Busca avançada
Ano de início
Entree
(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Dansylglycine, a fluorescent probe for specific determination of halogenating activity of myeloperoxidase and eosinophil peroxidase

Texto completo
Autor(es):
Bertozo, Luiza de Carvalho ; Zeraik, Maria Luiza ; Ximenes, Valdecir Farias
Número total de Autores: 3
Tipo de documento: Artigo Científico
Fonte: Analytical Biochemistry; v. 532, p. 29-37, SEP 1 2017.
Citações Web of Science: 2
Resumo

Myeloperoxidase (MPO) and eosinophil peroxidase (EPO) are enzymes present in neutrophil and eosinophil leukocytes, respectively. Here, we present the development of a sensitive and specific assay for determination of the halogenating enzymatic activity of MPO and EPO based on the electrophilic attack of HOCI and HOBr on aromatic ring of dansylglycine (DG). We found that the intrinsic fluorescence of DG was promptly depleted by the action of these acids. In the presence of the enzymes, the fluorescence bleaching was dependent of chloride (Cl-) and bromide (Br-), which makes the assay able to distinguish the halogenating from the peroxidase activity. A linear correlation was obtained between the hydrogen peroxide (H2O2) concentration and the fluorescent decay. Similarly, the enzyme activity was measured by keeping constant H2O2. The method was applied for studding MPO/EPO specific inhibitors as 5-fluortryptamine (reversible inhibitor) and 4-hydroxybenzhydrazide (irreversible inhibitor). Differently of the taurine chloramine/3,3',5,5'-tetramethylbenzidine assay, which is among the most used technique, the dansylglycine assay was able to differentiate these inhibitors based on their kinetic behavior. In conclusion, this assay can differentiate the peroxidase and halogenating activity of MPO and EPO. Moreover, the method is adequate for real-time measurement of the production of HOCI and HOBr. (C) 2017 Elsevier Inc. All rights reserved. (AU)

Processo FAPESP: 15/21693-0 - Ácido gálico e seus ésteres como agentes anti -Helicobacter pylori e sequestradores de oxidantes produzidos por neutrófilos
Beneficiário:Vanessa Gonçalves Wolf
Linha de fomento: Bolsas no Brasil - Mestrado
Processo FAPESP: 16/20549-5 - Desenvolvimento e aplicação de sondas fluorescentes e baseadas em dicroísmo circular para estudos de interação de ligantes com proteínas, caracterização de proteínas amiloides e determinação de atividade enzimática
Beneficiário:Valdecir Farias Ximenes
Linha de fomento: Auxílio à Pesquisa - Regular