Isolation and Characterization of the Arapaima gigas Growth Hormone (ag-GH) cDNA and Three-Dimensional Modeling of This Hormone in Comparison with the Human Hormone (hGH)

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Lima, Eliana Rosa ; Freire, Renan Passos ; Suzuki, Miriam Fussae ; Oliveira, Joao Ezequiel ; Yosidaki, Vanessa Luna ; Peroni, Cibele Nunes ; Sevilhano, Thais ; Zorzeto, Moises ; Torati, Lucas Simon ; Soares, Carlos Roberto Jorge ; Lima, Igor Daniel de Miranda ; Kronenberger, Thales ; Maltarollo, Vinicius Goncalves ; Bartolini, Paolo

• Tipo de documento: Artigo Científico
• Fonte: BIOMOLECULES; v. 13, n. 1, p. 17-pg., 2023-01-01.

Resumo

In a previous work, the common gonadotrophic hormone alpha-subunit (ag-GTH alpha), the ag-FSH beta- and ag-LH beta-subunit cDNAs, were isolated and characterized by our research group from A. gigas pituitaries, while a preliminary synthesis of ag-FSH was also carried out in human embryonic kidney 293 (HEK293) cells. In the present work, the cDNA sequence encoding the ag-growth hormone (ag-GH) has also been isolated from the same giant Arapaimidae Amazonian fish. The ag-GH consists of 208 amino acids with a putative 23 amino acid signal peptide and a 185 amino acid mature peptide. The highest identity, based on the amino acid sequences, was found with the Elopiformes (82.0%), followed by Anguilliformes (79.7%) and Acipenseriformes (74.5%). The identity with the corresponding human GH (hGH) amino acid sequence is remarkable (44.8%), and the two disulfide bonds present in both sequences were perfectly conserved. Three-dimensional (3D) models of ag-GH, in comparison with hGH, were generated using the threading modeling method followed by molecular dynamics. Our simulations suggest that the two proteins have similar structural properties without major conformational changes under the simulated conditions, even though they are separated from each other by a >100 Myr evolutionary period (1 Myr = 1 million years). The sequence found will be used for the biotechnological synthesis of ag-GH while the ag-GH cDNA obtained will be utilized for preliminary Gene Therapy studies. (AU)

• Processo FAPESP: 20/16549-5 - Estudos preliminares relativos à aplicação da terapia gênica de hormônio de crescimento em Arapaima gigas
• Beneficiário: Paolo Bartolini
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 20/10435-8 - Síntese de gonadotrofina coriônica humana recombinante em células CHO e HEK-293, purificação e caracterização físico-química
• Beneficiário: Carlos Roberto Jorge Soares
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 17/26369-1 - Síntese, purificação e caracterização das gonadotrofinas de pirarucu (Arapaima gigas): ag-FSH e ag-LH
• Beneficiário: João Ezequiel de Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Regular