一种用于识别细胞中与特定RNA相连的氨基酸的新方法。

A new method for identifying the amino acid attached to a particular RNA in the cell.

作者信息

Suzuki T, Ueda T, Watanabe K

机构信息

Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, Japan.

出版信息

FEBS Lett. 1996 Mar 4;381(3):195-8. doi: 10.1016/0014-5793(96)00107-x.

DOI:10.1016/0014-5793(96)00107-x

PMID:8601454

Abstract

To investigate the function of tRNAs or any other aminoacylable RNAs in vivo, it is important to be able to estimate the amounts and species of aminoacylated RNAs in living cells. We have developed a method of analyzing amino acids attached to particular tRNAs obtained from cells. After the ester bond between the amino acid and the 3'-adenosine moiety of a specific aminoacyl-tRNA is stabilized by acetylation of the amino acid with [14C]acetic anhydride, the aminoacyl-tRNA can be fished out with a solid-phase-attached DNA probe. The 14C-labeled acetylamino acid is then released from the thus purified acetyl-aminoacyl-tRNAs by alkaline treatment and detected by TLC analysis.

摘要

为了研究体内转运RNA（tRNA）或任何其他可氨酰化RNA的功能，能够估计活细胞中氨酰化RNA的数量和种类非常重要。我们开发了一种分析从细胞中获得的特定tRNA所连接氨基酸的方法。在用[¹⁴C]乙酸酐使氨基酸乙酰化从而稳定氨基酸与特定氨酰-tRNA的3'-腺苷部分之间的酯键后，可以用固相连接的DNA探针钓出氨酰-tRNA。然后通过碱性处理从如此纯化的乙酰氨酰-tRNA中释放出¹⁴C标记的乙酰氨基酸，并通过薄层色谱分析进行检测。