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To whom correspondence should be addressed:Anne Imberty, CERMAV-CNRS, 601 Rue de la Chimie, BP53, 38041 Grenoble Cedex 9, France., Tel.: 33-476037636; Fax: 33-476547203; E-mail:Imberty@cermav.cnrs.fr

We acknowledge the help received from the Cell and Tissue Imaging Core Facility of the Institut Curie (PICT-IBiSA) and the staff of the Life Imaging Center (LIC) Freiburg and the use of the advanced microscopy equipment of the LIC Freiburg and the Nikon Imaging Center at the Institut Curie-CNRS.

We also thank the MicroPiCell core facility of IFR26, Nantes, for the preparation of tissue microarrays.

Access to the High Throughput Crystallization facility of the Partnership for Structural Biology (PSB) in Grenoble was supported by the European Community PCube program.

Crystal data collection was performed at the European Synchrotron Radiation Facility, and we are grateful for the assistance in using beamline BM30A.

The glycan array resources were provided by the Consortium for Functional Glycomics (GM62116)

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Research Funding:

This work was supported by CNRS-INSERM, the German Excellence Initiative of the Deutsche Forschungsgemeinschaft, and the Association Vaincre la Mucoviscidose.

This work was also supported by Agence Nationale de la Recherche Grants NeoLect 11-BSV5-002-01 (to A. I., W. R., and J. L. P.) and GlycanClust (to W. R.) and by grants from the Czech Science Foundation (GA303/09/1168) and the CEITEC-Central European Institute of Technology (CZ.1.05/1.1.00/02.0068) from the European Regional Development Fund (to M. W.)

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Biochemistry & Molecular Biology
  • CEPACIA COMPLEX
  • PSEUDOMONAS-AERUGINOSA
  • MOLECULAR-BASIS
  • SP-NOV.
  • INFECTION
  • GLYCOSYLATION
  • POLYMORPHISM
  • SPECIFICITY
  • RECOGNITION
  • DIVERSITY

Fucose-binding Lectin from Opportunistic Pathogen Burkholderia ambifaria Binds to Both Plant and Human Oligosaccharidic Epitopes

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Journal Title:

Journal of Biological Chemistry

Volume:

Volume 287, Number 6

Publisher:

, Pages 4335-4347

Type of Work:

Article | Final Publisher PDF

Abstract:

Burkholderia ambifaria is generally associated with the rhizosphere of plants where it has biocontrol effects on other microorganisms. It is also a member of the Burkholderia cepacia complex, a group of closely related bacteria that cause lung infections in immunocompromised patients as well as in patients with granulomatous disease or cystic fibrosis. Our previous work indicated that fucose on human epithelia is a frequent target for lectins and adhesins of lung pathogens (Sulák, O., Cioci, G., Lameignére, E., Balloy, V., Round, A., Gutsche, I., Malinovská, L., Chignard, M., Kosma, P., Aubert, D. F., Marolda, C. L., Valvano, M. A., Wimmerová, M., and Imberty, A. (2011) PLoS Pathog. 7, e1002238). Analysis of the B. ambifaria genome identified BambL as a putative fucose-binding lectin. The 87- amino acid protein was produced recombinantly and demonstrated to bind to fucosylated oligosaccharides with a preference for αFuc1-2Gal epitopes. Crystal structures revealed that it associates as a trimer with two fucose-binding sites per monomer. The overall fold is a six-bladed β-propeller formed by oligomerization as in the Ralstonia solanacearum lectin and not by sequential domains like the fungal fucose lectin from Aleuria aurantia. The affinity of BambL for small fucosylated glycans is very high as demonstrated by microcalorimetry (K D < 1 μM). Plant cell wall oligosaccharides and human histo-blood group oligosaccharides H-type 2 and Lewis Y are bound with equivalent efficiency. Binding to artificial glycosphingolipid-containing vesicles, human saliva, and lung tissues confirmed that BambL could recognize a wide spectrum of fucosylated epitopes, albeit with a lower affinity for biological material from nonsecretor individuals.

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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