Literature DB >> 23821272

Fluorimetric and HPLC-based dengue virus protease assays using a FRET substrate.

Christoph Nitsche1, Christian D Klein.   

Abstract

The number of dengue virus infections is increasing and the dengue NS2B-NS3 protease is considered a promising target for the development of antiviral therapies. Therefore, reliable and fast screening systems are needed for the discovery of new lead structures. In this chapter, we describe two dengue virus protease assays based on an internally quenched, high-affinity Förster resonance energy transfer (FRET) substrate (Km = 105 μM). A fluorimetric assay using a microtiter fluorescence plate reader can be used for high-throughput screening of a large number of compounds. Alternatively, an HPLC-based assay with fluorescence detection can be applied to confirm the compound hits and to avoid false-positive results that may arise due to the inner filter effect of some compounds.

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Year:  2013        PMID: 23821272     DOI: 10.1007/978-1-62703-484-5_18

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  6 in total

1.  C-terminal residue optimization and fragment merging: discovery of a potent Peptide-hybrid inhibitor of dengue protease.

Authors:  Mira A M Behnam; Christoph Nitsche; Sérgio M Vechi; Christian D Klein
Journal:  ACS Med Chem Lett       Date:  2014-07-18       Impact factor: 4.345

Review 2.  Broad-spectrum agents for flaviviral infections: dengue, Zika and beyond.

Authors:  Veaceslav Boldescu; Mira A M Behnam; Nikos Vasilakis; Christian D Klein
Journal:  Nat Rev Drug Discov       Date:  2017-05-05       Impact factor: 84.694

3.  Prodrug Activation by a Viral Protease: Evaluating Combretastatin Peptide Hybrids To Selectively Target Infected Cells.

Authors:  Michael Richter; Mila M Leuthold; Dominik Graf; Ralf Bartenschlager; Christian D Klein
Journal:  ACS Med Chem Lett       Date:  2019-07-18       Impact factor: 4.345

4.  Cationic Geminoid Peptide Amphiphiles Inhibit DENV2 Protease, Furin, and Viral Replication.

Authors:  Mark Damen; Mario A Izidoro; Debora N Okamoto; Lilian C G Oliveira; Helene I V Amatdjais-Groenen; Stijn F M van Dongen; Koen W R van Cleef; Ronald P van Rij; Cindy E J Dieteren; Daniel Gironés; Bernd N M van Buuren; Byron E E Martina; Albert D M E Osterhaus; Luiz Juliano; Bob J Scholte; Martin C Feiters
Journal:  Molecules       Date:  2022-05-17       Impact factor: 4.927

5.  Kinetic characterization of trans-proteolytic activity of Chikungunya virus capsid protease and development of a FRET-based HTS assay.

Authors:  Megha Aggarwal; Rajesh Sharma; Pravindra Kumar; Manmohan Parida; Shailly Tomar
Journal:  Sci Rep       Date:  2015-10-06       Impact factor: 4.379

6.  Efficiency Improvements and Discovery of New Substrates for a SARS-CoV-2 Main Protease FRET Assay.

Authors:  Tonko Dražić; Nikos Kühl; Mila M Leuthold; Mira A M Behnam; Christian D Klein
Journal:  SLAS Discov       Date:  2021-06-19       Impact factor: 3.341

  6 in total

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