Synthesis of proteins from [35S]methionine by guinea pig megakaryocytes in vivo and time course of appearance of newly synthesized proteins in platelets.
Synthesis of proteins from [35S]methionine by guinea pig megakaryocytes in vivo and time course of appearance of newly synthesized proteins in platelets.
复制标题
DOI：
10.1089/ten.2005.11.1159
复制
发表时间：
1990-09
期刊：
影响因子：
20.3
通讯作者：
B. Schick
中科院分区：
文献类型：
--
作者：
B. Schick
在豚鼠体内研究了蛋白质合成与巨核细胞成熟的关系。豚鼠注射单剂量[35 S]蛋氨酸。每天分离巨核细胞和血小板，持续4天，通过DEAE-Sephacel色谱法分离来自两种细胞的蛋白质，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和荧光照相法进行分析。在注射后3小时，对与SDS-PAGE凝胶上染色条带对应的巨核细胞中的所有蛋白进行放射性标记。巨核细胞放射性的最大损失发生在注射后1 - 3天。在3小时时仅观察到血小板蛋白的痕量标记，几乎完全代表分子量为47，000、52，000和66，000的三个条带。在24小时时，仅存在约13%的最大标记，但并非所有蛋白质都被标记。最大标记在第3天。血小板在第3天的标记模式与巨核细胞在第3小时的标记模式相同。非巨核细胞骨髓细胞的蛋白质谱不同于血小板和巨核细胞。这里提供的数据表明，巨核细胞中的大多数蛋白质合成至少在血小板释放到循环中之前24小时完成，并且表明在成熟的终末阶段合成的蛋白质中具有某些特异性。
The relationship of protein synthesis to megakaryocyte maturation has been studied in guinea pigs in vivo. Guinea pigs were injected with a single dose of [35S]methionine. Megakaryocytes and platelets were isolated daily for 4 days, and proteins from both cells were isolated by DEAE-Sephacel chromatography and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. All proteins in megakaryocytes corresponding to stained bands on the SDS-PAGE gels were radiolabeled at 3 hours after injection. The greatest loss of radioactivity from the megakaryocytes occurred between 1 and 3 days after injection. Only trace labeling of platelet proteins was seen at 3 hours, representing almost entirely three bands at molecular weights 47,000, 52,000, and 66,000. At 24 hours only about 13% of the maximal labeling was present, but not all proteins were labeled. The maximal labeling was at 3 days. The pattern of labeling of platelets at 3 days was identical to that of megakaryocytes at 3 hours. The protein pattern of nonmegakaryocytic marrow cells was different from that of the platelets and megakaryocytes. Data presented here suggest that most protein synthesis in megakaryocytes is completed at least 24 hours before release of the platelets to the circulation, and suggest some specificity in the proteins that are synthesized at the terminal stages of maturation.