Simpkins H, Panko E, Tay S
Biochem J. 1973 Oct;135(2):299-305. doi: 10.1042/bj1350299.
Because it has been proposed that the ribosome-membrane interaction is different in endoplasmic reticulum derived from a non-secretory and secretory cell we undertook a study to determine whether attachment of the ribosome to the membrane involved ribosomal RNA and if the rRNA in ribosomes derived from the two classes of cell possessed an altered susceptibility to RNAase (ribonuclease) hydrolysis. We found that brain ribosomes appeared to possess more regions accessible to nuclease attack, independent of whether a sequence-dependent RNAase (T(1)) or a sterically hindered RNAase bound to Enzite polymer was employed. These results were independent of whether the ribosomes were membrane-bound or detached from the endoplasmic reticulum membranes, but at high RNAase concentration these differences became negligible. No conclusions, however, could be drawn as to whether ribosomal RNA is involved in the attachment of the ribosome to the endoplasmic reticulum membrane, because of the presence of endogeneous membrane-associated RNAases. Analysis of the rRNA fragments by polyacrylamide-gel electrophoresis suggests that the sites available for attack by low concentrations of nuclease in bound-ribosomes derived from brain cortex are different from those of liver.
由于有人提出源自非分泌细胞和分泌细胞的内质网中核糖体与膜的相互作用不同,我们开展了一项研究,以确定核糖体与膜的附着是否涉及核糖体RNA,以及源自这两类细胞的核糖体中的rRNA对RNA酶(核糖核酸酶)水解的敏感性是否有所改变。我们发现,无论使用序列依赖性RNA酶(T(1))还是与Enzite聚合物结合的空间位阻RNA酶,脑核糖体似乎都有更多易受核酸酶攻击的区域。这些结果与核糖体是结合在膜上还是从内质网膜上分离无关,但在高RNA酶浓度下,这些差异变得微不足道。然而,由于存在内源性膜相关RNA酶,因此无法就核糖体RNA是否参与核糖体与内质网膜的附着得出结论。通过聚丙烯酰胺凝胶电泳对rRNA片段进行分析表明,源自大脑皮层的结合核糖体中低浓度核酸酶可攻击的位点与肝脏中的不同。