Registration Number
0225U004153,
(0120U102238)
, R & D reports
Title
Structural and functional studies of translation elongation factors of higher eukaryotes
popup.stage_title
Вивчення просторової будови субодиниць eEF1Bα, eEF1Bβ i eEF1Bγ комплексу факторів трансляції eEF1B із використанням воднево-дейтерієвого обміну із подальшою мас-спектрометрією та інших фізико-хімічних методів.
Head
Negrutskyii Borys S., Доктор біологічних наук
Registration Date
20-11-2025
Organization
Institute of Molecular Biology and Genetics of NAS of Ukraine
popup.description1
The aim of this study is to analyze the spatial organization and function of mammalian translation elongation factor 1 (eEF1A and the multi-protein eEF1B complex). To achieve this, the following tasks are planned. First, to obtain experimental data that will allow us to build atomic models of the spatial organization of the subunits of the eEF1B translation complex and the architecture of the complex as a whole. Second, to conduct experimental studies and to build the model of spatial organization of isoform 1 of elongation factor 1A based on them. Third, to determine the potential functional significance of methylation of lysine residues in the molecule of elongation factor 1A (eEF1A), in particular, to test the hypothesis experimentally that the methylation of some lysine residues in eEF1A may affect the interaction of this protein with partners , such as the subunits of the eEF1B complex, which may reveal the molecular mechanism of the recently discovered effect of eEF1A methylation on oncogenic cell transformation. Fourthly, describe the molecular mechanisms of action of novel eEF1A inhibitors with potential antitumor effect.
popup.description2
Objects of study – the human translation factor complex eEF1B, the translation elongation factor eEF1A, its isoforms eEF1A1 and eEF1A2, and the elongation factors eEF1Bα, eEF1Bβ, and eEF1Bγ.
Objective of the work – to elucidate the spatial organization of translation complexes and their components, and to investigate the potential role of post-translational modifications in the functioning of translation factors.
Research methods – hydrogen–deuterium exchange followed by mass spectrometry, analytical ultracentrifugation, protein mutagenesis, chromatographic purification of individual proteins, analytical gel filtration, bioluminescence resonance energy transfer (BRET) system, Western blotting, co-immunoprecipitation, protein co-precipitation, confocal microscopy, mass spectrometry, polymerase chain reaction (PCR), molecular biology methods for DNA manipulation, kinetic methods, bioinformatic analysis, molecular modeling, and molecular docking.
Measurements of the hydrodynamic radius of eEF1Bα were performed using the dynamic light scattering method with adaptive correlation. The obtained value was 3.35 nm. This criterion was used to select the optimal three-dimensional structural model of this protein.
Using X-ray scattering, the molecular shape of eEF1Bβ was assessed, which was essential for selecting the optimal three-dimensional structural model of this protein.
Analytical ultracentrifugation showed that eEF1Bγ, depending on concentration, readily forms homo-associates of varying multimeric states. The eEF1Bγ monomer is a moderately elongated, non-globular protein in solution.
Product Description
popup.authors
Novosylna Oleksandra V.
Hanna V. Yelska
Porublova Larysa V.
Вячеслав Ф. Шалак
Ihor L. Lysetskiy
Ganna Lukyanenko
popup.nrat_date
2025-11-20
