A time-resolved fluorescence resonance energy transfer-based assay for DEN1 peptidase activity

被引：14

作者：

Engels, Ingo H. ^{[1
]}

Daguia, Catherine ^{[1
]}

Huynh, Truc ^{[1
]}

Urbina, Hugo ^{[1
]}

Buddenkotte, Joerg ^{[1
]}

Schumacher, Andrew ^{[1
]}

Caldwell, Jeremy S. ^{[1
]}

Brinker, Achim ^{[1
]}

机构：

[1] Genom Inst Novartis Res Fdn, San Diego, CA 92121 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2009年 / 390卷 / 01期

关键词：

UBIQUITIN-LIKE PROTEIN; NEDD8; ENZYMES; FAMILY; NEDP1; CUL1;

D O I：

10.1016/j.ab.2009.03.035

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

Neural precursor cell expressed, developmentally clown-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. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：85 / 87

页数：3

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