Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone β-subunit cDNAs
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2017
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Plos One
Resumo
The 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.
Como referenciar
SEVILHANO, 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. Plos One, v. 12, n. 8, p. 1-22, 2017. DOI: 10.1371/journal.pone.0183545. Disponível em: http://repositorio.ipen.br/handle/123456789/28167. Acesso em: 29 Mar 2024.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.