Biopolym. Cell. 2008; 24(1):21-27.
Structure and Function of Biopolymers
Isolation and purification of Thermus thermophilus tRNALys and determination of its modified nucleotides
1Krikliviy I. A., 1Kovalenko O. P., 1Gudzera O. Y., 1Yaremchuk A. D., 1Tukalo M. A.
- Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
Abstract
Lysyl-tRNA synthetase along with aspartyl- and asparagyl-tRNA synthetase belong to subclass IIb and have a number of specific features. On the one hand, subclass IIb 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 tRNALys and the determination of modified bases in its structure.
Keywords: тРНКLys, chromatography, extremal thermophyle Thermus thermophilus, benzoylated diethylaminoethylcellulose, modified nucleotides
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