Farrar G H, Uhlenbruck G, Holz G
Biochim Biophys Acta. 1980 Dec 2;603(1):185-97. doi: 10.1016/0005-2736(80)90401-0.
Affinity chromtography has been used to isolate and compare the peanut agglutinin receptors from neuraminidase-treated human, bovine and porcine erythrocyte membranes. Passage of Triton X-100-solubilised membrane material through either Sepharose- or acrylamide-based affinity columns resulted in the reversible binding of receptor molecules to the immobilised lectin. Elution with 0.2 M galactose released specifically bound glycoprotein fractions, the composition and molecular weights of which were determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. Carbohydrate analysis by gas chromatography identified these bound glycoprotein fractions as the major sources of the O-glycosidic-linked disaccharide galactosyl-beta-(1 leads to 3)-N-acetylgalactosamine in these membranes. It is suggested that these isolated fractions represent a discrete population of glycoproteins within the membranes studied, which possess both O-glycosidic- and N-glycosidic-linked carbohydrates.
亲和层析已被用于从经神经氨酸酶处理的人、牛和猪红细胞膜中分离和比较花生凝集素受体。用Triton X - 100溶解的膜材料通过基于琼脂糖或丙烯酰胺的亲和柱，导致受体分子与固定化凝集素可逆结合。用0.2M半乳糖洗脱可释放特异性结合的糖蛋白组分，其组成和分子量通过在十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳来确定。通过气相色谱进行的碳水化合物分析确定这些结合的糖蛋白组分是这些膜中O - 糖苷键连接的二糖半乳糖基-β-(1→3)-N - 乙酰半乳糖胺的主要来源。有人认为，这些分离的组分代表了所研究膜内的离散糖蛋白群体，它们同时拥有O - 糖苷键连接和N - 糖苷键连接的碳水化合物。
