菊芋凝集素的分离鉴定及菊芋蛋白促进家兔离体回肠蠕动的研究  

Characterization of Lectin from Helianthus tuberosus L.and Effects of Crude Extract Proteins on Rabbit-isolated Ileum

作　　者：熊川男[1] 赵小明[1] 尹恒[1] 王焕弟[1] 杜昱光[1] 

机构地区：[1]中国科学院大连化学物理研究所

出　　处：《西北农业学报》2008年第4期137-142,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：中科院知识创新重要方向项目(Kscx2-Yw-G-012)

摘　　要：经过研磨、抽提、(NH4)2SO4沉淀后得到蛋白粗提物,通过离子交换和凝胶过滤色谱对该凝集素进行了提纯。在加入和不加入2-巯基乙醇条件下,菊芋凝集素相对分子量均约为17.6 kDa;基质辅助激光解吸附电离飞行时间质谱(MALDI-TOF)检测分子量为15.379 kDa。该凝集素可以凝集兔红血球,同时也对胰蛋白酶消化过的兔红血球有凝集作用;该凝集素为Ca2+依赖型,50℃以上其凝血活力显著下降;肝素、粘蛋白和清蛋白可以抑制该凝集素的凝血活性。在家兔离体回肠试验中,胰蛋白酶裂解粗蛋白产物使平均振幅增加25%左右,胃蛋白酶裂解粗蛋白产物使平均振幅增加20%左右。试验得到的菊芋凝集素理化性质与已知的菊芋凝集素差别很大;菊芋蛋白能够促进家兔离体回肠的蠕动,酶解后的蛋白作用更强烈。Lectin from Helianthus tuberosus L. was isolated and effects of crude extract proteins on rabbit-isolated ileum were investigated. DEAD-Sephacel and Sephadex G-200 were used to purified the lectin; SDS-PAGE showed that the protein is about 17. 6kDa, MALDI TOF mass spectrometry showed the exact molecular mass was 15. 379 kDa. The lectin was found to agglutinate native and trypsinized rabbit erythrocytes. The hemagglutination activity was inhibited by heparin, PSM and albumin. The hemagglutination activity was stable below 50℃ and was depend on the presence of divalent cations Ca^2＋. The crude extract protein could act stimulation effect on rabbit-isolated ileum, and was dose-dependent. When the protein was treated with trypsin or pepsin, respectively, the products could also stimulate ileum. There are huge differences between this purified lectin and the known Heltuba from Helianthus tuberosus L. ; the crude extract protein could act stimulation effect on rabbit-isolated ileum no matter treated with or without trypsin or pepsin.

关 键 词：菊芋 凝集素 离体回肠 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学]