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Updated: Aug 19, 2026

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
Published on: February 25, 2011
Conformational change of mammalian tyrosyl-tRNA synthetase induced by tyrosyl adenylate formation
A I Kornelyuk1, I V Klimenko, K A Odynets
1Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kiev.
Abstract:
The fluorescent probe 1,5-I-AEDANS was covalently attached to bovine tyrosyl-tRNA synthetase outside of enzyme active site in a nearly stoichiometric amount (2 probe molecules per enzyme dimer). Singlet-singlet resonance energy transfer has been used for the measurement of the apparent distance between tryptophan residues of enzyme and covalently attached 1,5-I-AEDANS. This distance was estimated as 27.4 A in the assumption of the random orientation of the donor and acceptor fluorophores. Tyrosyl adenylate formation catalyzed by bovine tyrosyl-tRNA synthetase resulted in the highly specific enhancement of 1,5-I-AEDANS fluorescence and concomitant decrease of the apparent distance between the probe and tryptophanyls to 22.3-25.7 A. These results are consistent with the conformational change of tyrosyl-tRNA synthetase during tyrosyl adenylate formation which propagates to distant from active site regions of enzyme structure.
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