Berman A E, Gornaeva N P, Oborotova T A, Mazyrov V I
Biokhimiia. 1980 Jan;45(1):63-74.
The total protein-synthesizing and collagen-synthesizing activities of poly(A)-containing RNA from the membrane-bound polyribosomes of chicken embryos were studied under various conditions in a cell-free wheat germ system. The optimal concentrations of poly(A)-RNA, mono- and bivalent cations and spermine necessary for the translation of collagen and non-collagen proteins were determined. It was shown that within a wide range of K+ and Mg2+ concentrations in a cell-free system the poly(A)-RNAs studied are very actively involved in the translation of non-collagen proteins. At the same time the bacterial collagenase-cleavable polypeptides synthesized on poly(A)-RNA from the membrane-bound polyribosomes were identified only after addition of polyamine -- spermine (40 mk/M) to the system. In this case the relative level of collagen polypeptide synthesis remains constant within a wide range of concentrations of mRNA, K+ and Mg2+ in the system. Collagen-synthesizing activity of poly(A)-RNA from membrane-bound polyribosomes in a heterologous cell-free system as compared to that of membrane-bound polyribosomes in a homologous system showed that these polyribosomes can be effectively used for isolation of collagen mRNA.
在无细胞小麦胚芽系统的各种条件下，研究了鸡胚膜结合多核糖体中含聚腺苷酸（poly(A)）的RNA的总蛋白质合成和胶原蛋白合成活性。确定了翻译胶原蛋白和非胶原蛋白所需的poly(A)-RNA、单价和二价阳离子以及精胺的最佳浓度。结果表明，在无细胞系统中广泛的K+和Mg2+浓度范围内，所研究的poly(A)-RNA非常积极地参与非胶原蛋白的翻译。同时，只有在向系统中添加多胺——精胺（40μM）后，才能鉴定出在膜结合多核糖体的poly(A)-RNA上合成的可被细菌胶原酶切割的多肽。在这种情况下，在系统中mRNA、K+和Mg2+的广泛浓度范围内，胶原蛋白多肽合成的相对水平保持恒定。与同源系统中膜结合多核糖体相比，异源无细胞系统中膜结合多核糖体的poly(A)-RNA的胶原蛋白合成活性表明，这些多核糖体可有效地用于分离胶原蛋白mRNA。
