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Registro Completo |
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Biblioteca(s): |
Embrapa Pesca e Aquicultura. |
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Data corrente: |
01/03/2023 |
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Data da última atualização: |
01/03/2023 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
FREIRE, R. P.; HERNANDEZ-GONZALEZ, J. E.; LIMA, E. R.; SUZUKI, M. F.; OLIVEIRA, J. E. de; TORATI, L. S.; BARTOLINI, P.; SOARES, C. R. J. |
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Afiliação: |
RENAN PASSOS FREIRE, IPEN-CNEN; JORGE ENRIQUE HERNANDEZ-GONZALEZ, UNESP-São Paulo, SP; ELIANA ROSA LIMA, IPEN-CNEN; MIRIAM FUSSAE SUZUKI, IPEN-CNEN; JOÃO EZEQUIEL DE OLIVEIRA, IPEN-CNEN; LUCAS SIMON TORATI, CNPASA; PAOLO BARTOLINI, IPEN-CNEN; CARLOS ROBERTO JORGE SOARES, IPEN-CNEN. |
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Título: |
Molecular cloning and AlphaFold modeling of Thyrotropin (ag-TSH) from the amazonian fish pirarucu (Arapaima gigas). |
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Ano de publicação: |
2023 |
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Fonte/Imprenta: |
Bioinformatics and Biology Insights, v. 17, p. 1-13, 2023. |
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ISSN: |
1177-9322 |
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DOI: |
https://doi.org/10.1177/11779322231154148 |
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Idioma: |
Inglês |
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Conteúdo: |
Arapaima gigas, known as Pirarucu in Brazil, is one of the largest freshwater fish in the world. Some individuals could reach 3m in length and weight up to 200kg. Due to extinction risks and its economic value, the species has been a focus for preservation and reproduction studies. Thyrotropin (TSH) is a glycoprotein hormone formed by 2 subunits a and b whose main activity is related to the synthesis of thyroid hormones (THs)?T3 and T4. In this work, we present a combination of bioinformatics tools to identify Arapaima gigas bTSH (ag-bTSH), modeling its molecular structure and express the recombinant heterodimer form in mammalian cells. Using the combination of computational biology, based on genome-related information, in silico molecular cloning and modeling led to confirm results of the ag-bTSH sequence by reverse transcriptase-polymerase chain reaction (RT-PCR) and transient expression in human embryonic kidney (HEK293F) cells. Molecular cloning of ag-bTSH retrieved 146 amino acids with a signal peptide of 21 amino acid residues and 6 disulfide bonds. The sequence has a similarity to 39 fish species, ranging between 43.1% and 81.6%, whose domains are extremely conserved, such as cystine knot motif and N-glycosylation site. The Arapaima gigas thyrotropin (ag-TSH) model, solved by AlphaFold, was used in molecular dynamics simulations with Scleropages formosus receptor, providing similar values of free energy AGbind and AGPMF in comparison with Homo sapiens model. The recombinant expression in HEK293F cells reached a yield of 25mg/L, characterized via chromatographic and physical-chemical techniques. This work shows that other Arapaima gigas proteins could be studied in a similar way, using the combination of these techniques, recovering more information from its genome and improving the reproduction and preservation of this prehistoric fish. MenosArapaima gigas, known as Pirarucu in Brazil, is one of the largest freshwater fish in the world. Some individuals could reach 3m in length and weight up to 200kg. Due to extinction risks and its economic value, the species has been a focus for preservation and reproduction studies. Thyrotropin (TSH) is a glycoprotein hormone formed by 2 subunits a and b whose main activity is related to the synthesis of thyroid hormones (THs)?T3 and T4. In this work, we present a combination of bioinformatics tools to identify Arapaima gigas bTSH (ag-bTSH), modeling its molecular structure and express the recombinant heterodimer form in mammalian cells. Using the combination of computational biology, based on genome-related information, in silico molecular cloning and modeling led to confirm results of the ag-bTSH sequence by reverse transcriptase-polymerase chain reaction (RT-PCR) and transient expression in human embryonic kidney (HEK293F) cells. Molecular cloning of ag-bTSH retrieved 146 amino acids with a signal peptide of 21 amino acid residues and 6 disulfide bonds. The sequence has a similarity to 39 fish species, ranging between 43.1% and 81.6%, whose domains are extremely conserved, such as cystine knot motif and N-glycosylation site. The Arapaima gigas thyrotropin (ag-TSH) model, solved by AlphaFold, was used in molecular dynamics simulations with Scleropages formosus receptor, providing similar values of free energy AGbind and AGPMF in comparison with Homo sapiens model. The recom... Mostrar Tudo |
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Thesagro: |
Peixe; Pirarucu. |
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Thesaurus Nal: |
Arapaima gigas; Fish; Molecular cloning; Prediction; Thyrotropin. |
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Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/doc/1152025/1/bbi-2023.pdf
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Marc: |
LEADER 02800naa a2200313 a 4500
001 2152025
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008 2023 bl uuuu u00u1 u #d
022 $a1177-9322
024 7 $ahttps://doi.org/10.1177/11779322231154148$2DOI
100 1 $aFREIRE, R. P.
