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 permeation 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 as by glycoproteins having an N-linked complex carbohydrate type. The apparent M(r) of intact CCA was determined to be ca 257,000 by gel filtration 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. Therefore, the intact CCA may be composed of six or eight identical subunits without disulfide bonds. In addition, CCA showed strong mitogenic activity similar to other lectins. The N-terminal amino acid of CCA may be blocked since no amino acid was detected by direct sequence analysis. Amino acid analysis showed that CCA was rich in glycine, but did not contain cysteine residues. Some properties of CCA were similar to mannose/glucose-specific legume lectins, but our data suggest that the molecular structure of CCA is different.
Nomura
K
K
Department of Plant Resource Science, Faculty of Agriculture, Kobe University, Japan. knomurak@kobe-u.ac.jp
Ashida
H
H
Uemura
N
N
Kushibe
S
S
Ozaki
T
T
Yoshida
M
M
D000372:Agglutination Tests; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008358:Mannose / Q000378:metabolism*; D051379:Mice; D008809:Mice, Inbred C3H; D008934:Mitogens / Q000494:pharmacology; D037121:Plant Lectins; D010946:Plants, Medicinal / Q000737:chemistry; D019685:Rosales / Q000737:chemistry*; D013154:Spleen / Q000166:cytology
D000596:Amino Acids; D037102:Lectins; D008934:Mitogens; D037121:Plant Lectins; D005947:Glucose; D008358:Mannose