Purification and characterization of a T-antigen specific lectin from the coelomic fluid of a marine invertebrate, sea cucumber (Holothuria scabra)

Fish Shellfish Immunol. 2008 Apr;24(4):450-8. doi: 10.1016/j.fsi.2008.01.002. Epub 2008 Jan 11.

Abstract

A novel lectin was purified from the coelomic fluid of the sea cucumber Holothuria scabra (HSL), subjected to bacterial challenge. HSL is a monomeric glycoprotein of molecular mass 182 kDa. The lectin is highly thermostable as it retains full activity for 1 h at 80 degrees C. Further, the hemagglutination activity of HSL is unaffected by pH in the range 2-11. Unlike other lectins purified from marine invertebrates, the hemagglutination activity of HSL does not require any divalent metal ions. The affinity profile of HSL was studied by a combination of hemagglutination inhibition and fluorescence spectroscopy. HSL binds to desialylated glycoproteins, MealphaGal, T-antigen and T (alpha-ser)-antigen with a distinction between beta1-4 and beta1-3 linkages. Mealpha-T-antigen was a potent ligand having highest affinity (Ka 8.32 x 10(7)M(-1)). Monosaccharide binding is enthalphically driven while disaccharide binding involves both entropic and enthalpic contributions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / analysis
  • Animals
  • Antigens / immunology
  • Antigens / metabolism*
  • Body Fluids / chemistry*
  • Carbohydrates / analysis
  • Cations, Divalent / pharmacology
  • Circular Dichroism
  • Fluorescence
  • Hemagglutination Inhibition Tests
  • Holothuria / chemistry*
  • Holothuria / immunology
  • Hydrogen-Ion Concentration
  • Lectins / chemistry*
  • Lectins / immunology
  • Lectins / isolation & purification*
  • Lectins / metabolism
  • Mass Spectrometry
  • Molecular Weight
  • Sensitivity and Specificity
  • Temperature

Substances

  • Amino Acids
  • Antigens
  • Carbohydrates
  • Cations, Divalent
  • Lectins