ROBERTO FEITOSA DE CARVALHO
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Artigo IPEN-doc 23455 Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone β-subunit cDNAs2017 - SEVILHANO, THAIS; CARVALHO, ROBERTO F. de; OLIVEIRA, NELIO A. de J.; OLIVEIRA, JOAO E.; MALTAROLLO, VINICIUS G.; TROSSINI, GUSTAVO; GARCEZ, RIVIANE; BARTOLINI, PAOLOThe 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.Resumo IPEN-doc 21356 Molecular cloning, characterization and phylogenetic analysis of pirarucu (Arapaima gigas) FSH and LH beta-subunits2015 - BARTOLINI, PAOLO; CARVALHO, ROBERTO F.; SEVILHANO, THAIS C. dos A.; OLIVEIRA, JOAO E.; GARCEZ, RIVIANEResumo IPEN-doc 18704 Glicoprotein alpha characterization of the Amazonia fis (Arapaima gigas)2012 - CARVALHO, R.F.; SEVILHANO, T.C.A.; OLIVEIRA, N.A.J.; SOARES, C.R.J.; OLIVEIRA, J.E.; BARTOLINI, P.; SANTOS, P.R.E.; FARIA M.T.Artigo IPEN-doc 18812 Isolation of the pituitary gonadotrophic 'alpha'-subunit hormone of the giant amazonian fish: pirarucu (Arapaima gigas)2013 - FARIA, M.T.; CARVALHO, R.F.; SEVILHANO, T.C.A.; OLIVEIRA, N.A.J.; SILVA, C.F.P.; OLIVEIRA, J.E.; SOARES, C.R.J.; GARCEZ, R.; SANTO, P.R.E.; BARTOLINI, P.