Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone β-subunit cDNAs
| Autor: | Riviane Garcez, João E. Oliveira, Vinícius Gonçalves Maltarollo, Roberto Feitosa de Carvalho, Nélio A. J. Oliveira, T. C. A. Sevilhano, Paolo Bartolini, Gustavo Henrique Goulart Trossini |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Glycosylation Glycobiology lcsh:Medicine Peptide Molecular Dynamics Biochemistry Database and Informatics Methods Computational Chemistry 0302 clinical medicine Cloning Molecular Amino Acids Post-Translational Modification lcsh:Science chemistry.chemical_classification Multidisciplinary Organic Compounds Fishes Complementary DNA Amino acid Nucleic acids Chemistry Pituitary Gland Follicle Stimulating Hormone beta Subunit Physical Sciences PIRARUCU Sequence Analysis Research Article Signal peptide DNA Complementary Proline Forms of DNA Bioinformatics Fish Biology 030209 endocrinology & metabolism Sequence alignment Molecular cloning Research and Analysis Methods 03 medical and health sciences Genetics Fish Physiology Animals Humans Sulfur Containing Amino Acids Animal Physiology Cysteine Biology and life sciences lcsh:R Organic Chemistry Chemical Compounds Proteins Cyclic Amino Acids Luteinizing Hormone beta Subunit DNA Hormones Vertebrate Physiology HEK293 Cells 030104 developmental biology chemistry lcsh:Q Sequence Alignment Zoology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP PLoS ONE PLoS ONE, Vol 12, Iss 8, p e0183545 (2017) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0183545 |
| Popis: | 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 "seat-belt", 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. |
| Databáze: | OpenAIRE |
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