Glycosyltransferase activities of Ehrlich ascites tumor cells: detection, isolation, and characterization using oligosaccharide-Synsorb beads.
Glycosyltransferase activities of Ehrlich ascites tumor cells: detection, isolation, and characterization using oligosaccharide-Synsorb beads.
复制标题
艾氏腹水肿瘤细胞的糖基转移酶活性：使用寡糖-Synsorb 珠进行检测、分离和表征。
DOI：
10.1016/0003-9861(87)90109-3
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发表时间：
1987
影响因子：
3.9
通讯作者：
Goldstein,IJ
中科院分区：
文献类型：
--
作者：
Elices,MJ;Goldstein,IJ
埃利希肿瘤细胞膜的洗涤剂提取物表现出大量的糖基转移酶活性，这些活性已经用固定在Synsorb珠上的低聚糖作为受体进行了研究。糖苷酶消化结合不溶性产物的甲基化分析表明存在α(1,3)-半乳糖基转移酶和β(1,3)- n -乙酰氨基葡萄糖基转移酶，这些酶利用-乙酰乳胺作为受体底物。这两种酶可能参与了埃利希细胞表面α-d-半乳糖基端聚n -乙酰乳胺聚糖的生物合成。此外，还鉴定了一种作用于n -乙酰氨基葡萄糖的β-半乳糖转移酶和一种能够将GlcNAc结合到三糖β-d-GlcNAc(1,3)-β-d-Gal(1,4)-β-d-Glc-Synsorb中的β- n -乙酰氨基葡萄糖转移酶。用β-GlcNAc-Synsorb珠层析分离了埃利希细胞α-和β-半乳糖转移酶。在mncl2和UDP的存在下，β-半乳糖转移酶被特异性地吸附到单糖柱上，而α-半乳糖转移酶则不受阻碍地通过。
Detergent extracts of Ehrlich tumor cell membranes exhibit a host of glycosyltransferase activities which have been investigated using oligosaccharides immobilized to Synsorb beads as acceptors. Glycosidase digestions in combination with methylation analysis of the insoluble products have demonstrated the presence of an α(1,3)-galactosyltransferase and a β(1,3)-N-acetylglucosaminyltransferase, enzymes that utilizeN-acetyllactosamine as their acceptor substrate. The two enzymes are presumably involved in the biosynthesis of α-d-galactosyl-terminated poly-N-acetyllactosamine glycans that occur on the surface of Ehrlich cells. In addition, a β-galactosyltransferase acting onN-acetylglucosamine and a separate β-N-acetylglucosaminyltransferase that is capable of incorporating GlcNAc into the trisaccharide β-d-GlcNAc(1,3)-β-d-Gal(1,4)-β-d-Glc-Synsorb have been identified. The Ehrlich cell α- and β-galactosyltransferases have been separated by chromatography on β-GlcNAc-Synsorb beads. In the presence of MnCl2and UDP the β-galactosyltransferase is specifically adsorbed to the monosaccharide column whereas the α-galactosyltransferase passes through unretarded.