Protein-observed 19F NMR of LecA from Pseudomonas aeruginosa.
dc.contributor.author | Shanina, Elena | |
dc.contributor.author | Siebs, Eike | |
dc.contributor.author | Zhang, Hengxi | |
dc.contributor.author | Silva, Daniel Varón | |
dc.contributor.author | Joachim, Ines | |
dc.contributor.author | Titz, Alexander | |
dc.contributor.author | Rademacher, Christoph | |
dc.date.accessioned | 2020-07-15T14:07:04Z | |
dc.date.available | 2020-07-15T14:07:04Z | |
dc.date.issued | 2020-06-23 | |
dc.identifier.citation | Glycobiology. 2020;cwaa057. doi:10.1093/glycob/cwaa057. | en_US |
dc.identifier.pmid | 32573695 | |
dc.identifier.doi | 10.1093/glycob/cwaa057 | |
dc.identifier.uri | http://hdl.handle.net/10033/622353 | |
dc.description.abstract | The 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.iso | en | en_US |
dc.publisher | Oxford Academic | en_US |
dc.rights | Attribution-NonCommercial-ShareAlike 4.0 International | * |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | * |
dc.subject | Drug Discovery | en_US |
dc.subject | LecA | en_US |
dc.subject | NMR | en_US |
dc.title | Protein-observed 19F NMR of LecA from Pseudomonas aeruginosa. | en_US |
dc.type | Article | en_US |
dc.identifier.eissn | 1460-2423 | |
dc.contributor.department | HIPS, Helmholtz-Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1 66123 Saarbrücken, Germany. | en_US |
dc.identifier.journal | Glycobiology | en_US |
dc.source.journaltitle | Glycobiology | |
dc.source.country | England |