A fucose-binding superlectin from Enterobacter cloacae with high Lewis and ABO blood group antigen specificity

authored by: Ghamdan Beshr, Asfandyar Sikandar, Julia Gläser, Mario Fares, Roman Sommer, Stefanie Wagner, Jesko Köhnke, Alexander Titz
Abstract: Bacteria frequently employ carbohydrate-binding proteins, so-called lectins, to colonize and persist in a host. Thus, bacterial lectins are attractive targets for the development of new anti-infectives. To find new potential targets for anti-infectives against pathogenic bacteria, we searched for homologs of Pseudomonas aeruginosa lectins and identified homologs of LecA in Enterobacter species. Here, we recombinantly produced and biophysically characterized a homolog that comprises one LecA domain and one additional, novel protein domain. This protein was termed Enterobacter cloacae lectin A (EclA) and found to bind L-fucose. Glycan array analysis revealed a high specificity for the LewisA antigen and the type II H-antigen (blood group O) for EclA, while related antigens LewisX, Y, and B, as well as blood group A or B were not bound. We developed a competitive binding assay to quantify blood group antigen-binding specificity in solution. Finally, the crystal structure of EclA could be solved in complex with methyl α-L-selenofucoside. It revealed the unexpected binding of the carbohydrate ligand to the second domain, which comprises a novel fold that dimerizes via strand-swapping resulting in an intertwined beta sheet.
Organisation(s): Institute of Food Chemistry
External Organisation(s): Helmholtz Centre for Infection Research (HZI)
German Center for Infection Research (DZIF)
Saarland University
Type: Article
Journal: Journal of Biological Chemistry
Volume: 301
ISSN: 0021-9258
Publication date: 02.2025
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Biochemistry, Molecular Biology, Cell Biology
Electronic version(s): https://doi.org/10.1016/j.jbc.2024.108151 (Access: Open)

BibTeX

@article{fd5c0631878549b79bab46bf746183d6,
title = "A fucose-binding superlectin from Enterobacter cloacae with high Lewis and ABO blood group antigen specificity",
abstract = "Bacteria frequently employ carbohydrate-binding proteins, so-called lectins, to colonize and persist in a host. Thus, bacterial lectins are attractive targets for the development of new anti-infectives. To find new potential targets for anti-infectives against pathogenic bacteria, we searched for homologs of Pseudomonas aeruginosa lectins and identified homologs of LecA in Enterobacter species. Here, we recombinantly produced and biophysically characterized a homolog that comprises one LecA domain and one additional, novel protein domain. This protein was termed Enterobacter cloacae lectin A (EclA) and found to bind L-fucose. Glycan array analysis revealed a high specificity for the LewisA antigen and the type II H-antigen (blood group O) for EclA, while related antigens LewisX, Y, and B, as well as blood group A or B were not bound. We developed a competitive binding assay to quantify blood group antigen-binding specificity in solution. Finally, the crystal structure of EclA could be solved in complex with methyl α-L-selenofucoside. It revealed the unexpected binding of the carbohydrate ligand to the second domain, which comprises a novel fold that dimerizes via strand-swapping resulting in an intertwined beta sheet.",
keywords = "adhesion, blood group, carbohydrates, Enterobactercloacae, H-type antigen, LecA, lectin, LewisA",
author = "Ghamdan Beshr and Asfandyar Sikandar and Julia Gl{\"a}ser and Mario Fares and Roman Sommer and Stefanie Wagner and Jesko K{\"o}hnke and Alexander Titz",
note = "Publisher Copyright: {\textcopyright} 2024 The Authors",
year = "2025",
month = feb,
doi = "10.1016/j.jbc.2024.108151",
language = "English",
volume = "301",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "2",
}