VALYL-TRNA SYNTHETASE INTERACTS WITH Β-SUBUNIT OF THE EUKARYOTIC TRANSLATION ELONGATION FACTOR COMPLEX eEF1B
Journal: Biotechnologia Acta (Vol.17, No. 2)Publication Date: 2024-04-30
Authors : N.T. Kolodka V.F. Shalak B.S. Negrutskii;
Page : 56-56
Keywords : valyl-tRNA synthetase; eukaryotic translation elongation factors; protein complexes; protein-protein interactions;
Abstract
The aim of this study was to investigate the interaction of the N-terminal domain of the valyl-tRNA synthetase with α, β, and γ subunits of the eEF1B translation elongation factor complex. Methods: for this purpose, all 4 proteins were synthesized in bacterial cells and purified to homogeneity by a combination of chromatographic methods. The interaction of the eEF1B complex subunits with the N-terminal domain of the valyl-tRNA synthetase was verified by gel filtration and in vitro pull-down assays. Protein fractions collected at these stages were analyzed by SDS-PAGE. Results: according to the gel filtration results, eEF1Bα and eEF1Bγ subunits do not form a stable complex with the valine-tRNA synthetase domain. The potential for complexation of the eEF1Bβ subunit was evaluated by pull-down assay, which showed that this protein does interact with the valyl-tRNA synthetase. Conclusions: we concluded that the eEF1Bα and eEF1Bγ subunits do not interact with the valyl-tRNA synthetase compared to the eEF1Bβ protein. The N-terminal domain of the valyl-tRNA synthetase is necessary and sufficient for this interaction.
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