Myoglobin microplate assay to evaluate prevention of protein peroxidation

• Tipo: Artigo em Revista Científica Internacional
• Data: 2015

Autores

Marques, SS (Autor) Outra; Magalhaes, LM (Autor) Outra; Mota, AIP (Autor) Outra; Soares, TRP (Autor) Outra; Korsak, B (Autor) Outra; Reis, S (Autor) Outra; Segundo, MA (Autor) FFUP

Revista

Título: Journal of Pharmaceutical and Biomedical Analysis

Vol. 114

Páginas: 305-311

ISSN: 0731-7085

Editora: Elsevier

Indexação

ISI Web of Knowledge - 1 Citação

Scopus - 2 Citações

Outras Informações

ID Authenticus: P-00G-CH6

DOI: 10.1016/j.jpba.2015.06.006

Abstract (EN): The current therapeutic strategies are based on the design of multifunctional drug candidates able to interact with various disease related targets. Drugs that have the ability to scavenge reactive oxygen species (ROS), beyond their main therapeutic action, may prevent the oxidative damage of biomolecules. Therefore, analytical approaches that monitor in a continuous mode the ability of drugs to counteract peroxidation of physiologically relevant biotargets are required. In the present work, a microplate spectrophotometric assay is proposed to evaluate the ability of selected cardiovascular drugs, including angiotensin-converting enzyme (ACE) inhibitors, beta-blockers and statins to prevent protein peroxidation. Myoglobin, which is a heme protein, and peroxyl radicals generated from thermolysis of 2,2'-azo-bis(2-amidinopropane) dihydrochloride at 37 C, pH 7.4 were selected as protein model and oxidative species, respectively. Myoglobin peroxidation was continuously monitored by the absorbance decrease at 409 nm and the ability of drugs to counteract protein oxidation was determined by the calculation of the area under the curve upon the myoglobin oxidation. Fluvastatin (AUC(50) = 12.5 +/- 1.2 mu M) and enalapril (AUC(50) = 15.2 +/- 1.8 mu M) showed high ability to prevent myoglobin peroxidation, providing even better efficiency than endogenous antioxidants such as reduced glutathione. Moreover, labetalol, enalapril and fluvastatin prevent the autoxidation of myoglobin, while glutathione showed a pro-oxidant effect.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 7