论文部分内容阅读
目的 研究水溶性和尿素溶性粉尘螨糖蛋白的组分及结合糖链的构成。 方法 用粉尘螨脱脂干粉提取水溶性抗原 (DerfA)和尿素溶性抗原 (DerfU) ,经SDS PAGE ,Westernblot ,免疫染色 ,胶体金染色和糖定性检测两种抗原糖蛋白的组分 ,并用 9种HRP 植物血凝素进行结合糖链鉴定。 结果 DerfA显示 4条蛋白带 ,分别为 18ku、3 0ku、5 0ku和5 8ku ,DerfU显示 2条蛋白带 (4 6ku、5 8ku) ,其中 5 8ku糖蛋白检测阳性。DerfA存在α 甘露糖 (α Man)、N 乙酰葡萄糖胺(GlcNAC)、α 葡萄糖 (α Glc) ,不存在寡糖 (Oligosaccharide)、岩藻糖 (α Fuc)、β 半乳糖 (β Gal)、β 半乳糖 乙酰氨基半乳糖 (β Gal GalNAC)及唾液酸 (AS) ;DerfU只存在α 甘露糖 ,不存在其它 7种糖链和唾液酸。 结论 α 甘露糖是粉尘螨优势糖蛋白 ,其次是N 乙酰葡萄糖胺和α 葡萄糖。
Aim To study the components of water-soluble and urea-soluble dust mite glycoproteins and the structure of sugar chains. Methods DerfA and DerfU were extracted from dermatophagoides farinae, and the components of two antigen glycoproteins were detected by SDS PAGE, Western blot, immunostaining, colloidal gold staining and glycometabolism. Nine kinds of HRP Phytohemagglutinin is bound to a sugar chain. Results DerfA showed four protein bands, 18ku, 30ku, 50ku and 58ku, respectively. DerfU showed two protein bands (46ku, 58ku), of which 5ku protein was positive. DerfA is present in the presence of α Man, N Acetyl Glucosamine (GlcNAC), α Glucose (α Glc), Oligosaccharide, α Fuc, β Gal, β Gal GalNAc and sialic acid (AS). DerfU only contains α-mannose and no other 7 kinds of sugar chain and sialic acid exist. Conclusion α-mannose is the dominant glycoprotein of dust mites, followed by N-acetylglucosamine and α-glucose.