红花石蒜球茎凝集素的纯化及部分性质  

PURIFICATION AND CHARACTERIZATION OF AGGLUTININ FROMBULBS OF LYCORIS RADIATA (AMARYLLIDACEAE)

作　　者：常丽青[1] 吴传芳[1] 吕鸿周[1] 刘超[1] 顾莹[1] 陈放[1] 吴洽庆[2] 鲍锦库[1] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]中国科学院成都生物研究所,成都610041

出　　处：《应用与环境生物学报》2005年第2期164-167,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金资助项目 (No. 30000032 )~~

摘　　要：石蒜科植物红花石蒜(Lycoris radiata)球茎经生理盐水浸取、硫酸铵分级沉淀、DEAE-Sepharose和SephacrylS-100分子筛层析得到石蒜凝集素(L.radiata agglutinin,LRA).经SDS-PAGE检测为单一蛋白带,亚基相对分子量Mr为12×103,Sephacryl S-100凝胶过滤测定Mr约为45×103,表明LRA是由4个相同亚基组成的蛋白.LRA能凝集兔红细胞和大肠杆菌,最低凝集浓度分别为0.95μg/mL和1.9μg/mL;LRA还能凝集啤酒酵母细胞.糖抑制实验显示,甘露聚糖和甲状腺球蛋白能有效抑制LRA的凝血活性.促淋巴细胞有丝分裂试验结果表明,LRA对外周血淋巴细胞具有很强的促有丝分裂能力;同时,LRA能有效地降低Ⅱ型人类单纯疱疹病毒(HSV-Ⅱ)对Vero细胞(非洲绿猴肾细胞)的感染,ρ(IC50)=5～10μg/mL之间,并且在500μg/mL时对正常Vero细胞无细胞毒性.Lycoris radiata agglutinin (LRA) was isolated from the bulbs of L. radiata by extraction, precipitation with (NH 4 ) 2 SO 4 , ion-exchange chromatography on DEAE-Sepharose and gel filtration on Sephacryl S-100. The purified lectin showed a single protein band on SDS-PAGE with the subunit molecular weight of 12×10 3 . The molecular weight of LRA was 45×10 3 determined by gel filtration on Sephacryl S-100. LRA could agglutinate rabbit erythrocyte at 0.95 μg/mL and E. coli cells at 1.9 μg/mL, and it could also agglutinate Saccharomyces cerevisiae cells. The result of carbohydrate inhibition assay showed that Mannan and Thyroglobulin could inhibit the agglutinating activity of LRA. LRA had strong mitogenetic activity toward human T-lymphocyte. LRA showed antivirus activity to Herpes simplex virus II (HSV-II), and the IC 50 was 5~10 μg/mL. It showed no cytotoxicity toward vero cells even at 500 μg/mL. Fig 1, Tab 6, Ref 15

关 键 词：石蒜凝集素(LRA) 纯化 凝集活性 促淋巴细胞有丝分裂活性 抗病毒活性 

分 类 号：Q946.1[生物学—植物学]