Purification and Applications of a Lectin from the Mushroom Gymnopilus spectabilis
Purification and Applications of a Lectin from the Mushroom Gymnopilus spectabilis
复制标题
DOI：
10.1007/s12010-013-0665-5
复制
发表时间：
2014-02-01
影响因子：
3
通讯作者：
Franco Fraguas, Laura
中科院分区：
文献类型：
--
作者：
Albores, Silvana;Mora, Paola;Franco Fraguas, Laura
用离子交换层析法从香菇Gymnopilus spanabilis（GSL）子实体中分离得到一种凝集素。凝集素凝集小鼠红细胞，对包括N-乙酰神经氨酸在内的几种单糖表现出广泛的特异性。凝集也被抑制的糖蛋白：胎球蛋白，乳铁蛋白，重组促红细胞生成素。GSL是一种糖蛋白，含有16%的碳水化合物; SDS-PAGE显示分子量为52.1和64.4 kDa的两条带。等电聚焦显示微异质性，在pIs 5.1和5.3处有两条带。该凝集素在pH 2 ~ pH 8范围内稳定，而在pH 10时，凝集活性衰减至初始活性的50%。在40和80 ℃下孵育分别导致凝集素活性损失50%和100%。合成的GSL-琼脂糖凝胶相互作用与血清孕马促性腺激素，和至少两个亚群的这种糖蛋白被分离。转铁蛋白和可溶性GSL之间没有相互作用，而与GSL-琼脂糖凝胶实现了部分识别。末端唾液酸似乎在修饰与GSL的相互作用中起着积极的作用，这取决于凝集素是可溶的还是固定的形式。纯化的凝集素在体外对金黄色葡萄球菌和尼日尔曲霉有抑制作用。
A lectin was isolated from fruiting bodies of the mushroom Gymnopilus spectabilis (GSL) by ionic exchange chromatography. The lectin agglutinates mouse red cells exhibiting broad specificity towards several monosaccharides including the N-acetylneuraminic acid. Agglutination was also inhibited by the glycoproteins: fetuin, lactoferrin, and recombinant erythropoietin. GSL is a glycoprotein possessing 16 % of carbohydrates; the SDS-PAGE showed two bands with molecular mass of 52.1 and 64.4 kDa. Isoelectric focusing displayed microheterogeneity, with two bands at pIs 5.1 and 5.3. The lectin was stable between pH 2 and pH 8 while at pH 10, the agglutination decayed to 50% of initial activity. Incubation at 40 and 80 degrees C led to 50 and 100 % loss in activity of the lectin, respectively. Synthesized GSL-Sepharose interacts with serum pregnant mare gonadotropin, and at least two subpopulations of this glycoprotein were separated. There was no interaction between transferrin and soluble GSL while a partial recognition was achieved with GSL-Sepharose. The terminal sialic acid seems to play an active role in modifying the interaction with GSL, depending if the lectin is in a soluble or immobilized form. The purified lectin inhibited in vitro the growth of Staphylococcus aureus and Aspergillus niger.