Chronic colonization of the lungs by opportunist bacteria such as Pseudomonas aeruginosa and members of the Bcc (Burkholderia cepacia complex) is the major cause of morbidity and mortality among CF (cystic fibrosis) patients. PA-IIL (lecB gene), a soluble lectin from Ps. aeruginosa, has been the subject of much interest because of its very strong affinity for fucose. Orthologues have been identified in the opportunist bacteria Ralstonia solanacearum, Chromobacterium violaceum and Burkholderia of Bcc. The genome of the J2315 strain of B. cenocepacia, responsible for epidemia in CF centres, contains three genes that code for proteins with PA-IIL domains. The shortest gene was cloned in Escherichia coli and pure recombinant protein, BclA (B. cenocepacia lectin A), was obtained. The presence of native BclA in B. cenocepacia extracts was checked using a proteomic approach. The specificity of recombinant BclA was characterized using surface plasmon resonance showing a preference for mannosides and supported with glycan array experiments demonstrating a strict specificity for oligomannose-type N-glycan structures. The interaction thermodynamics of BclA with methyl α-D-mannoside demonstrates a dissociation constant (Kd) of 2.75×10−6 M. The X-ray crystal structure of the complex with methyl α-D-mannoside was determined at 1.7 Å (1 Å=0.1 nm) resolution. The lectin forms homodimers with one binding site per monomer, acting co-operatively with the second dimer site. Each monomer contains two Ca2+ ions and one sugar ligand. Despite strong sequence similarity, the differences between BclA and PA-IIL in their specificity, binding site and oligomerization mode indicate that the proteins should have different roles in the bacteria.
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Research Article|
March 27 2008
Structural basis for mannose recognition by a lectin from opportunistic bacteria Burkholderia cenocepacia
Emilie Lameignere;
Emilie Lameignere
1
*CERMAV-CNRS (affiliated with Université Joseph Fourier and belonging to ICMG), BP 53, F-38041, Grenoble, Cedex 09, France
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Lenka Malinovská;
Lenka Malinovská
1
†National Centre for Biomolecular Research (NCBR) and Department of Biochemistry, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic
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Margita Sláviková;
Margita Sláviková
†National Centre for Biomolecular Research (NCBR) and Department of Biochemistry, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic
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Eric Duchaud;
Eric Duchaud
‡INRA, Unité Virologie et Immunologie Moléculaires UR892, Jouy-en-Josas, France
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Edward P. Mitchell;
Edward P. Mitchell
§ESRF Experiments Division, BP 220, F-38043, Grenoble Cedex, France
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Annabelle Varrot;
Annabelle Varrot
*CERMAV-CNRS (affiliated with Université Joseph Fourier and belonging to ICMG), BP 53, F-38041, Grenoble, Cedex 09, France
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Ondrej Šedo;
Ondrej Šedo
∥Department of Functional Genomics and Proteomics, Institute of Experimental Biology, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic
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Anne Imberty;
Anne Imberty
2
*CERMAV-CNRS (affiliated with Université Joseph Fourier and belonging to ICMG), BP 53, F-38041, Grenoble, Cedex 09, France
2Correspondence may be addressed to either of these authors (email anne.imberty@cermav.cnrs.fr or michaw@chemi.muni.cz).
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Michaela Wimmerová
Michaela Wimmerová
2
†National Centre for Biomolecular Research (NCBR) and Department of Biochemistry, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic
2Correspondence may be addressed to either of these authors (email anne.imberty@cermav.cnrs.fr or michaw@chemi.muni.cz).
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Publisher: Portland Press Ltd
Received:
September 18 2007
Revision Received:
December 20 2007
Accepted:
January 16 2008
Accepted Manuscript online:
January 16 2008
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2008 Biochemical Society
2008
Biochem J (2008) 411 (2): 307–318.
Article history
Received:
September 18 2007
Revision Received:
December 20 2007
Accepted:
January 16 2008
Accepted Manuscript online:
January 16 2008
Citation
Emilie Lameignere, Lenka Malinovská, Margita Sláviková, Eric Duchaud, Edward P. Mitchell, Annabelle Varrot, Ondrej Šedo, Anne Imberty, Michaela Wimmerová; Structural basis for mannose recognition by a lectin from opportunistic bacteria Burkholderia cenocepacia. Biochem J 15 April 2008; 411 (2): 307–318. doi: https://doi.org/10.1042/BJ20071276
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