Curgy J J, Ledoigt G, Stevens B J, André J
J Cell Biol. 1974 Mar;60(3):628-40. doi: 10.1083/jcb.60.3.628.
Mitochondrial and cytoplasmic ribosomes from Tetrahymena pyriformis have been isolated and studied by the techniques of polyacrylamide gel electrophoresis and electron microscopy used in conjunction. Although the two ribosome types show the same coefficient of sedimentation (80S) in sucrose gradients, they can be distinguished by gel electrophoresis: mitoribosomes migrate in a single band, considerably slower than the cytoribosome band. Electron microscope observations of negatively stained cytoribosomes show typical rounded or triangular profiles, about 275 x 230 A; mitoribosome profiles are much larger and clearly elongate, about 370 x 240 A. An electron-opaque spot delimits two nearly equal size subunits. In mixtures of mito- and cytoribosomes, each type can be recognized by its characteristic electrophoretic mobility and by its distinctive fine structure. Cytoribosomal 60S and 40S subunits each produce a distinct electrophoretic band. On the contrary, neither electrophoretic analysis, using a variety of conditions, nor electron microscopy is able to discern two different subunit types in the single 55S mitoribosomal subunit peak. Electrophoretic analysis of RNA shows that both ribosomal RNA species are present in the mitoribosomal subunit fraction. These results establish that mitoribosomes from T. pyriformis dissociate into two subunits endowed with the same sedimentation coefficient, the same electrophoretic mobility, and a similar morphology.
梨形四膜虫的线粒体核糖体和细胞质核糖体已通过聚丙烯酰胺凝胶电泳技术和电子显微镜技术相结合的方法进行了分离和研究。尽管这两种核糖体类型在蔗糖梯度中显示出相同的沉降系数(80S),但它们可以通过凝胶电泳加以区分:线粒体核糖体迁移成单一一条带,比细胞质核糖体带迁移得慢得多。对经负染的细胞质核糖体进行电子显微镜观察,显示出典型的圆形或三角形轮廓,约275×230埃;线粒体核糖体轮廓要大得多且明显呈细长形,约370×240埃。一个电子不透明斑点界定了两个大小几乎相等的亚基。在线粒体核糖体和细胞质核糖体的混合物中,每种类型都可通过其特征性的电泳迁移率和独特的精细结构来识别。细胞质核糖体的60S和40S亚基各自产生一条独特的电泳带。相反,无论采用何种条件进行电泳分析,还是电子显微镜观察,都无法在单一的55S线粒体核糖体亚基峰中辨别出两种不同的亚基类型。对RNA的电泳分析表明,两种核糖体RNA种类都存在于线粒体核糖体亚基组分中。这些结果表明,梨形四膜虫的线粒体核糖体可解离成两个具有相同沉降系数、相同电泳迁移率和相似形态的亚基。