NMR STUDIES OF CELLULAR METABOLISM

本项目旨在开发和应用活体核磁共振技术 研究代谢及其扰动的光谱方法 化学毒素 最近在这一领域的工作重点是分析 关于氟化化合物代谢和使用氟化 化合物来探测细胞功能 的结构确定 代谢物的氟化化合物的NMR提出了一个基本的 问题：19F NMR谱通常只涉及一个或几个 每个化合物的共振，因此没有足够的信息可用于 做一个完整的描述。 或者，质子NMR谱是 很难处理，因为基本上所有存在于 典型的样品如组织提取物具有多个质子共振。 在过去的一年里，我们一直在努力推广一种技术， 最初开发用于表征磷代谢物， 氟化化合物的分析。 这种方法主要涉及 INEPT偏振转移的组合，其仅用于激发 这些质子耦合到氟，与中继极化TOCSY 实验来激发分子中的其他质子。 初步研究 已经集中在应用这种方法来确定 氟化葡萄糖的代谢物。 第二个系列的研究已经 利用氟化葡萄糖作为葡萄糖转运的探针 在红细胞中。 最近的心脏灌注研究表明， 腺苷受体拮抗剂明显干扰葡萄糖 运输 研究了这些拮抗剂中的几种对 氟化葡萄糖通过红细胞膜的转运证实了 这个结论。

This program is aimed at the development and application of in vivo NMR spectroscopic methods for studying metabolism and its perturbation by chemical toxins. Recent efforts in this area have focused on the analysis of the metabolism of fluorinated compounds and on the use of fluorinated compounds to probe cellular function. Determination of the structure of metabolites of fluorinated compounds by NMR presents a fundamental problem: the 19F NMR spectra typically involve only one or a few resonances per compound so that not enough information is available to make a complete characterization. Alternatively, proton NMR spectra are difficult to deal with since essentially all of the molecules present in typical samples such as tissue extracts have multiple proton resonances. During the past year we have worked to extend a technique which we initially developed for the characterization of phosphorous metabolites to the analysis of fluorinated compounds. This approach basically involves a combination of INEPT polarization transfer which is used to excite only those protons coupled to fluorine, with a relayed polarization TOCSY experiment to excite the other protons in the molecule. Initial studies have focused on the application of this method to a determination of the metabolites of fluorinated glucose. A second series of studies has been carried out to utilize fluorinated glucose as a probe of glucose transport in erythrocytes. Recent perfused heart studies indicated that several adenosine receptor antagonists were apparently interfering with glucose transport. Studies of the effects of several of these antagonists on the transport of fluorinated glucose across erythrocyte membranes confirm this conclusion.