Donohue T M, Sorrell J H, Sorrell M F, Tuma D J
Alcohol Clin Exp Res. 1985 Dec;9(6):526-30. doi: 10.1111/j.1530-0277.1985.tb05597.x.
We investigated the utility of [3H]puromycin as an alternate and adjunct precursor to amino acids for measuring protein synthetic activity in rat liver slices. Slices were incubated in the presence of either [3H]puromycin or radiolabeled valine to compare the incorporation of these isotopic precursors into nascent hepatocellular proteins. Compared to liver slices from controls, comparable decreases in the incorporation of both [3H]puromycin and labeled valine were observed in experiments using slices from fasted rats and in slices preincubated with 25 mM ethanol. Radiolabeling of nascent polypeptides with either [3H]puromycin or labeled valine in liver slices from rats fed a liquid diet containing ethanol was also decreased compared to slices from pair-fed control and chow-fed animals. Our results demonstrated the validity of using [3H]puromycin to detect changes in protein synthetic activity under these conditions. The potential advantage of using [3H]puromycin for in vivo studies is discussed.
我们研究了[3H]嘌呤霉素作为氨基酸的替代和辅助前体用于测量大鼠肝切片中蛋白质合成活性的效用。将切片在[3H]嘌呤霉素或放射性标记的缬氨酸存在下孵育，以比较这些同位素前体掺入新生肝细胞蛋白质中的情况。与对照组的肝切片相比，在使用禁食大鼠的切片以及用25 mM乙醇预孵育的切片进行的实验中，观察到[3H]嘌呤霉素和标记缬氨酸的掺入量均有类似程度的降低。与成对喂养的对照动物和正常饮食喂养动物的切片相比，用含乙醇的流质饮食喂养的大鼠肝切片中，[3H]嘌呤霉素或标记缬氨酸对新生多肽的放射性标记也减少了。我们的结果证明了在这些条件下使用[3H]嘌呤霉素检测蛋白质合成活性变化的有效性。文中还讨论了使用[3H]嘌呤霉素进行体内研究的潜在优势。
