雷丸凝集素的纯化及理化性质的研究  

STUDIES ON THE PURIFICATION AND PROPERTIES OF OMPHALIA LAPIDESCENS LECTIN

作　　者：于勇海[1] 龚隽[1] 余明琨[1] 

机构地区：[1]中国科学技术大学研究生院生物教学部,北京100039

出　　处：《菌物系统》2000年第2期278-282,共5页Mycosystema

摘　　要：雷丸（OmphalialapidescensSchroct．）经过Tris－HCl缓冲液浸提，硫酸铵分级沉淀，离子交换，卵粘蛋白－Sepharose4B亲和层析以及Sephacryl-S100分子筛等步骤，纯化得到纯度为95％以上的雷扎凝集素（简称OLL）。比活提高45.8倍，活力回收2.5％。雷丸凝集素是单一肽链的蛋白质，分子量为12kDa，等电点为7.5，可被半乳糖抑制。具有热稳定性及酸碱（pH1～12）稳定性。2mmol／L氯化锌可使其比活提高4倍。同兔血相比，与人血进行凝集反应，其比活可提高4倍。进行圆二光谱测定，α－螺旋和β-折叠含量较高。与半乳糖间的抑制反应瞬间内完成，且空间结构变得紧密。Omphalia lapideacens lectin (OLL) has been purified by extraction with Tris-HCl buffer,anunonium sulfate frachonation, ion-exchange column, and affinity chromatograPhy on an ovomucoid-Sepharose 4B column and Sephacryl-S100 column. The purified OLL shows an achvity increase of 45.8 fold, an achvity recovery of 2.5% and purity of above 95%. The hemagglutinating activity of OLL on rabbit red blood cel1s was inhibited by D-galactose. The molecular weight of OLL was 12kD OLL was heat stable and it was comparahvely stable at pH values from 1 to 12. Zinc Chioride (2mmol / L) incfeasedit,s hemapgluhnating achvity by 4 fold and the hemagglutinahng activity of OLL on human red blood cells was increased by 4 fold. Cireular dichroism spectra revealed high content of a-helix and -sheet structures in this lectin. From the spectrum observed in the presence of D-galactose, the structure of OLL has some change and become more tight.

关 键 词：雷丸凝集素 理化性质 纯化 半乳糖 

分 类 号：Q946[生物学—植物学]