Development of a high-throughput fluorescence polarization DNA cleavage assay for the identification of FEN1 inhibitors.
J Biomol Screen
; 18(5): 567-75, 2013 Jun.
Article
em En
| MEDLINE
| ID: mdl-23427045
ABSTRACT
Flap endonuclease-1 (FEN1) is a highly conserved metallonuclease and is the main human flap endonuclease involved in the recognition and cleavage of single-stranded 5' overhangs from DNA flap structures. The involvement of FEN1 in multiple DNA metabolism pathways and the identification of FEN1 overexpression in a variety of cancers has led to interest in FEN1 as an oncology target. In this article, we describe the development of a 1536-well high-throughput screening assay based on the change in fluorescence polarization of a FEN1 DNA substrate labeled with Atto495 dye. The assay was subsequently used to screen 850 000 compounds from the AstraZeneca compound collection, with a Z' factor of 0.66 ± 0.06. Hits were followed up by IC50 determination in both a concentration-response assay and a technology artifact assay.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Endonucleases Flap
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Inibidores Enzimáticos
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Clivagem do DNA
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Descoberta de Drogas
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Ensaios de Triagem em Larga Escala
Tipo de estudo:
Diagnostic_studies
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Prognostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article