making physiological environment. This probe can be improving a pulse for measurement and a receiver's dynamic range so that it can respond to short T_2 especially. Thus equipment hold the cell and the tissue in the state where it lived for a long time, it made it possible to measure the ^<31>P-NMR spectra. Moreover, it analyzed also about the milk, which is the mixture of the living substances used as the candidate for detection in this probe. It succeeded in assigning the phosphate compound as an ingredient in milk by applying a 2D ^1H-^<31>P NMR spectra in addition to ^1D ^<31>P NMR measurement, without pretreating.