Wiezel, Gisele A.
Rustiguel, Joane K.
Zoccal, Karina E.
Faccioli, Lucia H.
Cristina Nonato, M.
Arantes, Eliane C.
Número total de Autores: 8
Afiliação do(s) autor(es):
 Univ Sao Paulo, Sch Pharmaceut Sci Ribeirao Preto, Dept Phys & Chem, Av Cafe S-N, BR-14040903 Ribeirao Preto, SP - Brazil
 NYU, Prote Resource Ctr, Langone Med Ctr, 430 East 29th St, 8th Floor, New York, NY 10016 - USA
 Weizmann Inst Sci, De Botton Inst Prot Profiling, Nancy & Stephen Grand Israel Natl Ctr Personalize, IL-76100 Rehovot - Israel
 Univ Sao Paulo, Dept Clin Anal Toxicol & Food Sci, Sch Pharmaceut Sci Ribeirao Preto, Av Cafe S-N, BR-14040903 Ribeirao Preto, SP - Brazil
 Ctr Univ Barao Maua, Rua Ramos de Azevedo 423, BR-14090180 Ribeirao Preto, SP - Brazil
Número total de Afiliações: 5
Tipo de documento:
Citações Web of Science:
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. (C) 2019 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved. (AU)