Biopolym. Cell. 2003; 19(3):231-237.
http://dx.doi.org/10.7124/bc.000654
Structure and Function of Biopolymers
Isolation and characterization of monoclonal antibodies against eukaryotic translation elongation factor 1A
1Pogrebnaya A. P., 2Markeyeva N. V., 3Lisogubov V. V., 4Fridberg A. M., 4Zabashnyi S. I., 3Usenko V. S., 1Filonenko V. V., 2Pogrebnoy P. V., 1Negrutskii B. S.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. R. E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine
    45, Vasilkivska Str., Kyiv, Ukraine, 01022
  3. Pathomorphological Laboratory "BIONTEK"
    52A/88, Komsomol'ska Str., Dnipropetrovs'k, Ukraine, 49000
  4. Dnipropetrovsk Regional Clinical Hospital Mechnikov
    Sq. October, 14, Dnepropetrovsk, Ukraine, 49008

Abstract

Eukaryotic translation elongation factor 1A (eEF1A) is one of the main components of the translation apparatus.The protein also fulfills a number of non-canonical functions not related directly to protein synthesis. In this work the homogenous elongation factor 1A was obtained from hen liver by several chromatographic procedures with different carriers. The purified protein was used to develop monoclonal antibodies. The anti-eEF1A antibodies obtained are species-unspecific, they recognize both eEF1A and eEF1A2 isoforms in ELJSA and immunoblotting. The antibodies have been successfully used for immunohistological studies on human tissues.
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References

[1] Negrutskii BS, El’skaya AV. Eukaryotic Translation Elongation Factor 1?: Structure, Expression, Functions, and Possible Role in Aminoacyl-tRNA Channeling. Progress in Nucleic Acid Research and Molecular Biology. 1998;47–78.
[2] Lee S, Wolfraim LA, Wang E. Differential expression of S1 and elongation factor-1 alpha during rat development. J Biol Chem. 1993;268(32):24453-9.
[3] Lee S, LeBlanc A, Duttaroy A, Wang E. Terminal differentiation-dependent alteration in the expression of translation elongation factor-1 alpha and its sister gene, S1, in neurons. Exp Cell Res. 1995;219(2):589-97.
[4] Knudsen SM, Frydenberg J, Clark BF, Leffers H. Tissue-dependent variation in the expression of elongation factor-1 alpha isoforms: isolation and characterisation of a cDNA encoding a novel variant of human elongation-factor 1 alpha. Eur J Biochem. 1993;215(3):549-54.
[5] Cavallius J, Rattan SI, Clark BF. Changes in activity and amount of active elongation factor 1 alpha in aging and immortal human fibroblast cultures. Exp Gerontol. 1986;21(3):149-57.
[6] Talapatra S, Wagner JD, Thompson CB. Elongation factor-1 alpha is a selective regulator of growth factor withdrawal and ER stress-induced apoptosis. Cell Death Differ. 2002;9(8):856-61.
[7] Ohta K, Toriyama M, Miyazaki M, Murofushi H, Hosoda S, Endo S, Sakai H. The mitotic apparatus-associated 51-kDa protein from sea urchin eggs is a GTP-binding protein and is immunologically related to yeast polypeptide elongation factor 1 alpha. J Biol Chem. 1990;265(6):3240-7.
[8] Tatsuka M, Katayama H, Ota T, Tanaka T, Odashima S, Suzuki F, Terada Y. Multinuclearity and increased ploidy caused by overexpression of the aurora- and Ipl1-like midbody-associated protein mitotic kinase in human cancer cells. Cancer Res. 1998;58(21):4811-6.
[9] Wu-Baer F, Lane WS, Gaynor RB. Identification of a group of cellular cofactors that stimulate the binding of RNA polymerase II and TRP-185 to human immunodeficiency virus 1 TAR RNA. J Biol Chem. 1996;271(8):4201-8.
[10] Durso NA, Cyr RJ. A calmodulin-sensitive interaction between microtubules and a higher plant homolog of elongation factor-1 alpha. Plant Cell. 1994;6(6):893-905.
[11] Yang W, Boss WF. Regulation of phosphatidylinositol 4-kinase by the protein activator PIK-A49. Activation requires phosphorylation of PIK-A49. J Biol Chem. 1994;269(5):3852-7.
[12] Grant AG, Flomen RM, Tizard ML, Grant DA. Differential screening of a human pancreatic adenocarcinoma lambda gt11 expression library has identified increased transcription of elongation factor EF-1 alpha in tumour cells. Int J Cancer. 1992;50(5):740-5.
[13] Anand N, Murthy S, Amann G, Wernick M, Porter LA, Cukier IH, Collins C, Gray JW, Diebold J, Demetrick DJ, Lee JM. Protein elongation factor EEF1A2 is a putative oncogene in ovarian cancer. Nat Genet. 2002;31(3):301-5.
[14] Su Z, Goldstein NI, Fisher PB. Antisense inhibition of the PTI-1 oncogene reverses cancer phenotypes. Proc Natl Acad Sci U S A. 1998;95(4):1764-9.
[15] Kaziro Y. The role of guanosine 5'-triphosphate in polypeptide chain elongation. Biochim Biophys Acta. 1978;505(1):95-127.
[16] Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.
[17] Kristensen P, Lund A, Clark BF, Cavallius J, Merrick WC. Purification and characterisation of a tissue specific elongation factor 1 alpha (EF-1 alpha 2) from rabbit muscle. Biochem Biophys Res Commun. 1998;245(3):810-4.
[18] Kinzy TG, Merrick WC. Characterization of a limited trypsin digestion form of eukaryotic elongation factor 1 alpha. J Biol Chem. 1991;266(7):4099-105.
[19] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[20] Filonenko VV, Wolfson AD, Wartanyan OA, Beresten SF. Monoclonal antibodies to the components of the high-molecular-mass aminoacyl-tRNA synthetase complex. Biomed Sci. 1991;2(3):289-92.
[21] Lorette C. J. Immunocytochemical methods and protocols. Ed. C. J. Lorette. Tokyo: Humana press, 1999:233 p.
[22] 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.
[23] Davydova EK, Sitikov AS, Ovchinnikov LP. Phosphorylation of elongation factor 1 in polyribosome fraction of rabbit reticulocytes. FEBS Lett. 1984;176(2):401-5.
[24] Dever TE, Costello CE, Owens CL, Rosenberry TL, Merrick WC. Location of seven post-translational modifications in rabbit elongation factor 1 alpha including dimethyllysine, trimethyllysine, and glycerylphosphorylethanolamine. J Biol Chem. 1989;264(34):20518-25.
[25] Tisdale EJ, Tartakoff AM. Extensive labeling with [3H]ethanolamine of a hydrophilic protein of animal cells. J Biol Chem. 1988;263(17):8244-52.
[26] Venema RC, Peters HI, Traugh JA. Phosphorylation of elongation factor 1 (EF-1) and valyl-tRNA synthetase by protein kinase C and stimulation of EF-1 activity. J Biol Chem. 1991;266(19):12574-80.