title: Biophysical and structural characterization of the recombinant human eIF3L.
abstract: The eukaryotic translation initiation factor 3, subunit L (eIF3L) is one of the subunits of the eIF3 complex, an accessory protein of the Polymerase I enzyme and may have an important role in the Flavivirus replication by interaction with a viral non-structural 5 protein. Considering the importance of eIF3L in a diversity of cellular functions, we have produced the recombinant full-length eIF3L protein in Escherichia coli and performed spectroscopic and in silico analyses to gain insights into its hydrodynamic behavior and structure. Dynamic light scattering showed that eIF3L behaves as monomer when it is not interacting with other molecular partners. Circular dichroism experiments showed a typical spectrum of α-helical protein for eIF3L, which is supported by sequence-based predictions of secondary structure and the 3D in silico model. The molecular docking with the K subunit of the eIF3 complex revealed a strong interaction. It was also predicted several potential interaction sites in eIF3L, indicating that the protein is likely capable of interacting with other molecules as experimentally shown in other functional studies. Moreover, bioinformatics analyses showed approximately 8 putative phosphorylation sites and one possible N-glycosylation site, suggesting its regulation by post-translational modifications. The production of the eIF3L protein in E. coli and structural information gained in this study can be instrumental for target-based drug design and inhibitors against Flavivirus replication and to shed light on the molecular mechanisms involved in the eukaryotic translation initiation.
authors.forename: Ana T S
authors.initials: AT
authors.lastname: Morais
authors.forename: Andreia N
authors.initials: AN
authors.lastname: Meza
authors.forename: Gabriela C
authors.initials: GC
authors.lastname: Araújo
authors.forename: Alessandra
authors.initials: A
authors.lastname: Vidotto
authors.forename: Fátima P
authors.initials: FP
authors.lastname: Souza
authors.forename: Marcelo A
authors.initials: MA
authors.lastname: Fossey
authors.forename: Mário T
authors.initials: MT
authors.lastname: Murakami
authors.affiliation: Laboratório de Pesquisa em Virologia, Faculdade de Medicina de Rio Preto, Avenida Brigadeiro Faria Lima 5416, Vila São Pedro, São José do Rio Preto, SP, 15090-000, Brazil. mnogueira@famerp.br.
authors.forename: Mauricio L
authors.initials: ML
authors.lastname: Nogueira
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D039621:Eukaryotic Initiation Factor-3 / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*; D006031:Glycosylation; D006801:Humans; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000235:genetics*
chemical_list: C443114:EIF3L protein, human; D039621:Eukaryotic Initiation Factor-3; D011994:Recombinant Proteins