title: Purification and molecular characterization of a novel 16-kDa galectin from the nematode Caenorhabditis elegans.
abstract: In our previous study (Hirabayashi, J., Satoh, M., Ohyama, Y., and Kasai, K. (1992) J. Biochem. (Tokyo) 111, 553-555), two beta-galactoside-binding lectins (apparent subunit molecular masses, 16 and 32 kDa, respectively) were identified in the nematode Caenorhabditis elegans. The subsequent study revealed that the 32-kDa lectin is a member of the galectin family. Since the 32-kDa galectin was found to consist of two homologous domains (approximately 16 kDa), 16-kDa lectin was thought to be a degradation product of the 32-kDa galectin. To clarify this, the 16-kDa lectin was purified by an improved procedure employing extraction with a calcium-supplemented buffer. The purified 16-kDa lectin was found to exist as a dimer (approximately 30 kDa) and showed hemagglutinating activity toward trypsinized rabbit erythrocytes, which was inhibited by lactose. Almost the whole sequence of the 16-kDa polypeptide (approximately 95%, 135 amino acids) was determined after digestion with various proteases. Based on the obtained information, a full-length cDNA was cloned with the aid of RNA-polymerase chain reaction. The clone encoded 146 amino acids including initiator methionine (calculated molecular mass, 15,928 Da). Based on these results, it was concluded that the 16-kDa lectin is a novel member of the galectin family, but not a degradation product of the 32-kDa galectin as had previously thought. However, the 16-kDa galectin showed relatively low sequence similarities to both the N-terminal and the C-terminal domains of the 32-kDa galectin (28% and 27% identities, respectively) and to various vertebrate galectins (14-27%). Nonetheless, all of the critical amino acids involved in carbohydrate binding were conserved. These observations suggest that, in spite of phylogenic distance between nematodes and vertebrates, both the 16-kDa and 32-kDa nematode isolectins have conserved essentially the same function(s) as those of vertebrate galectins, probably through recognition of a key disaccharide moiety, "N-acetyllactosamine."
authors.affiliation: Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Kanagawa, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Hirabayashi
authors.forename: T
authors.initials: T
authors.lastname: Ubukata
authors.forename: K
authors.initials: K
authors.lastname: Kasai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017173:Caenorhabditis elegans / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D037161:Galectins; D006388:Hemagglutinins / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011817:Rabbits; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D037161:Galectins; D006388:Hemagglutinins