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从羊血清中分离纯化甘露聚糖结合凝集素 (MBL ) ,并研究其与单糖配体的结合活性 ,为进一步研究MBL与病原微生物的识别结合活性提供研究基础。用甘露聚糖 Sepharose 4B亲和柱行亲和层析 ,从羊血清中分离纯化出MBL分子 ,行还原性SDS PAGE测定其分子量。用VII型胶原酶鉴定其胶原样结构。用酶联凝集素试验 (ELLA)鉴定其与单糖配体的结合活性。得到纯化的羊血清MBL分子有两种单体 ,相对分子质量分别为 2 90 0 0和 330 0 0。羊血清MBL具有胶原样结构 ,可以被胶原酶消化。羊血清MBL与辣根过氧化物酶表面糖链中甘露糖的结合受N 乙酰葡糖胺、L 岩藻糖、D 氨基葡糖、N 乙酰氨基半乳糖不同程度的抑制。MBL与单糖配体的结合活性预示了其潜在的与表面富含这些糖基结构的病原微生物的识别结合能力。
Isolation and purification of mannan-binding lectin (MBL) from sheep serum and its binding activity with monosaccharide ligands provide the basis for further study on the recognition and binding activity of MBL with pathogenic microorganisms. The MBL molecules were isolated from goat serum by affinity chromatography with Sepharose 4B affinity column and the molecular weight was determined by reducing SDS PAGE. Collagen-like structure was identified with type VII collagenase. Its binding activity to monosaccharide ligands was identified by enzyme linked agglutinin assay (ELLA). The purified sheep serum MBL molecule has two kinds of monomers with relative molecular mass of 2 90 0 0 and 330 0 0, respectively. Sheep serum MBL has a collagen-like structure that can be digested by collagenase. The binding of goat serum MBL to mannose in the surface sugar chain of horseradish peroxidase was inhibited to varying degrees by N-acetylglucosamine, L-fucose, D-glucosamine and N-acetylgalactosamine. The binding activity of MBL to monosaccharide ligands predicts its potential binding ability to recognize pathogenic microorganisms with these glycosyl structures on the surface.