Daniel J L, Molish I R, Holmsen H
Biochim Biophys Acta. 1980 Oct 15;632(3):444-53. doi: 10.1016/0304-4165(80)90240-8.
The study of adenine nucleotide metabolism is complicated by cellular nucleotide compartmentalization. In platelets, we have been able to use radioactive labeling with adenine to measure, precisely and accurately, changes in the cytoplasmic/mitochondrial pool of adenine nucleotides inspite of the fact that 60% of adenine nucleotides are present in amine-storing granules. High performance liquid chromatography was used to measure the concentrations of ATP, ADP, AMP and IMP from [14C]adenine-labeled platelets under conditions where the granule pool was absent. The radioactive measurements were directly proportional to the chemical measurements. Ethanol-insoluble, actin-bound ADP also had the same specific radioactivity as other metabolic adenine nucleotides. Since this pool can be directly separated from the bulk of cellular nucleotides, its specific radioactivity can be easily measured and used to calculate the concentration of each cytoplasmic adenine nucleotide from its measured radioactivity. These methods may be applicable to other cells.
腺嘌呤核苷酸代谢的研究因细胞内核苷酸的区室化而变得复杂。在血小板中，尽管60%的腺嘌呤核苷酸存在于胺储存颗粒中，但我们仍能够使用腺嘌呤进行放射性标记，精确且准确地测量腺嘌呤核苷酸的细胞质/线粒体池的变化。在不存在颗粒池的条件下，使用高效液相色谱法测量[14C]腺嘌呤标记的血小板中ATP、ADP、AMP和IMP的浓度。放射性测量与化学测量成正比。乙醇不溶性、肌动蛋白结合的ADP也具有与其他代谢腺嘌呤核苷酸相同的比放射性。由于这个池可以直接与大部分细胞核苷酸分离，其比放射性可以很容易地测量，并用于根据测量的放射性计算每个细胞质腺嘌呤核苷酸的浓度。这些方法可能适用于其他细胞。
