SIEKEVITZ P, PALADE G E
J Biophys Biochem Cytol. 1960 Jul;7(4):631-44. doi: 10.1083/jcb.7.4.631.
Ribonucleoprotein (RNP)(1) particles isolated by DOC treatment from pancreatic microsomes have a RNA content of 35 to 45 per cent of their dry weight. In the analytical ultracentrifuge about 85 per cent of the material has a sedimentation coefficient of approximately 85 S. These particles contain amylase, RNase, and trypsin-activatable proteolytic activities which cannot be washed off or detached by incubation in 0.44 M sucrose. The enzymes are released, however, by incubation in the presence of low concentrations of ATP, PP, or EDTA, and high concentrations of IP and AMP. At the same time, and at the same concentrations, approximately 80 per cent of the RNA and approximately 25 per cent of the protein of the particles becomes also non-sedimentable. The simultaneous addition of Mg(++) to the incubation medium prevents these losses. This finding, together with the observation that all the Mg(++) of the particles is released by the same agents, makes it likely that Mg(++) holds the particles together, and that its removal by the chelators used causes the particles to disintegrate. These findings are discussed in relation to the molecular structure of the RNP particles.
通过用去氧胆酸钠(DOC)处理从胰腺微粒体中分离得到的核糖核蛋白(RNP)(1)颗粒,其RNA含量占干重的35%至45%。在分析超速离心机中,约85%的物质沉降系数约为85S。这些颗粒含有淀粉酶、核糖核酸酶以及可被胰蛋白酶激活的蛋白水解活性,在0.44M蔗糖中孵育无法将这些活性洗脱或去除。然而,在低浓度的ATP、焦磷酸(PP)或乙二胺四乙酸(EDTA)以及高浓度的肌醇磷酸(IP)和腺苷一磷酸(AMP)存在下孵育,酶会被释放出来。与此同时,在相同浓度下,颗粒中约80%的RNA和约25%的蛋白质也变得不可沉降。向孵育介质中同时添加镁离子(Mg(++))可防止这些损失。这一发现,连同颗粒中所有的镁离子都被相同试剂释放这一观察结果,表明镁离子使颗粒聚集在一起,而所用螯合剂将其去除会导致颗粒解体。结合RNP颗粒的分子结构对这些发现进行了讨论。