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

开发具有超高灵敏度的^31P-NMR探针并分析与信号转导相关的磷酸结合蛋白

基本信息

批准号：
13558080
负责人：
TANOKURA Masaru
金额：
$ 9.02万
依托单位：
THE UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

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.

虽然~ 13 <31>P核磁共振（nuclear magnetic resonance，NMR）的灵敏度较高，而且磷酸盐在生物体内主要以ATP等形式存在，<31>但由于~ 13 P的T_2和T_1分别很短和很长，很难获得有用的数据。因此，当推进用于进行实验以检测来自包含单细胞的样品的信号的研究时，迄今为止获得的NMR测量的灵敏度已经成为严重的障碍。然而，近年来，用于NMR测量的磁场强度上升，探针技术也得到改进，通过1H-NMR测量和分析单细胞的可能<31>性也增加了。我们开发了探针设备和用于检测的细胞，可以在生理环境条件下直接监测单细胞。该探头可以改善测量脉冲和接收机的动态范围，使其能够对短T_2作出响应。因此，设备将细胞和组织保持在其长时间存活的状态，这使得测量11 P-NMR光谱成为可能<31>。此外，它还分析了牛奶，这是在该探针中用作检测候选的活物质的混合物。在不进行预处理的情况下，除了进行^1D ^ P NMR测量外，还应用了2D ^1H-1 P NMR光谱，成功地确定了磷酸盐化合物是牛奶中的一种成分<31><31>。