Isolation, purification, characterization and glycan-binding profile of a d-galactoside specific lectin from the marine sponge, Halichondria okadai

• Published in: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2008-08, Vol.150 (4), p.349-357
• Main Authors: Kawsar, Sarkar M.A., Fujii, Yuki, Matsumoto, Ryo, Ichikawa, Takayuki, Tateno, Hiroaki, Hirabayashi, Jun, Yasumitsu, Hidetaro, Dogasaki, Chikaku, Hosono, Masahiro, Nitta, Kazuo, Hamako, Jiharu, Matsui, Taei, Ozeki, Yasuhiro
• Format: Article
• Language: English
• Subjects: Acetylgalactosamine - pharmacology; alpha-Fetoproteins - pharmacology; Animals; Asialoglycoproteins - pharmacology; Carbohydrate Sequence; Cations, Divalent - pharmacology; Chromatography, Affinity; Dansyl Compounds - pharmacology; Electrophoresis, Polyacrylamide Gel; Fetuins; Frontal affinity chromatography; Galactosamine - analogs & derivatives; Galactosamine - pharmacology; Galactose - pharmacology; Galectin; Galectins - isolation & purification; Galectins - metabolism; Galectins - pharmacology; Glycosides - metabolism; Halichondria okadai; Hemagglutination - drug effects; Humans; Isoelectric Focusing; Lactose - pharmacology; Melibiose - pharmacology; Methylgalactosides - pharmacology; Molecular Sequence Data; Porifera - metabolism; Primary structure; Rabbits; Sponge lectin
• ISSN: 1096-4959; 1879-1107
• DOI: 10.1016/j.cbpb.2008.04.004

A lectin recognizing both Galβ1-3GlcNAc and Galβ1-4GlcNAc was purified from the demosponge Halichondria okadai by lactosyl-agarose affinity chromatography. The molecular mass of the lectin was determined to be 30 kDa by SDS-PAGE under reducing and non-reducing conditions and 60 kDa by gel permeation chromatography. The p I value of the lectin was 6.7. It was found to agglutinate trypsinized and glutaraldehyde-fixed rabbit and human erythrocytes in the presence and absence of divalent cations. The hemagglutinating activity by the lectin was inhibited by d-galactose, methyl- d-galactopyranoside, N-acetyl- d-galactosamine, methyl- N-acetyl- d-galactosaminide, lactose, melibiose, and asialofetuin. The K d of the lectin against p-nitrophenyl-β-lactoside was determined to be 2.76 × 10 − 5  M and its glycan-binding profile given by frontal affinity chromatography was shown to be similar to many other known galectins. Partial primary structure analysis of 7 peptides by cleavage with lysyl endopeptidase indicated that one of the peptides showed significant similarity with galectin purified from the sponge Geodia cydonium.