ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)
Biotechnologia Acta Т. 17, No. 2 , 2024
P. 56-56, Bibliography 2, Engl.
UDC::577.112; 577.217.535
DOI:https://doi.org/10.15407/biotech17.02.056
VALYL-TRNA SYNTHETASE INTERACTS WITH Β-SUBUNIT OF THE EUKARYOTIC TRANSLATION ELONGATION FACTOR COMPLEX eEF1B
N.T. Kolodka, V.F. Shalak, B.S. Negrutskii
Institute of Molecular Biology and Geneticsof the Natonal Academy of Scienses of Ukraine, Kyiv
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.
Kew words: valyl-tRNA synthetase, eukaryotic translation elongation factors, protein complexes, protein-protein interactions.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2024