A normal mucin-binding lectin from the sponge Craniella australiensis

• Published in: Comparative biochemistry and physiology. Toxicology & pharmacology Vol. 143; no. 1; pp. 9 - 16
• Main Authors: Xiong, Chuannan, Li, Wei, Liu, Han, Zhang, Wei, Dou, Jiangli, Bai, Xuefang, Du, Yuguang, Ma, Xiaojun
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2006
• Subjects: Amino Acid Sequence; Animals; Cell Survival - drug effects; Cells, Cultured; Craniella australiensis; Female; Glycoproteins; Glycoproteins - pharmacology; Hemagglutination Tests; Hydrogen-Ion Concentration; Lectins - analysis; Lectins - chemistry; Lectins - isolation & purification; Lectins - metabolism; Marine; Mice; Mice, Inbred BALB C; Mitogenic; Molecular Sequence Data; Molecular Weight; Mucin-binding lectin; Mucins - metabolism; Porifera - chemistry; Spleen - cytology; Spleen - drug effects; Sponge; Temperature; Craniella australiensis; Sponge; Glycoproteins; Mitogenic; Mucin-binding lectin
• ISSN: 1532-0456; 1878-1659
• DOI: 10.1016/j.cbpc.2005.11.008

A lectin, Craniella australiensis (CAL), was isolated from sponge C. australiensis by ion-exchange on DEAE-Sephacel and purified by gel filtration on Sephadex G-150 and HPLC on DEAE-5PW. The purified lectin was a trimeric protein as revealed by SDS-PAGE and MALDI-TOF analysis. SDS-PAGE showed that the CAL protein had a molecular mass of 54 kDa, and consisted of three 18 kDa subunits. Gel filtration of purified lectin on Sephadex G-200 indicates that it exists as a 54 kDa protein in its native state. The amino acid composition was rich in Thr and Glx. CAL was found to agglutinate native and trypsinized human A, B erythrocytes, and agglutinate native erythrocytes of mouse, sheep, rabbit and chicken, and trypsinized erythrocytes of sheep and rabbit. The hemagglutination activity was inhibited by glycoproteins such as PSM and asialo-PSM, but not by any of the monosaccharides tested. The activity was stable between 20 and 70 °C. Significant CAL activity was observed between pH 5 and 8. The lectin reaction is independent of the presence of divalent cations Ca 2+ and Mg 2+. The sequence of N-terminal residues of CAL was determined as TSSCQSIVVE. The lectin showed a potent mitogenic response towards BALB/c splenocytes.