Isolation and characterization of a novel fucose-binding lectin from the gill of bighead carp (Aristichthys nobilis).
Lectins play an important role in many biological systems and are used increasingly in human disease therapy. To improve our understanding of fish lectins we purified and characterized a fucose-binding lectin from the gill of bighead carp (Aristichthys nobilis). The purification procedure consisted of extracting soluble proteins in 25mM Tris-HCl buffer (pH 8.5), separation on a DEAE-Sepharose FF ion exchange column, followed by gel filtration chromatography on Sephacryl S-200 HR and Superdex 200 10/300 GL columns. The purified lectin, designated GANL, had a single protein band with an apparent molecular mass of 37kDa when subject to SDS-PAGE under reducing conditions. GANL is a homomultimeric glycoprotein with a native molecular mass of 220kDa and a carbohydrate content of approximately 13.4%. The purified lectin only agglutinated rabbit native erythrocytes, and did not require Ca(2+). Its activity was not inhibited by any of the mono- or disaccharides or glycoproteins tested, except for fucose. GANL contains a high proportion of Asp, Glu, Leu, Val, and Lys. The first 10 residues of the N-terminal region were determined as AGEQGGQCSA. The anti-microbial activity was assessed by measuring agglutination and inhibition of pathogen growth. Our results suggest that GANL agglutinates and inhibits the growth of Vibrio harveyi but has no antifungal activity.
Pan
Saikun
S
School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
Tang
Jian
J
Gu
Xiaohong
X
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000302:isolation & purification; D002347:Carps / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D029941:Fish Proteins / Q000235:genetics / Q000302:isolation & purification*; D005880:Gills / Q000276:immunology / Q000378:metabolism; D006386:Hemagglutination Tests; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D037102:Lectins / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D055550:Protein Stability; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014733:Vibrio / Q000276:immunology
D023181:Antimicrobial Cationic Peptides; D029941:Fish Proteins; D037102:Lectins; C027112:fucose-binding lectin