SEVILHANO, THAISCARVALHO, ROBERTO F. deOLIVEIRA, NELIO A. de J.OLIVEIRA, JOAO E.MALTAROLLO, VINICIUS G.TROSSINI, GUSTAVOGARCEZ, RIVIANEBARTOLINI, PAOLO2017-12-182017-12-182017SEVILHANO, THAIS; CARVALHO, ROBERTO F. de; OLIVEIRA, NELIO A. de J.; OLIVEIRA, JOAO E.; MALTAROLLO, VINICIUS G.; TROSSINI, GUSTAVO; GARCEZ, RIVIANE; BARTOLINI, PAOLO. Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone β-subunit cDNAs. <b>Plos One</b>, v. 12, n. 8, p. 1-22, 2017. DOI: <a href="https://dx.doi.org/10.1371/journal.pone.0183545">10.1371/journal.pone.0183545</a>. Disponível em: http://repositorio.ipen.br/handle/123456789/28167.1932-6203http://repositorio.ipen.br/handle/123456789/28167The common gonadotrophic hormone α-subunit (GTHα) has been previously isolated by our research group from A. gigas pituitaries; in the present work the cDNA sequences encoding FSHβ and LHβ subunits have also been isolated from the same species of fish. The FSH β-subunit consists of 126 amino acids with a putative 18 amino acid signal peptide and a 108 amino acid mature peptide, while the LH β-subunit consists of 141 amino acids with a putative 24 amino acid amino acid signal peptide and a 117 amino acid mature peptide. The highest identity, based on the amino acid sequences, was found with the order of Anguilliformes (61%) for FSHβ and of Cypriniformes (76%) for LHβ, followed by Siluriformes, 53% for FSHβ and 75% for LHβ. Interestingly, the identity with the corresponding human amino acid sequences was still remarkable: 45.1% for FSHβ and 51.4% for LHβ. Three dimensional models of ag-FSH and ag-LH, generated by using the crystal structures of h-FSH and h-LH as the respective templates and carried out via comparative modeling and molecular dynamics simulations, suggested the presence of the so-called ªseat-beltº, favored by a disulfide bond formed between the 3rd and 12th cysteine in both β-subunits. The sequences found will be used for the biotechnological synthesis of A. gigas gonadotrophic hormones (ag-FSH and ag-LH). In a first approach, to ascertain that the cloned transcripts allow the expression of the heterodimeric hormones, ag-FSH has been synthesized in human embryonic kidney 293 (HEK293) cells, preliminarily purified and characterized.1-22openAccesshormonespituitary glandpituitary hormonesgonadotropinsfishesluteinizing hormonedna-cloningmolecular modelsArtigo de periódico81210.1371/journal.pone.018354577.34