Molecular Diversity

, Volume 10, Issue 4, pp 607–618

Isolation, characterization and molecular evolution of a novel pearl shell lectin from a marine bivalve, Pteria penguin

  • Takako Naganuma
  • Tomohisa Ogawa
  • Jun Hirabayashi
  • Kenichi Kasai
  • Hisao Kamiya
  • Koji Muramoto
Full-lenth paper

DOI: 10.1007/s11030-006-9051-3

Cite this article as:
Naganuma, T., Ogawa, T., Hirabayashi, J. et al. Mol Divers (2006) 10: 607. doi:10.1007/s11030-006-9051-3


A novel lectin, PPL, was isolated from the mantle of penguin wing oyster (Pteria penguin) by affinity chromatography on mucin-Sepharose 4B and cation exchange chromatography on HiTrap SP. This lectin was estimated to be a 21-kDa monomer by gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted time of flight (MALDI-TOF) mass spectrometry. However, dynamic light scattering experiments revealed that a non-covalently linked dimer formed under high salt conditions (500 mM NaCl). Interestingly, PPL showed an increasing hemagglutinating activity with increasing salt concentration. The amino acid sequence of PPL was determined by direct protein sequence analysis and cDNA cloning. The 167-amino acid sequence included 24 lysine residues and had two tandemly repeated homologous domains (residues 20–78 and 107–165) with 44% internal homology. PPL showed sequence homology to L-rhamnose-binding lectins from fish eggs and a D-galactose-binding lectin from sea urchin eggs, with sequence identities in the range 37–48%. PPL agglutinated various animal erythrocytes independently of calcium ions. The minimum concentration of PPL needed to agglutinate rabbit erythrocytes was 0.5 µg/ml, and the most effective saccharides to inhibit the hemagglutination were D-galactose, methyl-D-galactopyranoside and N-acetyl-D-lactosamine. Lactose also inhibited hemagglutination, but L-rhamnose did so only weakly despite the sequence homology with trout egg L-rhamnose-binding lectins. The carbohydrate-binding specificity of PPL was further examined by frontal affinity chromatography using 37 different pyridylaminated oligosaccharides. PPL was found to have strong binding affinity for various oligosaccharides that have Galβ1-4Glu/GlcNAc, Galβ1-3GalNAc/GlcNAc and Galα 1-4Gal moieties in their structure. PPL had a high thermal stability and retained 50% of its hemagglutinating activity after incubation at 70°C for 100 min. It agglutinated some Gram-negative bacteria by recognizing lipopolysaccharides. Together, these results suggest that PPL is a new member of the trout egg lectin family which participates in the self-defense mechanism against bacteria and pathogens with a distinct carbohydrate-binding specificity. We conclude that the trout egg lectin family proteins, in particular their carbohydrate recognition domains, have acquired diverse carbohydrate-binding specificities during molecular evolution.


bivalveslectinPteria penguinself-defence systemthermal stability



bovine submaxillary gland mucin


cyanogen bromide,


carbohydrate recognition domain


chum salmon egg lectin


dynamic light scattering






2-(N-morpholino) ethanesulfonic acid




Pteria penguin lectin


rapid amplification of cDNA ends


l-Rhamnose-binding lectins


sea urchin egg lectin


trifluoroacetic acid


white-spotted charr egg lectin

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Takako Naganuma
    • 1
    • 2
  • Tomohisa Ogawa
    • 1
    • 2
  • Jun Hirabayashi
    • 3
  • Kenichi Kasai
    • 4
  • Hisao Kamiya
    • 5
  • Koji Muramoto
    • 1
  1. 1.Department of Biomolecular Sciences, Graduate School of Life SciencesTohoku UniversitySendaiJapan
  2. 2.Center for Interdisciplinary ResearchTohoku UniversitySendaiJapan
  3. 3.Research Center for GlycoscienceNational Institute of Advanced Industrial Science and TechnologyTsukubaJapan
  4. 4.Faculty of Pharmaceutical SciencesTeikyo UniversitySagamikoJapan
  5. 5.School of Fisheries SciencesKitasato UniversityIwateJapan