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Insights into the structure, function and stability of bordonein-L, the first L-amino acid oxidase from Crotalus durissus terrificus snake venom.
Wiezel, Gisele A; Rustiguel, Joane K; Morgenstern, David; Zoccal, Karina F; Faccioli, Lucia H; Nonato, M Cristina; Ueberheide, Beatrix; Arantes, Eliane C.
Afiliação
  • Wiezel GA; Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil.
  • Rustiguel JK; Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil.
  • Morgenstern D; Proteomics Resource Center, Langone Medical Center, New York University, 430 East 29th St, 8th Floor, 10016, New York, NY, USA.
  • Zoccal KF; Department of Clinical Analysis, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil.
  • Faccioli LH; Department of Clinical Analysis, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil.
  • Nonato MC; Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil.
  • Ueberheide B; Proteomics Resource Center, Langone Medical Center, New York University, 430 East 29th St, 8th Floor, 10016, New York, NY, USA.
  • Arantes EC; Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, SP, Brazil. Electronic address: ecabraga@fcfrp.usp.br.
Biochimie ; 163: 33-49, 2019 Aug.
Article em En | MEDLINE | ID: mdl-31078582
Snake venom L-amino acid oxidases (svLAAOs) are an interesting class of enzymes with important biological activities. Their participation in key metabolic processes, including pathological disorders, suggest that svLAAOs are potential lead compounds in drug discovery. However, their short-term stability defies their applications. This paper describes the stability studies together with functional and structural characterization of the LAAO bordonein-L. It has 498 amino acid residues, one N-glycosylation site and two disulfide bonds, revealed by high-resolution MS/MS. Molecular modeling approach showed its monomer folds into three conserved domains: FAD, substrate and helical domains. Differential scanning fluorimetry showed the enzyme tends to destabilize from neutral to basic pHs and in presence of mono/bivalent ions and it is highly stabilized by acid pHs and its substrates. However, high concentrations of L-amino acids decrease bordonein-L enzyme activity. Dynamic light scattering revealed bordonein-L remains in the dimeric and monodisperse form, so aggregation does not cause the rapidly decrease of enzyme activity. In vitro, the enzyme exhibited cytotoxicity against fibroblast cell line and killed Leishmania amazonensis promastigotes, intensified by substrate addition. Concluding, our results provide biochemistry and biophysical insights to improve LAAOs stability and better approaches to long-term storage. Moreover, our study emphasizes the importance of proper buffers choice mainly in cell-based assays.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Venenos de Serpentes / Crotalus / L-Aminoácido Oxidase Limite: Animals Idioma: En Revista: Biochimie Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: França

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Venenos de Serpentes / Crotalus / L-Aminoácido Oxidase Limite: Animals Idioma: En Revista: Biochimie Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: França