<?xml version="1.0" encoding="UTF-8"?><xml><records><record><source-app name="Biblio" version="7.x">Drupal-Biblio</source-app><ref-type>17</ref-type><contributors><authors><author><style face="normal" font="default" size="100%">van der Linden, Lonneke</style></author><author><style face="normal" font="default" size="100%">Ulferts, Rachel</style></author><author><style face="normal" font="default" size="100%">Nabuurs, Sander B</style></author><author><style face="normal" font="default" size="100%">Kusov, Yuri</style></author><author><style face="normal" font="default" size="100%">Liu, Hong</style></author><author><style face="normal" font="default" size="100%">George, Shyla</style></author><author><style face="normal" font="default" size="100%">Lacroix, Céline</style></author><author><style face="normal" font="default" size="100%">Goris, Nesya</style></author><author><style face="normal" font="default" size="100%">David Lefebvre</style></author><author><style face="normal" font="default" size="100%">Lanke, Kjerstin H W</style></author><author><style face="normal" font="default" size="100%">Kris De Clercq</style></author><author><style face="normal" font="default" size="100%">Hilgenfeld, Rolf</style></author><author><style face="normal" font="default" size="100%">Neyts, Johan</style></author><author><style face="normal" font="default" size="100%">van Kuppeveld, Frank J M</style></author></authors></contributors><titles><title><style face="normal" font="default" size="100%">Application of a cell-based protease assay for testing inhibitors of picornavirus 3C proteases.</style></title><secondary-title><style face="normal" font="default" size="100%">Antiviral Res</style></secondary-title><alt-title><style face="normal" font="default" size="100%">Antiviral Res</style></alt-title></titles><keywords><keyword><style  face="normal" font="default" size="100%">3C Viral Proteases</style></keyword><keyword><style  face="normal" font="default" size="100%">Animals</style></keyword><keyword><style  face="normal" font="default" size="100%">Antiviral Agents</style></keyword><keyword><style  face="normal" font="default" size="100%">Cell Line</style></keyword><keyword><style  face="normal" font="default" size="100%">Cysteine Endopeptidases</style></keyword><keyword><style  face="normal" font="default" size="100%">Drug Evaluation, Preclinical</style></keyword><keyword><style  face="normal" font="default" size="100%">Genes, Reporter</style></keyword><keyword><style  face="normal" font="default" size="100%">Humans</style></keyword><keyword><style  face="normal" font="default" size="100%">Luciferases, Firefly</style></keyword><keyword><style  face="normal" font="default" size="100%">Picornaviridae</style></keyword><keyword><style  face="normal" font="default" size="100%">Protease Inhibitors</style></keyword><keyword><style  face="normal" font="default" size="100%">Viral Proteins</style></keyword></keywords><dates><year><style  face="normal" font="default" size="100%">2014</style></year><pub-dates><date><style  face="normal" font="default" size="100%">2014 Mar</style></date></pub-dates></dates><volume><style face="normal" font="default" size="100%">103</style></volume><pages><style face="normal" font="default" size="100%">17-24</style></pages><language><style face="normal" font="default" size="100%">eng</style></language><abstract><style face="normal" font="default" size="100%">&lt;p&gt;Proteolytical cleavage of the picornaviral polyprotein is essential for viral replication. Therefore, viral proteases are attractive targets for anti-viral therapy. Most assays available for testing proteolytical activity of proteases are performed in vitro, using heterologously expressed proteases and peptide substrates. To deal with the disadvantages associated with in vitro assays, we modified a cell-based protease assay for picornavirus proteases. The assay is based on the induction of expression of a firefly luciferase reporter by a chimeric transcription factor in which the viral protease and cleavage sites are inserted between the GAL4 binding domain and the VP16 activation domain. Firefly luciferase expression is dependent on cleavage of the transcription factor by the viral protease. This biosafe assay enables testing the effect of compounds on protease activity in cells while circumventing the need for infection. We designed the assay for 3C proteases (3C(pro)) of various enteroviruses as well as of viruses of several other picornavirus genera, and show that the assay is amenable for use in a high-throughput setting. Furthermore, we show that the spectrum of activity of 3C(pro) inhibitor AG7088 (rupintrivir) not only encompasses enterovirus 3C(pro) but also 3C(pro) of foot-and-mouth disease virus (FMDV), an aphthovirus. In contrary, AG7404 (compound 1), an analogue of AG7088, had no effect on FMDV 3C(pro) activity, for which we provide a structural explanation.&lt;/p&gt;</style></abstract><custom1><style face="normal" font="default" size="100%">https://www.ncbi.nlm.nih.gov/pubmed/24393668?dopt=Abstract</style></custom1></record></records></xml>