Glycoconjugate Journal

, Volume 11, Issue 6, pp 507–517

The monosaccharide binding site of lentil lectin: an X-ray and molecular modelling study

  • Remy Loris
  • Florence Casset
  • Julie Bouckaert
  • Jurgen Pletinckx
  • Minh-Hoa Dao-Thi
  • Freddy Poortmans
  • Anne Imberty
  • Serge Perez
  • Lode Wyns
Lectin Papers

Abstract

The X-ray crystal structure of lentil lectin in complex with α-d-glucopyranose has been determined by molecular replacement and refined to anR-value of 0.20 at 3.0 Å resolution. The glucose interacts with the protein in a manner similar to that found in the mannose complexes of concanavalin A, pea lectin and isolectin I fromLathyrus ochrus. The complex is stabilized by a network of hydrogen bonds involving the carbohydrate oxygens O6, O4, O3 and O5. In addition, the α-d-glucopyranose residue makes van der Waals contacts with the protein, involving the phenyl ring of Phe123β. The overall structure of lentil lectin, at this resolution, does not differ significantly from the highly refined structures of the uncomplexed lectin.

Molecular docking studies were performed with mannose and its 2-O and 3-O-m-nitro-benzyl derivatives to explain their high affinity binding. The interactions of the modelled mannose with lentil lectin agree well with those observed experimentally for the protein-carbohydrate complex. The highly flexible Me-2-O-(m-nitro-benzyl)-α-d-mannopyranoside and Me-3-O-(m-nitro-benzyl)-α-d-mannopyranoside become conformationally restricted upon binding to lentil lectin. For best orientations of the two substrates in the combining site, the loss of entropy is accompanied by the formation of a strong hydrogen bond between the nitro group and one amino acid, Gly97β and Asn125β, respectively, along with the establishment of van der Waals interactions between the benzyl group and the aromatic amino acids Tyr100β and Trp128β.

Keywords

lectin lentil lectin carbohydrate specificity glucose binding mannose binding 

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Copyright information

© Chapman & Hall 1994

Authors and Affiliations

  • Remy Loris
    • 1
  • Florence Casset
    • 2
  • Julie Bouckaert
    • 1
  • Jurgen Pletinckx
    • 1
  • Minh-Hoa Dao-Thi
    • 1
  • Freddy Poortmans
    • 3
  • Anne Imberty
    • 4
  • Serge Perez
    • 2
  • Lode Wyns
    • 1
  1. 1.Laboratorium voor Ultrastructuur, Instituut voor Moleculaire BiologieVrije Universiteit BrusselSint-Genesius-RodeBelgium
  2. 2.Ingenierie Moléculaire, INRANantes cedex 03France
  3. 3.Vlaamse Instelling voor Technologisch Onderzoek—VITOMolBelgium
  4. 4.LSO-CNRS, Faculté des Sciences et TechniquesNantes cedex 03France

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