Purification and characterization of four Ca(2+)-dependent lectins from the marine invertebrate, Cucumaria echinata

• Published in: Journal of biochemistry (Tokyo) Vol. 116; no. 1; pp. 209 - 214
• Main Authors: Hatakeyama, T, Kohzaki, H, Nagatomo, H, Yamasaki, N
• Format: Journal Article
• Language: English
• Published: England 01.07.1994
• Subjects: Animals; Calcium - pharmacology; Carbohydrates - pharmacology; Erythrocytes - metabolism; Hemagglutination Tests; Humans; Lectins - isolation & purification; Molecular Weight; Sea Cucumbers - chemistry
• ISSN: 0021-924X

Four Ca(2+)-dependent, N-acetylgalactosamine/galactose-specific lectins were purified from the marine invertebrate, Cucumaria echinata (Holothuroidea), by column chromatography on lactosyl-Sepharose 4B, Sephacryl S-200, and Q-Sepharose. The molecular masses of these lectins were estimated to be 27 kDa (CEL-I), 35 kDa (CEL-II), 45 kDa (CEL-III), and 68 kDa (CEL-IV) on SDS-PAGE under nonreducing conditions. Among these lectins, CEL-I and CEL-IV strongly agglutinated rabbit and human erythrocytes, and were found to recognize N-acetylgalactosamine and galactose-containing carbohydrates from the results of a hemagglutination inhibition assay. In contrast, CEL-II failed to agglutinate any erythrocytes tested, although its carbohydrate-binding ability was confirmed by a carbohydrate-binding assay involving asialofetuin-horseradish peroxidase. Interestingly, CEL-III caused hemolysis of rabbit and human erythrocytes, while it showed only hemagglutination of chicken and horse erythrocytes at relatively high concentrations. The hemolytic activity of CEL-III was also dependent on the Ca(2+)-concentration, and inhibited by N-acetylgalactosamine and galactose-containing carbohydrates, suggesting that the hemolysis was caused by Ca(2+)-dependent binding of CEL-III to specific carbohydrate chains on the erythrocyte surface and the following partial destruction of the membrane.