Hull S R, Laine R A, Kaizu T, Rodriguez I, Carraway K L
J Biol Chem. 1984 Apr 25;259(8):4866-77.
Structures of the principal O-glycosides from the major cell surface sialoglycoprotein (ASGP-1) of the MAT-B1 and MAT-C1 ascites sublines of the 13762 rat mammary adenocarcinoma have been determined. Oligosaccharitols were released by alkaline borohydride treatments of ASGP-1 and purified by gel filtration, DEAE-Sephadex ion exchange chromatography, and high performance liquid chromatography. On the basis of carbohydrate composition, methylation analysis, periodate oxidation, and exoglycosidase digestion, the five major oligosaccharides released by mild alkaline borohydride were assigned the following structures: Component II-3: (NeuAc alpha 2----3Gal beta 1----4GlcNAc beta 1----6)Ga 1 NAcOH(3----1 betaGa 1 3----2 alpha NeuAc) III-2a: (Ga 1 beta 1----4G1cNAc beta 1----6)Ga 1 NAcOH(3----1 beta Ga 1 3----2 alpha NeuAc) III-2c: (Ga 1 alpha 1----3Ga 1 beta 1----4G1cNAc beta 1----6) Ga 1 NAcOH(3----1 beta Ga 1 3----2 alpha NeuAc) IV-1a: (Ga 1 beta 1----4G 1 cNAc beta 1----6)Ga 1 NAcOH(3----1 beta Ga 1) IV-1c: (Ga 1 alpha 1----3Ga 1 beta 1----4G 1 cNAc beta 1----6) Ga 1 NAcOH(3----1 beta Ga 1) Fucosylated derivatives of III-2a, IV-1a, and IV-1c were found in smaller amounts with the fucose tentatively assigned to the 2-position of the lactosamine galactose. Components II-3, III-2a, and the fucosylated derivative of III-2A were found in both MAT-B1 and MAT-C1 sublines. The alpha-galactosides were found in detectable quantities only in subline MAT-B1. Oligosaccharides from MAT-C1 cells were enriched in sialic acid when compared to those from MAT-B1 cells. These results suggest that the 13762 ascites sublines, which bear different oligosaccharides, will provide models useful for the investigation of mechanisms regulating the expression of structures of the larger O-linked oligosaccharides.
已确定来自13762大鼠乳腺腺癌的MAT - B1和MAT - C1腹水亚系主要细胞表面唾液酸糖蛋白（ASGP - 1）的主要O - 糖苷结构。通过对ASGP - 1进行碱性硼氢化物处理释放出低聚糖醇，并通过凝胶过滤、DEAE - 葡聚糖离子交换色谱和高效液相色谱进行纯化。基于碳水化合物组成、甲基化分析、高碘酸盐氧化和外切糖苷酶消化，由温和碱性硼氢化物释放的五种主要低聚糖被赋予以下结构：组分II - 3：（NeuAcα2----3Galβ1----4GlcNAcβ1----6）GalNAcOH（3----1βGal 3----2αNeuAc）III - 2a：（Galβ1----4GlcNAcβ1----6）GalNAcOH（3----1βGal 3----2αNeuAc）III - 2c：（Galα1----3Galβ1----4GlcNAcβ1----6）GalNAcOH（3----1βGal 3----2αNeuAc）IV - 1a：（Galβ1----4GlcNAcβ1----6）GalNAcOH（3----1βGal）IV - 1c：（Galα1----3Galβ1----4GlcNAcβ1----6）GalNAcOH（3----1βGal）在III - 2a、IV - 1a和IV - 1c的岩藻糖基化衍生物中发现的岩藻糖含量较少，岩藻糖暂定为乳糖胺半乳糖的2位。在MAT - B1和MAT - C1亚系中均发现了组分II - 3、III - 2a和III - 2A的岩藻糖基化衍生物。仅在MAT - B1亚系中发现了可检测量的α - 半乳糖苷。与MAT - B1细胞的低聚糖相比，MAT - C1细胞的低聚糖富含唾液酸。这些结果表明，携带不同低聚糖的13762腹水亚系将为研究调节较大O - 连接低聚糖结构表达的机制提供有用的模型。