245 $aMolecular cloning and AlphaFold modeling of Thyrotropin (ag-TSH) from the amazonian fish pirarucu (Arapaima gigas).$h[electronic resource]
260 $c2023
520 $aArapaima gigas, known as Pirarucu in Brazil, is one of the largest freshwater fish in the world. Some individuals could reach 3m in length and weight up to 200kg. Due to extinction risks and its economic value, the species has been a focus for preservation and reproduction studies. Thyrotropin (TSH) is a glycoprotein hormone formed by 2 subunits a and b whose main activity is related to the synthesis of thyroid hormones (THs)?T3 and T4. In this work, we present a combination of bioinformatics tools to identify Arapaima gigas bTSH (ag-bTSH), modeling its molecular structure and express the recombinant heterodimer form in mammalian cells. Using the combination of computational biology, based on genome-related information, in silico molecular cloning and modeling led to confirm results of the ag-bTSH sequence by reverse transcriptase-polymerase chain reaction (RT-PCR) and transient expression in human embryonic kidney (HEK293F) cells. Molecular cloning of ag-bTSH retrieved 146 amino acids with a signal peptide of 21 amino acid residues and 6 disulfide bonds. The sequence has a similarity to 39 fish species, ranging between 43.1% and 81.6%, whose domains are extremely conserved, such as cystine knot motif and N-glycosylation site. The Arapaima gigas thyrotropin (ag-TSH) model, solved by AlphaFold, was used in molecular dynamics simulations with Scleropages formosus receptor, providing similar values of free energy AGbind and AGPMF in comparison with Homo sapiens model. The recombinant expression in HEK293F cells reached a yield of 25mg/L, characterized via chromatographic and physical-chemical techniques. This work shows that other Arapaima gigas proteins could be studied in a similar way, using the combination of these techniques, recovering more information from its genome and improving the reproduction and preservation of this prehistoric fish.
650 $aArapaima gigas
650 $aFish
650 $aMolecular cloning
650 $aPrediction
650 $aThyrotropin
650 $aPeixe
650 $aPirarucu
700 1 $aHERNANDEZ-GONZALEZ, J. E.
700 1 $aLIMA, E. R.
700 1 $aSUZUKI, M. F.
700 1 $aOLIVEIRA, J. E. de
700 1 $aTORATI, L. S.
700 1 $aBARTOLINI, P.
700 1 $aSOARES, C. R. J.
773 $tBioinformatics and Biology Insights$gv. 17, p. 1-13, 2023.
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Registro original: |
Embrapa Pesca e Aquicultura (CNPASA) |
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| 1. |  | FREIRE, R. P.; HERNANDEZ-GONZALEZ, J. E.; LIMA, E. R.; SUZUKI, M. F.; OLIVEIRA, J. E. de; TORATI, L. S.; BARTOLINI, P.; SOARES, C. R. J. Molecular cloning and AlphaFold modeling of Thyrotropin (ag-TSH) from the amazonian fish pirarucu (Arapaima gigas). Bioinformatics and Biology Insights, v. 17, p. 1-13, 2023.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 2 |
| Biblioteca(s): Embrapa Pesca e Aquicultura. |
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