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)

Author(s):Show less -

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

• Document type: Journal article
• Source: BIOMOLECULES; v. 13, n. 1, p. 17-pg., 2023-01-01.

Abstract

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)

• FAPESP's process: 20/16549-5 - preliminary studies directed to the application of growth hormone gene therapy to Arapaima gigas
• Grantee: Paolo Bartolini
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/10435-8 - Synthesis of recombinant human chorionic gonadotrophine in CHO and HEK-293 cells, purification and physicochemical characterization
• Grantee: Carlos Roberto Jorge Soares
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/26369-1 - Synthesis, purification and characterization of the gonadotropins of pirarucu (Arapaima gigas): ag-FSH and ag-LH
• Grantee: João Ezequiel de Oliveira
• Support Opportunities: Regular Research Grants