Abstract
When different mutations were introduced into the anticodon loop and at position 73 of YFA2, a derivative of yeast tRNAPhe, a single tRNA body was misacylated with 13 different amino acids. The affinities of these misacylated tRNAs for Thermus thermophilus elongation factor Tu (EF-Tu)·GTP were determined using a ribonuclease protection assay. A range of 2.5 kcal/mol in the binding energies was observed, clearly demonstrating that EF-Tu specifically recognizes the side chain of the esterified amino acid. Furthermore, this specificity can be altered by introducing a mutation in the amino acid binding pocket on the surface of EF-Tu. Also, when discussed in conjunction with the previously determined specificity of EF-Tu for the tRNA body, these experiments further demonstrate that EF-Tu uses thermodynamic compensation to bind cognate aminoacyl-tRNAs similarly.
Original language | English (US) |
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Pages (from-to) | 6159-6166 |
Number of pages | 8 |
Journal | Biochemistry |
Volume | 43 |
Issue number | 20 |
DOIs | |
State | Published - May 25 2004 |
ASJC Scopus subject areas
- Biochemistry