Specific protein synthesis in isolated rat testis leydig cells. Influence of luteinizing hormone and cycloheximide.
Specific protein synthesis in isolated rat testis leydig cells. Influence of luteinizing hormone and cycloheximide.
复制标题
分离的大鼠睾丸间质细胞中的特异性蛋白质合成。
DOI：
--
复制
发表时间：
1977
影响因子：
4.1
通讯作者：
H. van der Molen
中科院分区：
文献类型：
--
作者：
F. Janszen;B. Cooke;H. van der Molen
本文研究了促黄体激素和放线菌酮对大鼠睾丸间质细胞特异性蛋白质合成的影响。在体外与Leydig细胞悬浮液孵育期间，用[114 C]亮氨酸、[3 H]亮氨酸或[35 S]蛋氨酸标记蛋白质。从细胞中提取总蛋白，并通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳分离。用促黄体生成素孵育间质细胞1小时后，未观察到特异性蛋白质合成的可检测增加。然而，在2小时孵育期后，在具有表观mol. wt. 21000（称为“蛋白质21”）。当用[35 S]-甲硫氨酸标记该蛋白质后，将Leydig细胞与放线菌酮再孵育30分钟时，未观察到该蛋白质条带的放射性降低，表明其半衰期不短。然而，另一个蛋白带被检测到，与放线菌酮孵育后迅速消失，反应遵循一级动力学，半衰期约为11分钟。的33000（称为“蛋白33”），被发现位于颗粒部分的Leydig细胞，并不能证明在其他大鼠睾丸细胞类型或血细胞。未观察到促黄体激素对蛋白质33的分子量、亚细胞定位或半衰期的影响。蛋白质33和蛋白质21在促黄体激素对睾丸间质细胞睾酮产生的作用机制进行了讨论。
The effect of luteinizing hormone (luteotropin) and cycloheximide on specific protein synthesis in rat testis Leydig cells has been investigated. Proteins were labelled with either I114C]leucine, [3H]leucine or [35S]methionine during incubation with Leydig-cell suspensions in vitro. Total protein was extracted from the cells and separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. No detectable increase in the synthesis of specific proteins could be observed after incubation of Leydig cells with luteinizing hormone for up to 1 h. However, after a 2h incubation period, an increase in [35S]methionine incorporation was observed in a protein with an apparent mol.wt. of 21000 (referred to as 'protein 21"). When, after labelling of this protein with [35S]-methionine, Leydig cells were incubated for another 30min with cycloheximide, no decrease in radioactivity of this protein band was observed, indicating that it does not have a short half-life. However, another protein band was detected, which after incubation with cycloheximide disappeared rapidly, the reaction following first-order kinetics, with a half-life of about 11 min. This protein, with an apparent mol.wt. of 33000 (referred to as "protein 33"), was found to be located in the particulate fraction of the Leydig cell, and could not be demonstrated in other rat testis-cell types or blood cells. No effect of luteinizing hormone on molecular weight, subcellular localization or half-life of protein 33 was observed. A possible role for protein 33 and protein 21 in the mechanism of action of luteinizing hormone on testosterone production of Leydig cells is discussed.