PHOSPHATE ANALOG PROBES OF RED CELL METABOLISM

磷酸盐类似物是化学上与正磷酸盐有关的化合物，但 对一个或两个氧原子进行化学取代。这些 取代导致原子结构、原子数和原子数的独特变化 可滴定质子的酸性，以及核磁共振中的酸度 每种化合物的光谱。这些独特的特点将被利用 用核磁共振波谱研究人红细胞的两个区域 生理学：膜负离子转运和能量代谢。负离子 运输的研究将通过同时观察细胞内和 胞外共振，或者是由于pH自然发生的 敏感的化学位移，或通过添加一种 胞外顺磁移动剂。膜的快速转运 交换化合物将用饱和转移方法进行研究。 将研究浓度和pH依赖的传输机制， 净阴离子电导将使用电离团和抑制剂进行测量。 阴离子转运蛋白。这些运输特征将是 与阴离子传输的可滴定电荷模型进行了比较。因为这些 化合物具有与正磷酸盐相似的化学成分，它们对 将通过监测血糖来研究红细胞能量代谢。 利用、乳酸生产、细胞内pH和有机合成 磷酸盐(2，3-DPG和ATP)。磷酸类似物对核苷的影响 (肌苷)能量代谢也将被阐明。此外， 类似物对磷酸果糖激酶(一种主要活性物质)活性的影响 正磷酸刺激的糖酵解控制酶)和甘油醛 磷酸脱氢酶(唯一结合磷酸盐的糖酵解酶 从新进入代谢中间体)将被研究。在代谢过程中 将寻找磷酸盐酶解的研究证据 从F-和F-合成氟磷酸盐的类似物 正磷酸盐，以及在酶作用下将磷酸盐类似物掺入 糖酵解代谢物。

Phosphate analogues are compounds chemically related to orthophosphate, but having chemical substitutions for one or two oxygen atoms. These substitutions result in unique changes in atomic structure, the number and acidity of titratible protons, and in the nuclear magnetic resonance (NMR) spectrum of each compound. These unique characteristics will be exploited to study by means of NMR spectroscopy two areas of human erythrocyte physiology: membrane anion transport and energy metabolism. Anion transport will be studied by simultaneous observation of intracellular and extracellular resonances, either naturally occurring as a result of pH sensitive chemical shifts, or induced by means of addition of an extracellular paramagnetic shift reagent. Membrane transport of rapidly exchanging compounds will be studied by saturation-transfer methods. Concentration and pH dependence of the transport mechanism will be studied, and net anion conductance will be measured using ionophores and inhibitors of the anion transport protein. These transport characteristics will be compared to the titratible charge model for anion transport. Since these compounds bear chemical similarities to orthophosphate, their effects on red cell energy metabolism will be studied, by monitoring glucose utilization, lactate production, intracellular pH, and synthesis of organic phosphates (2,3-DPG and ATP). Phosphate analogue effects on nucleoside (inosine) energy metabolism will also be elucidated. In addition, the influence of analogues on the activity of phosphofructokinase (a major glycolysis control enzyme stimulated by orthophosphate) and glyceraldehyde phosphate dehydrogenase (the only glycolytic enzyme incorporating phosphate de novo into metabolic intermediates) will be studied. During metabolic studies evidence wil be sought for enzymatic hydrolysis of phosphate analogues, intracellular synthesis of fluorophosphate from F- and orthophosphate, and for enzymatic incorporation of phosphate analogues into glycolytic metabolites.