泥蚶肌肉凝集素的分离纯化及部分性质  

Purification and Characterization of Muscular Lectin from Tegillarca granosa Linnaeus

作　　者：柯佳颖[1] 陈寅山[2] 戴聪杰[1] 

机构地区：[1]泉州师范学院化学与生命科学学院,福建泉州362000 [2]福建师范大学生命科学学院,福建福州350007

出　　处：《泉州师范学院学报》2008年第2期100-105,共6页Journal of Quanzhou Normal University

基　　金：福建省教育厅资助项目(JA02179)

摘　　要：泥蚶肌肉提取液经硫酸铵二步分级沉淀后，再经DEAE-Sepharose（Fast Flow）离子交换和SephacrylS-200-HR分子筛层析，从中分离纯化得到泥蚶肌肉凝集素．经测定，该凝集素分子量约为238kDa，为单个亚基的蛋白质，其相对分子量为60kDa，分子中含3．27％的糖．在氨基酸组成中，谷氨酸（Glu）含量最高，其次是丝氨酸（Ser）和天门冬氨酸（Asp）．泥蚶肌肉凝集素对人和动物红细胞均有凝集作用，其中对兔的红细跑的凝集活性最高．该凝集素对兔的红细胞的凝集活性不被测试的10种浓度为200mmol·L^-1的糖所抑制．pH6．0～7．0时具有较强活性；热稳定性较高，在30-50℃的温度范围内对兔红细胞的凝集仍保持原有活性，60℃以后活力下降；凝集活性依赖于Ca^2＋．Agglutinin with hemagglutinating activity was found in muscular of Tegillarca granosa Linnaeus. By extraction,fraction with saturated ammonium sulfate precipitation,ion exchange chromatography on the DEAE-Sepharose colum and followed by gel chromatography on the Sephacryl S-200-HR colum,an agglutinin was purified from the muscular of T. granosa. The purified lectin contains 3.27% neutral saccharide. The molecular weight is about 238 kDa,which showes 1 band on SDS-PAGE,and the relative molecular weight of these subunits were 60 kDa. In its amino acid composition,Glu has the highest content, followed by Ser and Asp. Assays for erythrocyte agglutination and sugar inhibition were done in microtiter plates. The result indicates that TGL-muscular could agglutinate erythrocytes from the natural and enzyme-treated erythrocytes of human and animals, and the highest activity was found in the agglutination with rabbit erythrocytes. The agglutinic activity of TGL-muscular was not inhibited by ten kinds of carbohydrate of 200 mmol· L^-1 The hemagglutination activity of TGL-muscular depends on Ca^2＋ ,and it shows the highest hemagglutination activity in pH 6.0-7.0. Thermostability is high,the agglutinating activities hold the line between 30-50 ℃ ,and decline beyond 60 ℃.

关 键 词：泥蚶 肌肉 凝集素 纯化 性质 

分 类 号：Q959.215.2[生物学—动物学]