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

Engels, Ingo and Daguia, Catherine and Huynh, Truc and Urbina, Hugo and Buddenkotte, Joerg and Schumacher, Andrew and Caldwell, Jeremy and Brinker, Achim (2009) A time-resolved fluorescence resonance energy transfer-based assay for DEN1 peptidase activity. Analytical Biochemistry, 390 (1). pp. 85-87. ISSN 1096-0309

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.

Item Type: Article
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Additional Information: author can archive post-print (ie final draft post-refereeing); Publisher's version/PDF cannot be used
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Date Deposited: 14 Dec 2009 13:49
Last Modified: 31 Jan 2013 00:57
URI: https://oak.novartis.com/id/eprint/1211

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