Nakamura M, Watanuki M, Hall P F
Mol Cell Endocrinol. 1978 Nov;12(2):209-19. doi: 10.1016/0303-7207(78)90115-6.
Y-1 adrenal tumor cells were incubated with aminoglutethimide with and without ACTH. Greater production of pregnenolone from endogenous cholesterol was observed (after washing to remove aminoglutethimide) in mitochondria from cells incubated with aminoglutethimide and ACTH than in those from cells incubated with aminoglutethimide alone. This response was inhibited by cycloheximide and puromycin but not by chloramphenicol or actinomycin D. ACTH increased the incorporation of [3H]tyrosine into protein associated with mitochondria but not into total cell protein or protein of postmitochondrial supernatant. This response did not require aminoglutethemide block and was inhibited by cycloheximide and puromycin but not by chloramphenicol or actinomycin D. Dibutyryl cyclic AMP produced both of these responses (increased production of pregnenolone and synthesis of protein associated with mitochondria). The concentration of cycloheximide required to cause 50% inhibition of the responses to ACTH and dibutyryl cyclic AMP was approximately the same for steroidogenesis by whole cells, for production of pregnenolone by isolated mitochondria, for incorporation of [3H]tyrosine into Y-1 cell protein and for the increase in synthesis of protein associated with mitochondria produced by ACTH (0.08--0.2 microgram/ml). Disc gel electrophoresis revealed that the increased incorporation of [3H]tyrosine involved two proteins corresponding to molecular weight of approximately 27,000 and 13,000 respectively. These observations suggest that ACTH promotes synthesis of protein(s) by cytoplasmic ribosomes on stable messenger RNA, that the protein(s) becomes associated with mitochondria and that the protein(s) includes one or more which are associated with the increase in production of pregnenolone produced in mitochondria by the addition of ACTH to adrenal cells.
将Y-1肾上腺肿瘤细胞分别在有和没有促肾上腺皮质激素（ACTH）的情况下与氨鲁米特一起孵育。在洗涤以去除氨鲁米特后，观察到与单独用氨鲁米特孵育的细胞相比，用氨鲁米特和ACTH孵育的细胞的线粒体中，内源性胆固醇生成孕烯醇酮的量更多。这种反应受到环己酰亚胺和嘌呤霉素的抑制，但不受氯霉素或放线菌素D的抑制。ACTH增加了[3H]酪氨酸掺入与线粒体相关的蛋白质中，但没有增加掺入总细胞蛋白质或线粒体后上清液中的蛋白质中。这种反应不需要氨鲁米特阻断，并且受到环己酰亚胺和嘌呤霉素的抑制，但不受氯霉素或放线菌素D的抑制。二丁酰环磷腺苷产生了这两种反应（增加孕烯醇酮的生成和与线粒体相关的蛋白质的合成）。对于全细胞的类固醇生成、分离线粒体中孕烯醇酮的生成、[3H]酪氨酸掺入Y-1细胞蛋白质以及ACTH引起的与线粒体相关的蛋白质合成增加，导致对ACTH和二丁酰环磷腺苷反应50%抑制所需的环己酰亚胺浓度大致相同（0.08 - 0.2微克/毫升）。圆盘凝胶电泳显示，[3H]酪氨酸掺入增加涉及两种蛋白质，其分子量分别约为27,000和13,000。这些观察结果表明，ACTH通过稳定的信使核糖核酸促进细胞质核糖体合成蛋白质，该蛋白质与线粒体结合，并且该蛋白质包括一种或多种与向肾上腺细胞添加ACTH后线粒体中孕烯醇酮生成增加相关的蛋白质。
