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  Protein-observed 19F NMR of LecA from Pseudomonas aeruginosa

Shanina, E., Siebs, E., Zhang, H.-X., Varón Silva, D., Joachim, I., Titz, A., et al. (2020). Protein-observed 19F NMR of LecA from Pseudomonas aeruginosa. Glycobiology, cwaa057. doi:10.1093/glycob/cwaa057.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0006-9EDF-E Version Permalink: http://hdl.handle.net/21.11116/0000-0006-C66D-1
Genre: Journal Article

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 Creators:
Shanina, Elena1, Author              
Siebs, Eike, Author
Zhang, Heng-Xi1, Author              
Varón Silva, Daniel2, Author              
Joachim, Ines, Author
Titz, Alexander, Author
Rademacher, Christoph1, Author              
Affiliations:
1Christoph Rademacher, Biomolekulare Systeme, Max Planck Institute of Colloids and Interfaces, Max Planck Society, ou_1863300              
2Daniel Varón Silva, Biomolekulare Systeme, Max Planck Institute of Colloids and Interfaces, Max Planck Society, ou_1863302              

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Free keywords: LecA, NMR, Drug discovery
 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.

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Language(s): eng - English
 Dates: 2020-06-23
 Publication Status: Published online
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: DOI: 10.1093/glycob/cwaa057
Other: oa angemahnt SN 29-06-2020
 Degree: -

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Title: Glycobiology
  Other : Glycobiology
Source Genre: Journal
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Publ. Info: Oxford : Oxford University Press
Pages: - Volume / Issue: - Sequence Number: cwaa057 Start / End Page: - Identifier: ISSN: 0959-6658