团头鲂血清抗病毒半乳糖凝集素样蛋白的分离与鉴定
Isolation and characterization of an anti-viral galectin-like protein from Megalobrama amblycephala serum
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摘要: 将草鱼出血病病毒(GCHV)的结构蛋白与Sepherose-4B偶联制备亲和柱,经亲和层析从团头鲂血清中分离到一种蛋白,能凝集兔红细胞,其血凝性依赖于β巯基乙醇的存在.红细胞凝集抑制实验证实该蛋白与D半乳糖有最高的亲和力,其次为甘露糖.梯度PAGE测得其分子量约为240 000,SDS梯度PAGE证实该蛋白由分子量为15 000的单个亚基组成,亚基之间没有共价连接.以上特性与半乳糖凝集素相符,故称其为团头鲂抗病毒半乳糖凝集素样蛋白.抗体阻断试验证实,此种凝集素样蛋白是团头鲂血清抗病毒及红细胞凝集活性的主要成分.N末段的氨基酸序列为Lys-Val-Asn-Leu-Asp-Glu-Lys-Cys-Pro-Phe,检索GenBank,未发现与这一段序列同源的蛋白质.Abstract: An anti-viral protein was isolated from Megalobrama amblycephala serum by affinity chromatography on a column of Sepharose 4B-GCHV structure protein .Depending on exogenous β-mercaptanethanol , the isolated anti-viral protein could agglutinate rabbit erythrocytes treated by trypsin .Galactose was the most active inhibitor to the protein in hemagglutination inhibition test , and mannose had less activity than that of galactose .The molecular weight of the protein was 240 000 in PG-PAGE and SDS-PG-PAGE demonstrated that it had a subunit structure consisting of a single component with a molecular weight of 15 000 .The anti-viral protein was designated anti-viral galectin-like protein of M .amblycephala .After treatment with rabbit anti-serum, the rest serum of M .amblycephala lost it' s anti-viral and hemagglutination activity completely .The amino acid sequence of N-end of the galectin-like protein was Lys-Val-Asn-Leu-Asp-Glu-Lys-Cys-Pro-Phe , and no protein with the homologous amino acid sequence was found in the GenBank .
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