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dc.contributor.authorShanina, Elena
dc.contributor.authorSiebs, Eike
dc.contributor.authorZhang, Hengxi
dc.contributor.authorSilva, Daniel Varón
dc.contributor.authorJoachim, Ines
dc.contributor.authorTitz, Alexander
dc.contributor.authorRademacher, Christoph
dc.date.accessioned2020-07-15T14:07:04Z
dc.date.available2020-07-15T14:07:04Z
dc.date.issued2020-06-23
dc.identifier.citationGlycobiology. 2020;cwaa057. doi:10.1093/glycob/cwaa057.en_US
dc.identifier.pmid32573695
dc.identifier.doi10.1093/glycob/cwaa057
dc.identifier.urihttp://hdl.handle.net/10033/622353
dc.description.abstractThe carbohydrate-binding protein LecA (PA-IL) from Pseudomonas aeruginosa plays an important role in the formation of biofilms in chronic infections. Development of inhibitors to disrupt LecA-mediated biofilms is desired, but limited to carbohydrate-based ligands. Moreover, discovery of drug-like ligands for LecA is challenging due to its weak affinities. Therefore, we established a protein-observed 19F (PrOF) NMR to probe ligand binding to LecA. LecA was labeled with 5 - fluoroindole to incorporate 5 - fluorotryptophanes and the resonances were assigned by site-directed mutagenesis. This incorporation did not disrupt LecA preference for natural ligands, Ca2+ and d - galactose. Following NMR resonance perturbation of W42, which is located in the carbohydrate-binding region of LecA, allowed to monitor binding of low affinity ligands such as N - acetyl d - galactosamine (d - GalNAc, Kd = 780 ± 97 μM). Moreover, PrOF NMR titration with glycomimetic of LecA p-nitrophenyl β-d-galactoside (pNPGal, Kd = 54 ± 6 μM) demonstrated a six-fold improved binding of d - Gal proving this approach to be valuable for ligand design in future drug discovery campaigns that aim to generate inhibitors of LecA.en_US
dc.language.isoenen_US
dc.publisherOxford Academicen_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjectDrug Discoveryen_US
dc.subjectLecAen_US
dc.subjectNMRen_US
dc.titleProtein-observed 19F NMR of LecA from Pseudomonas aeruginosa.en_US
dc.typeArticleen_US
dc.identifier.eissn1460-2423
dc.contributor.departmentHIPS, Helmholtz-Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1 66123 Saarbrücken, Germany.en_US
dc.identifier.journalGlycobiologyen_US
dc.source.journaltitleGlycobiology
dc.source.countryEngland


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