Study of different physical and chemical parameters of non-canonical complex of eukaryotic translation elongation factor 1A with deacylated tRNA
DOI:
https://doi.org/10.7124/bc.000572Abstract
For the first time the KA of [eEF-1A·GDP] with free tRNA was determined by steady state fluorescence polarization. The obtained value – 20 nM is allowed to suppose a physiological significance of the complex investigated. For the first time the solution conformation of rabbit liver [eEF-1A·GDP] and its complex with tRNA has been studied by small-angle neutron scattering (SANS). The shape of the [eEF-1A·GDP] molecule in solution is demonstrated to be rather relaxed and to undergo rather substantial changes upon formation of the complex with tRNA. The complex has a more compact structure than free [eEF-1A · GDP].References
Petrushenko ZM, Negrutskii BS, Ladokhin AS, Budkevich TV, Shalak VF, El'skaya AV. Evidence for the formation of an unusual ternary complex of rabbit liver EF-1alpha with GDP and deacylated tRNA. FEBS Lett. 1997;407(1):13-7.
Negrutskii BS, Deutscher MP. Channeling of aminoacyl-tRNA for protein synthesis in vivo. Proc Natl Acad Sci U S A. 1991;88(11):4991-5.
Shalak VF, Budkevich TV, Negrutski? BS, El'skaia AV. A fast and effective method for purification of elongation factor 1 alpha from rabbit liver. Ukr Biokhim Zh. 1997;69(2):104-9.
Brunngraber EF. A simplified procedure for the preparation of "soluble" RNA from rat liver. Biochem Biophys Res Commun. 1962;8:1-3.
Weiel J, Hershey JW. Fluorescence polarization studies of the interaction of Escherichia coli protein synthesis initiation factor 3 with 30S ribosomal subunits. Biochemistry. 1981;20(20):5859-65.
Jameson DM, Seifried SE. Quantification of protein-protein interactions using fluorescence polarization. Methods. 1999;19(2):222-33.
Graifer DM, Nekhai SYu, Mundus DA, Fedorova OS, Karpova GG. Interaction of human and Escherichia coli tRNA(Phe) with human 80S ribosomes in the presence of oligo- and polyuridylate templates. Biochim Biophys Acta. 1992;1171(1):56-64.
Bilgin N, Ehrenberg M, Ebel C, Zaccai G, Sayers Z, Koch MH, Svergun DI, Barberato C, Volkov V, Nissen P, Nyborg J. Solution structure of the ternary complex between aminoacyl-tRNA, elongation factor Tu, and guanosine triphosphate. Biochemistry. 1998;37(22):8163-72.
