A biosensor of S100A4 metastasis factor activation: inhibitor screening and cellular activation dynamics.
Biochemistry
; 47(3): 986-96, 2008 Jan 22.
Article
em En
| MEDLINE
| ID: mdl-18154362
ABSTRACT
S100A4, a member of the S100 family of Ca2+-binding proteins, displays elevated expression in malignant human tumors compared with benign tumors, and increased expression correlates strongly with poor patient survival. S100A4 has a direct role in metastatic progression, likely due to the modulation of actomyosin cytoskeletal dynamics, which results in increased cellular motility. We developed a fluorescent biosensor (Mero-S100A4) that reports on the Ca2+-bound, activated form of S100A4. Direct attachment of a novel solvatochromatic reporter dye to S100A4 results in a sensor that, upon activation, undergoes a 3-fold enhancement in fluorescence, thus providing a sensitive assay for use in vitro and in vivo. In cells, localized activation of S100A4 at the cell periphery is observed during random migration and following stimulation with lysophosphatidic acid, a known activator of cell motility and proliferation. Additionally, a screen against a library of FDA-approved drugs with the biosensor identified an array of phenothiazines as inhibitors of myosin-II associated S100A4 function. These data demonstrate the utility of the new biosensor both for drug discovery and for probing the cellular dynamics controlled by the S100A4 metastasis factor.
Texto completo:
1
Coleções:
01-internacional
Temas:
Geral
Base de dados:
MEDLINE
Assunto principal:
Técnicas Biossensoriais
/
Proteínas S100
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Cálcio
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Fibroblastos
Tipo de estudo:
Diagnostic_studies
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Prognostic_studies
/
Screening_studies
Idioma:
En
Revista:
Biochemistry
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Estados Unidos