Protein-observed 19F NMR of LecA from Pseudomonas aeruginosa.
LecA
NMR
drug discovery
lectin
Journal
Glycobiology
ISSN: 1460-2423
Titre abrégé: Glycobiology
Pays: England
ID NLM: 9104124
Informations de publication
Date de publication:
09 02 2021
09 02 2021
Historique:
received:
02
04
2020
revised:
05
06
2020
accepted:
16
06
2020
pubmed:
24
6
2020
medline:
22
2
2022
entrez:
24
6
2020
Statut:
ppublish
Résumé
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 it is limited to carbohydrate-based ligands. Moreover, discovery of drug-like ligands for LecA is challenging because of its weak affinities. Therefore, we established a protein-observed 19F (PrOF) nuclear magnetic resonance (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 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 6-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.
Identifiants
pubmed: 32573695
pii: 5861409
doi: 10.1093/glycob/cwaa057
pmc: PMC7874386
doi:
Substances chimiques
Adhesins, Bacterial
0
LecA protein, bacteria
0
Recombinant Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
159-165Informations de copyright
© The Author(s) 2020. Published by Oxford University Press.
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