Purification, characterization and induction of a C-type lectin in the freshwater planarian Dugesia japonica

Purification, characterization and induction of a C-type lectin in the freshwater planarian Dugesia japonica
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淡水涡虫 Dugesia japonica 中 C 型凝集素的纯化、表征和诱导

DOI:
10.2478/s11535-012-0014-7
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发表时间:
2012-02
影响因子:
--
通讯作者:
Huanhuan Sun
Huanhuan Sun
中科院分区:
--
文献类型:
--
作者:
Xiufang Zhang;Lianfei Zhao;Jingjing Xie;Huanhuan Sun

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凝集素是无脊椎动物免疫防御系统的重要组成部分。鉴于凝集素的重要功能,人们对其在无脊椎动物中的特性和功能进行了大量的研究。然而,对淡水脊椎动物的凝集素研究很少,这种动物在进化上具有重要意义。在这篇文章中,我们证明了戊二醛处理的红细胞与一种对兔红细胞有选择性的凝集素的凝集作用。几种碳水化合物的血凝活性抑制试验结果表明,麦芽糖的抑制作用最强。采用直链淀粉偶联琼脂糖亲和层析法从淡水涡虫粗匀浆中纯化出一种天然凝集素。纯化后的蛋白在PAGE中为一条相对分子质量为350 kDa的条带,在SDS-PAGE中为一条约56 kDa的条带。纯化的凝集素对钙离子具有依赖性。纯化的凝集素在50℃以上的温度下活性受到抑制,最适pH为5~8。纯化的凝集素对革兰氏阴性杆菌、革兰氏阳性菌、枯草杆菌也有结合活性。此外,从损伤和细菌诱导的浮游生物中获得的纯化凝集素显示出对兔红细胞的凝集活性增强。这些结果表明,纯化的凝集素可能在淡水浮游动物的先天免疫中起重要作用。
Lectins are important components of the immune defense system of invertebrates. Given their important functions, numerous investigations have been carried out on the characterization and function of lectins in invertebrates. However, lectin studies with the freshwater planarian, an evolutionarily important animal, are rare. In this paper, we demonstrate agglutination of glutaraldehyde treated erythrocytes by a lectin with preference for rabbit erythrocytes. The result of hemagglutinating activity inhibition assays with several carbohydrates showed the most potent inhibitor was maltose. A natural lectin from the crude homogenates of freshwater planarian Dugesia japonica was purified by single step affinity chromatography using amylose-coupled agarose. The purified protein appeared as one band with a molecular mass of 350 kDa in PAGE, and as one band, approximately 56 kDa, in SDS-PAGE. The purified lectin showed dependence on calcium. The activity of the purified lectin was inhibited at temperatures greater than 50°C and showed a pH optimum between 5–8. The purified lectin also has binding activity to the Gram-negative bacteria E. coli, and the Gram-positive bacteria B. subtilis. Furthermore, the purified lectin obtained from injured and bacteria-induced planarians showed increased agglutinating activity against rabbit erythrocytes. These results suggest that the purified lectin may play an important role in the innate immunity of the freshwater planarian.
DOI: 10.1016/0145-305x(90)90005-y
发表时间: 1990-12
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