大杯伞凝集素的分离纯化及其性质  

ISOLATION, PURIFICATION AND CHARACTERIZATION OF LECTIN FROM CLITOCYBE MAXIMA

作　　者：林玉满[1] 苏爱华 

机构地区：[1]福建师范大学化学与材料学院,福州350007 [2]福建卫生学校,福州350001

出　　处：《应用与环境生物学报》2004年第4期497-501,共5页Chinese Journal of Applied and Environmental Biology

基　　金：福建省高校测试基金资助项目 (CJ - 0 57)~~

摘　　要：大杯伞子实体经生理盐水抽提、硫酸铵分步沉淀、DEAE -Sepharose和SephadexG - 10 0柱层析纯化得到大杯伞凝集素 (Clitocybemaximalectin ,CML) .CML经PAGE显示单一条带 ,SDS -PAGE测得其亚基相对分子质量为4 9× 10 3 ,SephadexG - 10 0凝胶过滤测得相对分子质量为 4 9× 10 3 ,CML不含糖 ,等电点为 4 .82 .该凝集素对小鼠脾脏淋巴细胞、小鼠S180 肉瘤细胞和人的A、B、O和AB型血细胞具有凝集作用 ,对兔红细胞的凝集作用可被半乳糖 (galac tose)所抑制 .氨基酸组成分析表明 ,CML含有 16种氨基酸 ,其中天冬氨酸 (asparticacid)、谷氨酸 (glutamicacid)和亮氨酸 (leucine)含量较高 .CML对热、酸碱具有一定的稳定性 ,经 6 0℃处理 10min ,仍有较高的活性 ,在pH 5～ 8范围内较稳定 ,其凝血活性受Mg2 + 、Ca2 + 、Zn2 + 和Mn2 + 二价阳离子的影响 .CML对小鼠腹腔注射的半致死量为 2 5 .70mg/kg.图 4表 7参CML from the fruiting bodies of Clitocybe maxima (Gartn et Mey.ex Fr)Quel was isolated and purified via physiological saline extraction, precipitation of 30%～80% (NH 4) 2SO 4 and DEAE-Sepharose, Sephadex G-100 chromatography. It turned out to be a single band in PAGE. SDS-PAGE showed that it contained one subunit, with relative molecular mass 49×103. The relative molecular mass of CML was 49×103 tested by gel filtration. Its pI was about 4.82 by isoelectric focusing. This lectin agglutinated rabbit erythrocytes, human erythrocytes, the peripheral lymphocytes cells and S 180 tumor cells from mouse. The hemagglutination of the lectin on rabbit erythrocytes was inhibited by galactose among the sugar tested. Amino acid analysis showed that it contained 16 kinds of amino acids, in which the contents of aspartic acid, glutamic acid and leucine were relatively high. CML has certain thermal-, acid- and alkali-stablity. It still possesseds partial activity after being heated 10 minutes under 60 ℃, and high activity within pH 5～8. Its activity was affected by demetalization and addition of divalent metals including Mg 2+ ,Ca 2+,Zn 2+ and Mn 2+. CML was toxic to mouse by celiac injection with an LD 50 of 25.70 mg/kg. Fig 4, Tab 7, Ref 14

关 键 词：凝集素 大杯伞 分离纯化 性质 

分 类 号：Q946[生物学—植物学]