Hirai M, Asahi T
J Biochem. 1975 Sep;78(3):485-92. doi: 10.1093/oxfordjournals.jbchem.a130932.
Two RNases in bound forms associated with the microsomal membrane and with the ribosomes or unknown particles in pea root tissue were solubilized by subjecting the membrane to sonic oscillation in the presence of EDTA and KC1 and by treating the particles with EDTA, respectively. The RNases were than purified by DEAE-cellulose and Sephadex G-75 column chromatographies. The elution profiles of RNases from the columns were very similar. No significant differences were observed in their electrophoretic mobilities in polyacrylamide gels, in molecular weight, in activation by inorganic ions, urea or phospholipid micelles or in the dependence of their activities upon pH. The purified RNASES were not different from the bound enzymes as regards activation by inorganic ions and urea and the dependence of the activity upon pH. Triton X-100 stimulated the activity only if RNase was in a bound form associated with the microsomal membrane. We propose that the two RNases may be the same molecular species and differ only in the form of association with intracellular structures.
通过分别在存在乙二胺四乙酸（EDTA）和氯化钾（KC1）的情况下对膜进行超声振荡以及用EDTA处理颗粒，使豌豆根组织中与微粒体膜、核糖体或未知颗粒结合的两种核糖核酸酶（RNases）溶解。然后通过二乙氨基乙基纤维素（DEAE - 纤维素）和葡聚糖G - 75柱色谱法对核糖核酸酶进行纯化。核糖核酸酶从柱上的洗脱图谱非常相似。在聚丙烯酰胺凝胶中的电泳迁移率、分子量、无机离子、尿素或磷脂微胶粒的激活作用或其活性对pH的依赖性方面，均未观察到显著差异。就无机离子和尿素的激活作用以及活性对pH的依赖性而言，纯化后的核糖核酸酶与结合酶并无不同。只有当核糖核酸酶以与微粒体膜结合的形式存在时，聚山梨醇酯X - 100（Triton X - 100）才会刺激其活性。我们认为这两种核糖核酸酶可能是相同的分子种类，只是在与细胞内结构的结合形式上有所不同。
