Isolation and characterization of a lectin from Grifola frondosa fruiting bodies.
An N-acetylgalactosamine-specific lectin (GFL) was isolated from Grifola frondosa fruiting bodies by affinity chromatographies on acid-treated Sepharose CL-4B and then GalNAc-Toyopearl. The isolated lectin agglutinated all types of erythrocytes equally. Molecular masses estimated by gel filtration under various buffers and matrices varied from 30 to 52 kDa. On the other hand, SDS-PAGE in the presence or absence of 2-mercaptoethanol showed three major bands of 33, 66 and 100 kDa and a faint band of 65 kDa. This lectin exhibited GalNAc-specificity. The protein was a glycoprotein containing 3.3% total sugar, and the amino acid analysis revealed a high content of acidic and hydroxy amino acids and a low content of methionine and histidine. GFL was cytotoxic against HeLa cells. The toxicity did not appear after preincubating the lectin with the haptenic sugar N-acetylgalactosamine.
Kawagishi
H
H
Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Japan.
Nomura
A
A
Mizuno
T
T
Kimura
A
A
Chiba
S
S
D000116:Acetylgalactosamine / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D001487:Basidiomycota / Q000032:analysis*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000494:pharmacology; D002470:Cell Survival; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D037102:Lectins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D008623:Mercaptoethanol; D008969:Molecular Sequence Data; D008970:Molecular Weight
D000596:Amino Acids; D002241:Carbohydrates; D037102:Lectins; D008623:Mercaptoethanol; D000116:Acetylgalactosamine