Jackson M, Ingle J
Biochem J. 1973 Mar;131(3):523-33. doi: 10.1042/bj1310523.
The aggregation of ribosomal RNA species during chromatography on methylated albumin on kieselguhr was decreased from 50 to 15% by the lower salt concentrations made possible by the use of higher pH values. The polydisperse RNA was resolved into two fractions. About 50% was eluted with the rRNA whereas the remainder was bound to the column, and was recovered only by solubilization of the methylated albumin. Both fractions of polydisperse RNA were similar in size range, but the bound fraction was considerably richer in AMP. No D-RNA (DNA-like RNA) peak was resolved under these conditions of column fractionation. However, the properties of the bound RNA were consistent with it containing both D-RNA and TB-RNA (tenaciously bound RNA). The relationship between these two fractions of AMP-rich RNA was considered. The bound RNA and ribosomal RNA responded differently to various treatments. The salt concentration required to elute ribosomal RNA was halved by increasing the pH of the fractionation, but the amount of bound RNA was in fact increased. Denaturation by hot urea decreased the binding of ribosomal RNA to the methylated albumin, but did not facilitate elution of bound RNA. The high affinity between the AMP-rich polydisperse RNA and the methylated albumin does not therefore appear to arise for the secondary structure conferred by the high AMP content.
在硅藻土上的甲基化白蛋白柱层析过程中,通过使用较高pH值使盐浓度降低,核糖体RNA种类的聚集从50%降至15%。多分散RNA被分离成两个组分。约50%与核糖体RNA一起被洗脱,而其余部分则结合在柱上,只有通过溶解甲基化白蛋白才能回收。多分散RNA的两个组分在大小范围内相似,但结合组分中的AMP含量要丰富得多。在这些柱分级分离条件下,未分辨出D-RNA(类DNA RNA)峰。然而,结合RNA的性质与它同时含有D-RNA和TB-RNA(紧密结合RNA)一致。考虑了富含AMP的RNA的这两个组分之间的关系。结合RNA和核糖体RNA对各种处理的反应不同。通过提高分级分离的pH值,洗脱核糖体RNA所需的盐浓度减半,但结合RNA的量实际上增加了。热尿素变性降低了核糖体RNA与甲基化白蛋白的结合,但并未促进结合RNA的洗脱。因此,富含AMP的多分散RNA与甲基化白蛋白之间的高亲和力似乎并非由高AMP含量赋予的二级结构引起。