Alhadeff J A, Andrews-Smith G L
Biochem J. 1984 Oct 15;223(2):293-8. doi: 10.1042/bj2230293.
A competitive-binding radioimmunoassay method was employed to investigate the role of carbohydrate in antigenic determinant(s) of human liver alpha-L-fucosidase. Competition curves were used to quantify the concentrations of competitors needed to cause 30% inhibition of the precipitation of 125I-labelled alpha-L-fucosidase. The isoelectric forms of alpha-L-fucosidase, which are related by sialic acid residues, were separated preparatively and used as competitors in the radioimmunoassay. A pattern of increasing effectiveness as competitors with increasing acidity of the forms was found, suggesting that sialic acid may be involved in the antigenic determinant(s) of alpha-L-fucosidase. Specificity was exhibited when sugar and sugar derivatives were used as competitors in the radioimmunoassay: a 51-fold range of competitive ability was found, and sialic acids (N-acetylneuraminic acid and N-glycollylneuraminic acid) and colominic acid (a polymer of N-acetylneuraminic acid) were the best competitors. The results of our studies suggest that carbohydrate contributes to antigenic determinant(s) of alpha-L-fucosidase and that sialic acid is probably the major sugar involved.
采用竞争性结合放射免疫测定法来研究碳水化合物在人肝α-L-岩藻糖苷酶抗原决定簇中的作用。竞争曲线用于定量引起125I标记的α-L-岩藻糖苷酶沉淀抑制30%所需的竞争者浓度。通过唾液酸残基相关的α-L-岩藻糖苷酶的等电形式被制备性分离,并用作放射免疫测定中的竞争者。发现随着形式酸度增加,作为竞争者的有效性增加的模式,表明唾液酸可能参与α-L-岩藻糖苷酶的抗原决定簇。当糖和糖衍生物用作放射免疫测定中的竞争者时表现出特异性:发现竞争能力范围为51倍,唾液酸(N-乙酰神经氨酸和N-糖基神经氨酸)和大肠杆菌素酸(N-乙酰神经氨酸的聚合物)是最佳竞争者。我们的研究结果表明,碳水化合物有助于α-L-岩藻糖苷酶的抗原决定簇,并且唾液酸可能是主要涉及的糖。