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A new method for identifying the amino acid attached to a particular RNA in the cell
Author(s) -
Suzuki Tsutomu,
Ueda Takuya,
Watanabe Kimitsuna
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00107-x
Subject(s) - rna , chemistry , computational biology , cell , amino acid , biochemistry , microbiology and biotechnology , biology , gene
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 [ 14 C]acetic anhydride, the aminoacyl‐tRNA can be fished out with a solid‐phase‐attached DNA probe. The 14 C‐labeled acetylamino acid is then released from the thus purified acetyl‐aminoacyl‐tRNAs by alkaline treatment and detected by TLC analysis.