Structure, properties and enhanced expression of galactose-binding C-type lectins in mucous cells of gills from freshwater Japanese eels (Anguilla japonica).
Using a Japanese-eel (Anguilla japonica) gill cDNA subtraction library, two novel beta-d-galactose-binding lectins were identified that belong to group VII of the animal C-type lectin family. The eel C-type lectins, termed eCL-1 and eCL-2, are simple lectins composed of 163 amino acid residues, including a 22-residue signal peptide for secretion and a single carbohydrate-recognition domain (CRD) of approximately 130 residues typical of C-type lectins. The galactose specificity of the CRD was suggested by the presence of a QPD motif and confirmed by a competitive binding assay. Using Ruthenium Red staining, the lectins were shown to bind Ca(2+) ions. SDS/PAGE showed that native eCL-1 and eCL-2 have an SDS-resistant octameric structure (a tetramer of disulphide-linked dimers). Northern and Western blot analyses demonstrated high-level expression of eCL-1 and eCL-2 mRNAs and their protein products in gills from freshwater eels, which decreased markedly when the eels were transferred from freshwater to seawater. Immunohistochemistry showed that the eel lectins are localized in the exocrine mucous cells of the gill.
Mistry
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AC
Department of Biological Sciences, Tokyo Institute of Technology, Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Honda
S
S
Hirose
S
S
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000811:Anguilla; D000818:Animals; D001212:Asialoglycoproteins / Q000737:chemistry; D001483:Base Sequence; D001665:Binding Sites; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D060748:Fetuins; D037161:Galectins; D015723:Gene Library; D005880:Gills / Q000378:metabolism*; D006388:Hemagglutinins / Q000096:biosynthesis* / Q000737:chemistry*; D007150:Immunohistochemistry; D037102:Lectins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012430:Ruthenium Red / Q000494:pharmacology; D012685:Sepharose / Q000737:chemistry; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors; D014018:Tissue Distribution; D000509:alpha-Fetoproteins / Q000737:chemistry
D000596:Amino Acids; D001212:Asialoglycoproteins; D018076:DNA, Complementary; D060748:Fetuins; D037161:Galectins; D006388:Hemagglutinins; D037102:Lectins; D012333:RNA, Messenger; D000509:alpha-Fetoproteins; C017029:asialofetuin; D012430:Ruthenium Red; D012313:RNA; D012685:Sepharose