Biopolym. Cell. 2000; 16(4):281-283.
http://dx.doi.org/10.7124/bc.000572
Structure and Function of Biopolymers
Study of different physical and chemical parameters of non-canonical complex of eukaryotic translation elongation factor 1A with deacylated tRNA
1Budkevich T. V., 1Negrutskii B. S., 2Serdyuk I. N., 1El'skaya A. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Institute of Protein Research, Russian Academy of Sciences
    Pushchino, Moscow Region, Russian Federation, 142290

Abstract

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].
Full text: (PDF, in Russian)

References

[1] 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.
[2] 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.
[3] 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.
[4] Brunngraber EF. A simplified procedure for the preparation of "soluble" RNA from rat liver. Biochem Biophys Res Commun. 1962;8:1-3.
[5] 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.
[6] Jameson DM, Seifried SE. Quantification of protein-protein interactions using fluorescence polarization. Methods. 1999;19(2):222-33.
[7] 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.
[8] 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.