Fluorogenic substrates for high-throughput measurements of endothelial lipase activity.
J Lipid Res
; 48(2): 472-82, 2007 Feb.
Article
em En
| MEDLINE
| ID: mdl-17090660
ABSTRACT
Endothelial lipase (EL) has been shown to be a critical determinant for high density lipoprotein cholesterol levels in vivo; therefore, assays that measure EL activity have become important for the discovery of small molecule inhibitors that specifically target EL. Here, we describe fluorescent Bodipy-labeled substrates that can be used in homogeneous, ultra-high-throughput kinetic assays that measure EL phospholipase or triglyceride lipase activities. Triton X-100 detergent micelles and synthetic HDL particles containing Bodipy-labeled phospholipid or Bodipy-labeled triglyceride substrates were shown to be catalytic substrates for EL, LPL, and HL. More importantly, only synthetic HDL particles containing Bodipy-labeled triglyceride were ideal substrates for EL, LPL, and HL in the presence of high concentrations of human or mouse serum. These data suggest that substrate presentation is a critical factor when determining EL activity in the presence of serum.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Corantes Fluorescentes
/
Lipase
Limite:
Animals
/
Humans
Idioma:
En
Revista:
J Lipid Res
Ano de publicação:
2007
Tipo de documento:
Article
País de afiliação:
Estados Unidos