Molecular architecture of leishmania EF-1α reveals a novel site that may modulate protein translation: A possible target for drug development

Abstract: Elongation factor-1α plays an essential role in eukaryotic protein biosynthesis. Recently, we have shown by protein structure modeling the presence of a hairpin-loop of 12 amino acids in mammalian EF-1α that is absent in the leishmania homologue [D. Nandan, A. Cherkasov, R. Sabouti, T. Yi, N.E. Reiner, Molecular cloning, biochemical and structural analysis of elongation factor-1 alpha from Leishmania donovani: comparison with the mammalian homologue, Biochem. Biophys. Res. Commun. 302 (2003) 646–652]. As a consequence of this deletion, an exposed region is available on the main body of leishmania EF-1α. Here we report the generation of an anti-EF-1α antibody (DN-3) which bound selectively to the exposed region of leishmania EF-1α, with no reactivity with human EF-1α. In a leishmania cell-free protein translation system, DN-3 substantially inhibited protein translation. A similar inhibitory effect was observed when a specific peptide based on the exposed region was used in the cell-free protein translation assay. The application of structure-based in silico methods to identify potential ligands to target the exposed region identified a small molecule that selectively attenuated in vitro translation using leishmania extracts. Moreover, this small molecule showed selective suppressive effect on multiplication of leishmania in culture. Taken together, these findings identify a novel, exposed region in leishmania EF-1α that may be involved in protein synthesis and a potential site for drug targeting. [Copyright &y& Elsevier]