Chen C S, Wen T N, Tuan H M
Biochim Biophys Acta. 1982 Nov 30;699(2):92-7. doi: 10.1016/0167-4781(82)90142-7.
It has been previously shown that Clostridium sticklandii specifically synthesized three readily separable 75Se-labeled tRNAs, designated seleno-tRNAs I, II and III, and the partially purified seleno-tRNA II cochromatographed with L-prolyl-tRNA on DEAE-Sephadex A-50 (Chen, C.S. and Stadtman, T.C. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 1403-1407). In the present study a highly purified 75Se-labeled tRNA I was obtained by chromatography on benzoylated DEAE-cellulose, DEAE-Sephadex A-50 and Sepharose 4B. The 75Se-labeled tRNA I cochromatographed with an L-valine-accepting species on DEAE-Sephadex A-50 and Sepharose 4B. Addition of a 285-fold molar excess of unlabeled L-valine to the L-valine acceptor activity assay mixture markedly decreased the amount of L-[14C]valine bound to seleno-tRNA I.
先前已经表明，史氏梭菌能特异性合成三种易于分离的75Se标记的tRNA，分别命名为硒代tRNA I、II和III，部分纯化的硒代tRNA II在DEAE-葡聚糖A-50上与L-脯氨酰-tRNA共层析（陈，C.S.和斯塔特曼，T.C.（1980年）《美国国家科学院院刊》77，1403 - 1407）。在本研究中，通过在苯甲酰化DEAE-纤维素、DEAE-葡聚糖A-50和琼脂糖4B上进行层析，获得了高度纯化的75Se标记的tRNA I。75Se标记的tRNA I在DEAE-葡聚糖A-50和琼脂糖4B上与一种L-缬氨酸接受型物质共层析。在L-缬氨酸受体活性测定混合物中加入285倍摩尔过量的未标记L-缬氨酸，显著降低了与硒代tRNA I结合的L-[14C]缬氨酸的量。
