Bakalkin G I, Kal'nov S L, Zubatov A S, Luzikov V N
Biokhimiia. 1976 Oct;41(10):1878-88.
Estimation of the rate of degradation of the products of mitochondrial protein synthesis in S. cerevisiae cells is reported. The method developed for this purpose is based on pulse incorporation of a labeled amino acid in the presence of an inhibitor of cytoplasmic protein synthesis and allows one to monitor postincorporation of the label. The label incorporated is shown to be rapidly released from mitochondria. Its content is decreased 2-fold during 20-30 min at the beginning and 50-60 min at the end of the exponential phase of growth. The label is detected in cytosol proteins and the TCA-soluble fraction of mitochondria, which is indicative of possible proteolysis of mitochondrial membrane proteins. Since release of the label does not undergo inhibition by specific inhibitors of yeast cell proteinases (pepstatin and phenylmethylsulfonyl fluoride), it may be assumed that these proteinases are either not involved in the digestion of the products of mitochondrial protein synthesis or do not represent a rate-limiting step of the process.
本文报道了酿酒酵母细胞中线粒体蛋白质合成产物降解速率的估计。为此目的开发的方法基于在细胞质蛋白质合成抑制剂存在下标记氨基酸的脉冲掺入,并允许监测标记掺入后的情况。结果表明,掺入的标记物会迅速从线粒体中释放出来。在生长指数期开始时,其含量在20 - 30分钟内减少2倍,在指数期末期50 - 60分钟内减少2倍。在细胞质蛋白和线粒体的三氯乙酸可溶性部分中检测到该标记物,这表明线粒体膜蛋白可能发生了蛋白水解。由于标记物的释放不受酵母细胞蛋白酶的特异性抑制剂(胃蛋白酶抑制剂和苯甲基磺酰氟)的抑制,因此可以推测这些蛋白酶要么不参与线粒体蛋白质合成产物的消化,要么不代表该过程的限速步骤。