Biopolymers and cell. 2008; 24 (1): 21 - 27

Isolation and purification of Thermus thermophilus tRNA^Lys and determination of its modified nucleotides

I. A. Krikliviy, O. P. Kovalenko, O. Y. Gudzera, A. D. Yaremchuk, M. A. Tukalo

Lysyl-tRNA synthetase along with aspartyl- and asparagyl-tRNA synthetase belong to subclass ²²b and have a number of specific features. On the one hand, subclass ²²b synthetases have N-terminal anticodon-linking domains (~140 residues), function as homodimers a2, and anticodon triplets for each ARSase are the main but insufficient identity elements. On the other hand, all tRNA anticodons, corresponding to each synthetase, contain a central U, and discrimination between them often depends on one nucleotide only. Thus, the questions are posed, concerning the structural basis and the mechanisms for discrimination between these anticodons by a homologous binding domain, as well as the possible role of modified bases in this respect. In this paper we described methodological approach to obtaining pure Thermus thermophilus tRNA^Lys and the determination of modified bases in its structure.

Keywords: tRNA^Lys, chromatography, extremal thermophyle Thermus thermophilus, benzoylated diethylaminoethylcellulose, modified nucleotides.