ISOLATION AND CHARACTERIZATION OF A LECTIN FROM GRIFOLA-FRONDOSA FRUITING BODIES

被引:115
作者
KAWAGISHI, H [1 ]
NOMURA, A [1 ]
MIZUNO, T [1 ]
KIMURA, A [1 ]
CHIBA, S [1 ]
机构
[1] HOKKAIDO UNIV,FAC AGR,DEPT AGR CHEM,SAPPORO,HOKKAIDO 060,JAPAN
关键词
(Grifola frondosa); Cytotoxicity; HeLa cell; Lectin; Mushroom; N-Acetyl-d-galactosamine-specific;
D O I
10.1016/0304-4165(90)90045-X
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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. © 1990.
引用
收藏
页码:247 / 252
页数:6
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