Gutenberg Open Science: A competition smFRET assay to study ligand-induced conformational changes of the dengue virus protease

Please use this identifier to cite or link to this item: http://doi.org/10.25358/openscience-10096

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DC Field	Value	Language
dc.contributor.author	Maus, Hannah	-
dc.contributor.author	Hinze, Gerald	-
dc.contributor.author	Hammerschmidt, Stefan Josef	-
dc.contributor.author	Basché, Thomas	-
dc.contributor.author	Schirmeister, Tanja	-
dc.date.accessioned	2024-02-16T08:34:18Z	-
dc.date.available	2024-02-16T08:34:18Z	-
dc.date.issued	2023	-
dc.identifier.uri	https://openscience.ub.uni-mainz.de/handle/20.500.12030/10114	-
dc.description.abstract	Ligand binding to proteins often is accompanied by conformational transitions. Here, we describe a competition assay based on single molecule Förster resonance energy transfer (smFRET) to investigate the ligand-induced conformational changes of the dengue virus (DENV) NS2B-NS3 protease, which can adopt at least two different conformations. First, a competitive ligand was used to stabilize the closed conformation of the protease. Subsequent addition of the allosteric inhibitor reduced the fraction of the closed conformation and simultaneously increased the fraction of the open conformation, demonstrating that the allosteric inhibitor stabilizes the open conformation. In addition, the proportions of open and closed conformations at different concentrations of the allosteric inhibitor were used to determine its binding affinity to the protease. The KD value observed is in accordance with the IC50 determined in the fluorometric assay. Our novel approach appears to be a valuable tool to study conformational transitions of other proteases and enzymes.	en_GB
dc.language.iso	eng	de
dc.rights	CC BY-NC-ND	*
dc.rights.uri	https://creativecommons.org/licenses/by-nc-nd/4.0/	*
dc.subject.ddc	540 Chemie	de_DE
dc.subject.ddc	540 Chemistry and allied sciences	en_GB
dc.subject.ddc	610 Medizin	de_DE
dc.subject.ddc	610 Medical sciences	en_GB
dc.title	A competition smFRET assay to study ligand-induced conformational changes of the dengue virus protease	en_GB
dc.type	Zeitschriftenaufsatz	de
dc.identifier.doi	http://doi.org/10.25358/openscience-10096	-
jgu.type.contenttype	Scientific article	de
jgu.type.dinitype	article	en_GB
jgu.type.version	Published version	de
jgu.type.resource	Text	de
jgu.organisation.department	FB 09 Chemie, Pharmazie u. Geowissensch.	de
jgu.organisation.number	7950	-
jgu.organisation.name	Johannes Gutenberg-Universität Mainz	-
jgu.rights.accessrights	openAccess	-
jgu.journal.title	Protein science	de
jgu.journal.volume	32	de
jgu.journal.issue	1	de
jgu.pages.alternative	4526	de
jgu.publisher.year	2023	-
jgu.publisher.name	Wiley	de
jgu.publisher.place	Hoboken, NJ	de
jgu.publisher.issn	1469-896X	de
jgu.organisation.place	Mainz	-
jgu.subject.ddccode	540	de
jgu.subject.ddccode	610	de
jgu.publisher.doi	10.1002/pro.4526	de
jgu.organisation.ror	https://ror.org/023b0x485	-
jgu.subject.dfg	Naturwissenschaften	de
Appears in collections:	DFG-491381577-H

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