Thanh D B, Ly Q, Smestad Paulsen B, Wold J K
Department of Pharmacognosy, Institute of Pharmacy, University of Oslo, Norway.
Allergy. 1996 Sep;51(9):608-13. doi: 10.1111/j.1398-9995.1996.tb04680.x.
IgE-binding components in an extract of horse dander were analyzed, especially with regard to the glycoprotein allergens. After SDS-PAGE under reducing conditions and blotting, several of the glycoprotein IgE-binding components, including two distinct bands of 27 and 31 kDa, were detected. Together with several other bands, they were shown to bind to the lectins Sambucus nigra agglutinin (SNA) and Datura stramonium agglutinin (DSA), indicating terminal sialic acid linked alpha 2 --> 6 to galactose, and galactose linked beta 1 --> 4 to N-acetylglucosamine, respectively. Carbohydrate analysis after SDS-PAGE and blotting showed the presence of mannose, galactose, N-acetylglucosamine, and N-acetyl neuraminic acid in the 27-kDa band. The 14.4-kDa glycoprotein component lost its IgE-binding activity after periodate oxidation. Minor differences in allergenic activity in the other glycoprotein allergenic components were also detected, indicating that the carbohydrate part of the molecule seems to play a role in the IgE-binding activity. Two-dimensional electrophoresis and subsequent immunoblotting were performed to estimate the approximate number, molecular mass, and pI of IgE-binding components. This resulted in a cluster of such components on the blot membrane.
对马皮屑提取物中的IgE结合成分进行了分析，尤其针对糖蛋白过敏原。在还原条件下进行SDS-PAGE和印迹后，检测到了几种糖蛋白IgE结合成分，包括两条分别为27 kDa和31 kDa的不同条带。与其他几条带一起，它们被证明能与凝集素黑接骨木凝集素（SNA）和曼陀罗凝集素（DSA）结合，分别表明末端唾液酸以α2→6连接到半乳糖，以及半乳糖以β1→4连接到N-乙酰葡糖胺。SDS-PAGE和印迹后的碳水化合物分析表明，27 kDa条带中存在甘露糖、半乳糖、N-乙酰葡糖胺和N-乙酰神经氨酸。14.4 kDa的糖蛋白成分在高碘酸盐氧化后失去了其IgE结合活性。在其他糖蛋白过敏原成分中也检测到了变应原活性的微小差异，这表明分子的碳水化合物部分似乎在IgE结合活性中起作用。进行了二维电泳及随后的免疫印迹，以估计IgE结合成分的大致数量、分子量和pI。这在印迹膜上形成了这样一组成分。
