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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone beta-subunit cDNAs

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Sevilhano, Thais ; de Carvalho, Roberto Feitosa ; de Jesus Oliveira, Nelio Alessandro ; Oliveira, Joao Ezequiel ; Maltarollo, Vinicius Goncalves ; Trossini, Gustavo ; Garcez, Riviane ; Bartolini, Paolo
Total Authors: 8
Document type: Journal article
Source: PLoS One; v. 12, n. 8 AUG 28 2017.
Web of Science Citations: 3

The common gonadotrophic hormone alpha-subunit (GTH alpha) has been previously isolated by our research group from A. gigas pituitaries; in the present work the cDNA sequences encoding FSH beta and LH beta subunits have also been isolated from the same species of fish. The FSH beta-subunit consists of 126 amino acids with a putative 18 amino acid signal peptide and a 108 amino acid mature peptide, while the LH beta-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 FSH beta and of Cypriniformes (76%) for LH beta, followed by Siluriformes, 53% for FSH beta and 75% for LH beta. Interestingly, the identity with the corresponding human amino acid sequences was still remarkable: 45.1% for FSH beta and 51.4% for LH beta. 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 ``seat-belt{''}, favored by a disulfide bond formed between the 3rd and 12th cysteine in both beta-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. (AU)

FAPESP's process: 12/10779-2 - Cloning, expression and characterization of Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) of pirarucu (Arapaima gigas)
Grantee:Paolo Bartolini
Support Opportunities: Regular Research Grants