Liu D K, Matrisian P E
Biochem J. 1977 May 15;164(2):371-7. doi: 10.1042/bj1640371.
Kidney cortex, red medulla and white medulla were separated into nuclei, mitochondria, microsomal and 105000g supernatant fractions. Assay of RNAase (ribonuclease) activity at pH7.8 revealed that, for each subcellular fraction, activity was much greater in cortex than in red or white medulla; this was true for both free RNAase and total (free plus latent) RNAase. For example, the free RNAase activity in the 105000g supernatant of cortex was 5 and 8 times higher than in red and white medulla respectively. No latent RNAase activity was found in any particulate fraction. Latent supernatant RNAase activities (suggesting presence of bound RNAase inhibitor) were similar in cortex and medulla. The cortex supernatant contained minimal free RNAase inhibitor, whereas that of the red and white medulla showed about one-third and one-tenth respectively of the inhibitor activity measured in liver. Adrenalectomy did not change RNAase activity in any fraction nor the content of free RNAase inhibitor in the kidney supernatant, but did decrease the liver RNAase inhibitor content by 40%. In supernatants from mouse kidney, both free and total RNAase activities of both cortex and red medulla were similar to those of rat red medulla. Mouse cortex contained appreciably higher amounts of free RNAase inhibitor than rat cortex. The difference between the rat and mouse cortical RNAase activity and inhibitor content may help explain the relative ease with which satisfactory renal polyribosome profiles were obtained from mouse kidneys. Our results, as well as those of Kline & Liberti [(1973) Biochem. Biophys. Res. Commun.52, 1271-1277], showing that renal red and white medulla are more active than cortex in protein synthesis, are consistent with the hypothesis that the RNAase-RNAase-inhibitor system may participate in the regulation of protein synthesis.
肾皮质、肾红髓和肾白髓被分离成细胞核、线粒体、微粒体以及105000g上清液组分。在pH7.8条件下对核糖核酸酶(RNAase)活性进行测定,结果显示,对于每个亚细胞组分,皮质中的活性均远高于红髓或白髓;游离RNAase和总(游离加潜在)RNAase均是如此。例如,皮质105000g上清液中的游离RNAase活性分别比红髓和白髓高5倍和8倍。在任何微粒体组分中均未发现潜在RNAase活性。皮质和髓质中潜在的上清液RNAase活性(提示存在结合型RNAase抑制剂)相似。皮质上清液中游离RNAase抑制剂极少,而红髓和白髓的上清液中分别显示出肝脏中测得的抑制剂活性的约三分之一和十分之一。肾上腺切除术未改变任何组分中的RNAase活性,也未改变肾上清液中游离RNAase抑制剂的含量,但使肝脏RNAase抑制剂含量降低了40%。在小鼠肾的上清液中,皮质和红髓的游离和总RNAase活性均与大鼠红髓的相似。小鼠皮质中游离RNAase抑制剂的含量明显高于大鼠皮质。大鼠和小鼠皮质RNAase活性及抑制剂含量的差异可能有助于解释从小鼠肾中更容易获得令人满意的肾多核糖体图谱的原因。我们的结果以及Kline和Liberti [(1973年)《生物化学与生物物理学研究通讯》52, 1271 - 1277] 的结果表明,肾红髓和白髓在蛋白质合成方面比皮质更活跃,这与RNAase - RNAase抑制剂系统可能参与蛋白质合成调节的假说一致。