Structural Basis of the Preferential Binding for Globo-Series Glycosphingolipids Displayed by Pseudomonas aeruginosa Lectin I

Blanchard, Bertrand; Nurisso, Alessandra; Hollville, Émilie; Tétaud, Cécile; Wiels, Joëlle; Pokorná, Martina; Wimmerová, Michaela; Varrot, A.; Imberty, Anne

doi:10.1016/j.jmb.2008.08.028

Cited by 125 publications

(142 citation statements)

References 61 publications

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“…H1299 and MDCKII cells stably expressing marker proteins or Gb3 synthase were cultured in RPMI or DMEM supplemented with Geneticin and 10% (vol/vol) (5% vol/vol) FBS. Recombinant LecA-, lecA-, and non-lecA-expressing E. coli BL21 (DE3) were produced as reported (25). GFP tags and LecA mutants of P. aeruginosa PAO1 were produced as described in SI Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

“…To test the capacity of LecA as an invasion factor, we explored the invasiveness of a noninvasive E. coli strain upon induced expression of lecA by isopropyl β-D-1-thiogalactopyranoside (IPTG) (25). Remarkably, ectopic expression of lecA in E. coli BL21 (DE3) transduced this strain into an invasive one, evidenced by a significant increase of invasion into H1299 cells of about 340% compared with non-lecA-expressing E. coli strains (Fig.…”

Section: Leca Promotes a Lipid Zipper On Epithelial Cells Independentmentioning

confidence: 99%

“…The homotetrameric, galactophilic lectin LecA, which is localized to the outer bacterial membrane (21), belongs to the carbohydrate binding proteins expressed by P. aeruginosa that recognize GSLs (22,23) and represents one of the virulence factors (24). The preferential binding of LecA to the GSL globotriaosylceramide (also known as Gb3/CD77 or Pk histo-blood group antigen) (22,25) prompted us to investigate the role of LecA-Gb3 interaction in the cellular uptake of P. aeruginosa.…”

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confidence: 99%

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A lipid zipper triggers bacterial invasion

Eierhoff

Bastian

Thuenauer

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Significance Entry of bacteria into host cells critically depends on their proper engulfment by the plasma membrane. So far, actin polymerization has been described as a major driving force in this process. However, our study reveals that the interaction of the bacterial surface lectin LecA with the host cell glycosphingolipid Gb3 is fully sufficient to promote engulfment of Pseudomonas aeruginosa , whereas actin polymerization is dispensable. Hence, the formation of a “lipid zipper” represents a previously unidentified mechanism of bacterial uptake and demonstrates that bacterial pathogens have evolved lipid-based invasion strategies that may function in addition to protein receptor-based ones. Furthermore, by identifying the LecA/Gb3 interaction as the major invasion-promoting factor, our study provides new targets for drugs that may prevent bacterial invasion and dissemination.

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Section: Methodsmentioning

confidence: 99%

Section: Leca Promotes a Lipid Zipper On Epithelial Cells Independentmentioning

confidence: 99%

mentioning

confidence: 99%

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A lipid zipper triggers bacterial invasion

Eierhoff

Bastian

Thuenauer

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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123

162

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“…Materials-The recombinant protein PA-IL cloned in plasmid pET25pa1l was expressed and purified as described previously (11). The purified protein was lyophilized and stored at Ϫ20°C.…”

Section: Methodsmentioning

confidence: 99%

“…Solved crystal structures, in apo form and in complex with sugars, demonstrated that each monomer adopts a small jelly roll type ␤-sandwich fold, consisting of two curved sheets, each one formed by four antiparallel ␤-strands with a calcium ion in the binding site (10). We previously described the structural basis and thermodynamic properties of this lectin in complex with oligosaccharide moieties of glycosphingolipids combining several approaches such as cell surface labeling, glycan array analysis, titration microcalorimetry, crystallography, and molecular modeling (11). Together with previous binding studies (12,13), these data established that the globotriaosylceramide antigen Gb3, the carbohydrate moiety of which is ␣Gal1-3␤Gal1-4␤Glc, is a likely natural ligand of the lectin on human epithelia.…”

mentioning

confidence: 99%

Role of Water Molecules in Structure and Energetics of Pseudomonas aeruginosa Lectin I Interacting with Disaccharides

Nurisso

Blanchard

Audfray

et al. 2010

Journal of Biological Chemistry

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Calcium-dependent lectin I from Pseudomonas aeruginosa (PA-IL) binds specifically to oligosaccharides presenting an ␣-galactose residue at their nonreducing end, such as the disaccharides ␣Gal1-2␤GalOMe, ␣Gal1-3␤GalOMe, and ␣Gal1-4␤GalOMe. This provides a unique model for studying the effect of the glycosidic linkage of the ligands on structure and thermodynamics of the complexes by means of experimental and theoretical tools. The structural features of PA-IL in complex with the three disaccharides were established by docking and molecular dynamics simulations and compared with those observed in available crystal structures, including PA-IL⅐␣Gal1-2␤GalOMe complex, which was solved at 2.4 Å resolution and reported herein. The role of a structural bridge water molecule in the binding site of PA-IL was also elucidated through molecular dynamics simulations and free energy calculations. This water molecule establishes three very stable hydrogen bonds with O6 of nonreducing galactose, oxygen from Pro-51 main chain, and nitrogen from Gln-53 main chain of the lectin binding site. Binding free energies for PA-IL in complex with the three disaccharides were investigated, and the results were compared with the experimental data determined by titration microcalorimetry. When the bridge water molecule was included in the free energy calculations, the simulations predicted the correct binding affinity trends with the 1-2-linked disaccharide presenting three times stronger affinity ligand than the other two. These results highlight the role of the water molecule in the binding site of PA-IL and indicate that it should be taken into account when designing glycoderivatives active against P. aeruginosa adhesion.

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Structural basis of the interaction between the meningitis pathogen Streptococcus suis adhesin Fhb and its human receptor

Zhang

Hao

et al. 2016

FEBS Letters

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Edited by Peter BrzezinskiThe recently identified Streptococcus suis adhesin factor H-binding protein (Fhb) targets the host cellular receptor glycolipid GbO3 through its N terminus. However, it is unclear how Fhb interacts with its receptor. Here, we determined the complex structure of factor H-binding protein receptor-binding domain (Fhb RBD) with Gb2, an analog of its receptor, revealing that Gb2 binds in a pocket of the b sandwich core domain. We identified the key residues for Fhb RBD receptor binding using mutagenesis and isothermal titration calorimetry. Mutagenesis analyses indicated that Fhb binds to Gb2 mainly through hydrogen and hydrophobic interactions. Our findings provided structural insights into the Fhb-mediated host-pathogen interactions of S. suis.

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Structural Basis of the Preferential Binding for Globo-Series Glycosphingolipids Displayed by Pseudomonas aeruginosa Lectin I

Cited by 125 publications

References 61 publications

A lipid zipper triggers bacterial invasion

A lipid zipper triggers bacterial invasion

Role of Water Molecules in Structure and Energetics of Pseudomonas aeruginosa Lectin I Interacting with Disaccharides

Structural basis of the interaction between the meningitis pathogen Streptococcus suis adhesin Fhb and its human receptor

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