A time-resolved fluorescence resonance energy transfer-based assay for DEN1 peptidase activity.
Anal Biochem
; 390(1): 85-7, 2009 Jul 01.
Article
em En
| MEDLINE
| ID: mdl-19328766
ABSTRACT
Neural precursor cell expressed, developmentally down-regulated gene 8 (NEDD8) is a recently discovered ubiquitin-like posttranslational modifier. NEDD8 acts predominantly as a regulator of ubiquitin-protein ligases and as a decoy for proteins targeted for proteasomal degradation. It thereby controls key events in cell cycle progression and embryogenesis. Deneddylase-1 (DEN1/NEDP1/SENP8) features a selective peptidase activity converting the proNEDD8 precursor to its mature form and an isopeptidase activity deconjugating NEDD8 from substrates such as cullins and p53. In this study, we describe a high-throughput screening (HTS)-compatible time-resolved fluorescent resonance energy transfer (TR-FRET) assay measuring the peptidase activity of DEN1.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Endopeptidases
/
Transferência Ressonante de Energia de Fluorescência
Limite:
Humans
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
2009
Tipo de documento:
Article