dc.contributor.author
Gorenflos López, Jacob L.
dc.contributor.author
Dornan, Gillian L.
dc.contributor.author
Boback, Nico
dc.contributor.author
Neuenschwander, Martin
dc.contributor.author
Oder, Andreas
dc.contributor.author
Kemnitz-Hassanin, Kristin
dc.contributor.author
Schmieder, Peter
dc.contributor.author
Specker, Edgar
dc.contributor.author
Asikoglu, Hatice Ceyda
dc.contributor.author
Lauster, Daniel
dc.date.accessioned
2023-12-19T09:28:52Z
dc.date.available
2023-12-19T09:28:52Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/41790
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-41510
dc.description.abstract
Uridine diphosphate N-acetylglucosamine 2-epimerase (GNE) is a key enzyme in the sialic acid biosynthesis pathway. Sialic acids are primarily terminal carbohydrates on glycans and play fundamental roles in health and disease. In search of effective GNE inhibitors not based on a carbohydrate scaffold, we performed a high-throughput screening campaign of 68,640 drug-like small molecules against recombinant GNE using a UDP detection assay. We validated nine of the primary actives with an orthogonal real-time NMR assay and verified their IC50 values in the low micromolar to nanomolar range manually. Stability and solubility studies revealed three compounds for further evaluation. Thermal shift assays, analytical size exclusion, and interferometric scattering microscopy demonstrated that the GNE inhibitors acted on the oligomeric state of the protein. Finally, hydrogen-deuterium exchange mass spectrometry (HDX-MS) revealed which sections of GNE were shifted upon the addition of the inhibitors. In summary, we have identified three small molecules as GNE inhibitors with high potency in vitro, which serve as promising candidates to modulate sialic acid biosynthesis in more complex systems.
en
dc.format.extent
11 Seiten
dc.rights.uri
https://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject
carbohydrate biosynthesis
en
dc.subject
high-throughput screening campaign
en
dc.subject
oligomerisation inhibitors
en
dc.subject
real-time NMR
en
dc.subject
uridine diphosphate N-acetylglucosamine 2-epimerase
en
dc.subject.ddc
500 Naturwissenschaften und Mathematik::540 Chemie::540 Chemie und zugeordnete Wissenschaften
dc.title
Small Molecules Targeting Human UDP-GlcNAc 2-Epimerase
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.articlenumber
e202300555
dcterms.bibliographicCitation.doi
10.1002/cbic.202300555
dcterms.bibliographicCitation.journaltitle
ChemBioChem
dcterms.bibliographicCitation.number
24
dcterms.bibliographicCitation.volume
24
dcterms.bibliographicCitation.url
https://doi.org/10.1002/cbic.202300555
refubium.affiliation
Biologie, Chemie, Pharmazie
refubium.affiliation.other
Institut für Pharmazie
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.isPartOf.eissn
1439-7633
refubium.resourceType.provider
WoS-Alert