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2020-11-09Zeitschriftenartikel DOI: 10.1002/chem.202003283
Liposomal FRET Assay Identifies Potent Drug‐Like Inhibitors of the Ceramide Transport Protein (CERT)
dc.contributor.authorSamaha, Doaa
dc.contributor.authorHamdo, Housam H.
dc.contributor.authorCong, Xiaojing
dc.contributor.authorSchumacher, Fabian
dc.contributor.authorBanhart, Sebastian
dc.contributor.authorAglar, Öznur
dc.contributor.authorMöller, Heiko M.
dc.contributor.authorHeuer, Dagmar
dc.contributor.authorKleuser, Burkhard
dc.contributor.authorM. Saied, Essa
dc.contributor.authorArenz, Christoph
dc.date.accessioned2021-05-06T11:06:39Z
dc.date.available2021-05-06T11:06:39Z
dc.date.issued2020-11-09none
dc.date.updated2021-02-15T09:51:19Z
dc.identifier.issn0947-6539
dc.identifier.urihttp://edoc.hu-berlin.de/18452/23539
dc.description.abstractCeramide transfer protein (CERT) mediates non‐vesicular transfer of ceramide from endoplasmic reticulum to Golgi apparatus and thus catalyzes the rate‐limiting step of sphingomyelin biosynthesis. Usually, CERT ligands are evaluated in tedious binding assays or non‐homogenous transfer assays using radiolabeled ceramides. Herein, a facile and sensitive assay for CERT, based on Förster resonance energy transfer (FRET), is presented. To this end, we mixed donor and acceptor vesicles, each containing a different fluorescent ceramide species. By CERT‐mediated transfer of fluorescent ceramide, a FRET system was established, which allows readout in 96‐well plate format, despite the high hydrophobicity of the components. Screening of a 2 000 compound library resulted in two new potent CERT inhibitors. One is approved for use in humans and one is approved for use in animals. Evaluation of cellular activity by quantitative mass spectrometry and confocal microscopy showed inhibition of ceramide trafficking and sphingomyelin biosynthesis.eng
dc.description.sponsorshipDeutsche Forschungsgemeinschaft http://dx.doi.org/10.13039/501100001659
dc.language.isoengnone
dc.publisherHumboldt-Universität zu Berlin
dc.rights(CC BY 4.0) Attribution 4.0 Internationalger
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectenzyme assayseng
dc.subjectFörster resonance energy transfer (FRET)eng
dc.subjectliposomeseng
dc.subjectsphingolipidseng
dc.subjecttransport proteinseng
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftennone
dc.titleLiposomal FRET Assay Identifies Potent Drug‐Like Inhibitors of the Ceramide Transport Protein (CERT)none
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:11-110-18452/23539-7
dc.identifier.doi10.1002/chem.202003283none
dc.identifier.doihttp://dx.doi.org/10.18452/22862
dc.type.versionpublishedVersionnone
local.edoc.container-titleChemistry – a European journalnone
local.edoc.pages6none
local.edoc.type-nameZeitschriftenartikel
local.edoc.institutionMathematisch-Naturwissenschaftliche Fakultätnone
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-publisher-nameWiley-VCHnone
local.edoc.container-publisher-placeWeinheimnone
local.edoc.container-volume26none
local.edoc.container-issue70none
local.edoc.container-firstpage16616none
local.edoc.container-lastpage16621none
dc.description.versionPeer Reviewednone
dc.identifier.eissn1521-3765
local.edoc.affiliationSamaha, Doaa: Insitute for Chemistry, Humboldt Universität zu Berlin, Brook-Taylor-Strasse 2 12489 Berlin Germanynone
local.edoc.affiliationHamdo, Housam H.: Insitute for Chemistry, Humboldt Universität zu Berlin, Brook-Taylor-Strasse 2 12489 Berlin Germanynone
local.edoc.affiliationCong, Xiaojing, CNRS, Institut de Chimie de Nice, Université Côte d'Azur, 06108 Nice Francenone
local.edoc.affiliationSchumacher, Fabian: Department of Toxicology, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114–116 14558 Nuthetal Germanynone
local.edoc.affiliationBanhart, Sebastian: Unit 'Sexually Transmitted Bacterial Infections', Department of Infectious Diseases, Robert Koch Institute, 13353 Berlin Germanynone
local.edoc.affiliationAglar, Öznur: Universität Potsdam, Institut für Chemie, Karl-Liebknecht-Strasse 24–25, Haus 25 14476 Golm Germanynone
local.edoc.affiliationMöller, Heiko M.: Universität Potsdam, Institut für Chemie, Karl-Liebknecht-Strasse 24–25, Haus 25 14476 Golm Germanynone
local.edoc.affiliationHeuer, Dagmar: Unit 'Sexually Transmitted Bacterial Infections', Department of Infectious Diseases, Robert Koch Institute, 13353 Berlin Germanynone
local.edoc.affiliationKleuser, Burkhard: Department of Toxicology, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114–116 14558 Nuthetal Germanynone
local.edoc.affiliationSaied, Essa M.: Insitute for Chemistry, Humboldt Universität zu Berlin, Brook-Taylor-Strasse 2 12489 Berlin Germanynone
local.edoc.affiliationArenz, Christoph: Insitute for Chemistry, Humboldt Universität zu Berlin, Brook-Taylor-Strasse 2 12489 Berlin Germanynone

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