柱状田头菇凝集素的分离纯化及其部分性质

The Isolation-Purification and Some Characteristics of Agrocybe cylindracea Lectin

经硫酸铵分级沉淀、活性炭柱吸附、DEAE SepharoseFF离子交换层析及SephadexG 100分子筛层析,从柱状田头菇子实体浸出液中分离到在PAGE上显示1条蛋白质染色带的凝集素ACL(Agrocybecylindracealectin)。经SephadexG 100测得ACL分子量为67890,经SDS PAGE测定,其亚基分子量分别为34673、20000和13183。ACL的中性糖含量为5.87%,氨基酸中天冬氨酸含量最高,半胱氨酸含量最低。ACL能凝集多种动物和人的A、B或O型红细胞,对鼠红细胞的血凝活性可被7种单糖和3种双糖抑制。ACL是一种对热较为稳定的凝集素。

Agrocybe cylindracea lectin (ACL) was isolated and purified from Agrocybe cylindracea sporophore by extraction with PBS followed by (NH_4)_2SO_4 fractionation precipitation, absorption in active carbon column, ion-exchange chromatography on DEAE-Sepharose FF and gel filtration on Sephadex G-100. The purified lectin showed a single protein band in PAGE and an molecular weight of about 67890 was measured when sephadex G-100 was used. The ACL, showing three protein bands in SDS-PAGE was composed of three kinds of subunits which had a molecular weight of 34673, 20000 and 13183 respectively. And it was proved in our determination that the ACL containing 5.87% neutral saccharide could hem agglutinate the erythrocytes of many kinds of animal blood and human blood with A, B and O blood types and its hemagglutinative activity to the mouse erythrocyte was inhibited by monosaccharide and disaccharide tested, and that the ACL was insensitive to temperature change under 50℃, while in its amino acid composition, the content of Asp was the highest and the content of Cyswas the lowest.