Biopolym. Cell. 1993; 9(6):31-35.
Structure and Function of Biopolymers
Conformational change of tyrosyl-tRNA synthetase from bovine liver in the course of cognate tRNA binding revealed from fluorescence spectroscopy data
1Klimenko I. V., 1Kornelyuk A. I., 1Matsuka G. Kh.
- Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
Abstract
The fluorescent probe 1,5-1-AEDANS was covalently attached to tyrosyl-tRNA synthetase from bovine liver in a nearly stoichiometric amount (2 molecules of probe per enzyme dimer). Singlet-singlet energy transfer has been used for measurement of the apparent distance, between 6 tryptophan residues of enzyme and covalently attached 1,5-1-AEDANS. This distance was about 27.4 A assuming the random orientation of the donor and acceptor from polarization measurements. The interaction of cognate. tRNATyr with bovine tyrosyl-tRNA synthetase also resulted in the enhancement of fluorescence of AEDANS-labelled tyrosyRRNA synthetase (25 %) due to the increase of energy transfer efficiency. After binding of cognate bovine tRNATyr the apparent distance between tryptophan residues and AEDANS probe reduced to 25.3 A E. coli tRNATyr which was not aminoacylated by bovine enzyme did not induced these effects. Adding of ATP to the complex of bovine tRNATyr and tyrosyl-tRNA synthetase exhibited further enhancement of AEDANS fluorescence by 32 %. In this case the apparent distance between tryptophan residues and AEDANS probe was 24.1 A. These results are consistent with the conformational change of mammalian tyrosyl-tRNA synthetase, in the course of recognition of cognate tRNATyr.
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