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通过聚丙烯酰胺凝胶中的凝集素反应性估计糖蛋白碳水化合物链结构

Estimating glycoprotein carbohydrate chain structures by lectin reactivities in polyacrylamide gels.

作者信息

Nicolson G L, Irimura T

出版信息

Biol Cell. 1984;51(2):157-64. doi: 10.1111/j.1768-322x.1984.tb00294.x.

Abstract

Complex mixtures of cellular glycoproteins contain a myriad of different carbohydrate chains that cannot be easily analyzed without rigorous purification of each individual glycoprotein. We have analyzed the carbohydrate chains in complex mixtures of cellular glycoproteins by separation using sodium dodecyl-sulfate polyacrylamide gel electrophoresis and interacting the gels with several 125I-labeled lectins. By use of in situ chemical modifications of the glycoproteins after their electrophoretic separation together with the known carbohydrate-binding specifities of several lectins, it has been possible to estimate glycoprotein carbohydrate chain structures. As an example we have examined the cellular glycoproteins of a ovary-colonizing metastatic variant of B16 melanoma and report the types of carbohydrate chains that are found on various melanoma glycoproteins.

摘要

细胞糖蛋白的复杂混合物包含无数不同的糖链,若不严格纯化每种糖蛋白,就难以对其进行分析。我们通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳分离细胞糖蛋白的复杂混合物,并使凝胶与几种¹²⁵I标记的凝集素相互作用,从而分析了其中的糖链。利用糖蛋白电泳分离后的原位化学修饰以及几种凝集素已知的碳水化合物结合特异性,得以估算糖蛋白糖链结构。例如,我们研究了B16黑色素瘤卵巢转移变体的细胞糖蛋白,并报告了在各种黑色素瘤糖蛋白上发现的糖链类型。

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