PURIFICATION AND CHARACTERIZATION OF A MANNOSE GLUCOSE-SPECIFIC LECTIN FROM CASTANEA-CRENATA/

A hemagglutinin was purified from the cotyledons of Japanese chestnut (Castanea crenata Sieb. et Zucc.) by affinity chromatography on asialo -fetuin Sepharose 4B column followed by anion-exchange and gel permeat ion chromatography. The hemagglutinating activity of Castenea crenate agglutinin (CCA) was strong for sialidase-treated human erythrocytes, but was inhibited by mannose, glucose, and their derivatives as well a s by glycoproteins having an N-linked complex carbohydrate type. The a pparent M-r of intact CCA was determined to be ca 257,000 by gel filtr ation using a Superose 12 column. In SDS-PAGE, under reducing and non- reducing conditions, CCA migrated as a single band of M-r 37,000. Ther efore, the intact CCA may be composed of six or eight identical subuni ts without disulfide bonds. In addition, CCA showed strong mitogenic a ctivity similar to other lectins. The N-terminal amino acid of CCA may be blocked since no amino acid was detected by direct sequence analys is. Amino acid analysis showed that CCA was rich in glycine, but did n ot contain cysteine residues. Some properties of CCA were similar to m annose/glucose-specific legume lectins, but our data suggest that the molecular structure of CCA is different. (C) 1998 Elsevier Science Ltd . All rights reserved.