The distribution of repeating [Gal beta 1,4GlcNAc beta 1,3] sequences in asparagine-linked oligosaccharides of the mouse lymphoma cell lines BW5147 and PHAR 2.1.
The distribution of repeating [Gal beta 1,4GlcNAc beta 1,3] sequences in asparagine-linked oligosaccharides of the mouse lymphoma cell lines BW5147 and PHAR 2.1.
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小鼠淋巴瘤细胞系 BW5147 和 PHAR 2.1 的天冬酰胺连接寡糖中重复 [Gal beta 1,4GlcNAc beta 1,3] 序列的分布。
DOI：
10.1016/s0021-9258(20)82134-6
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Stuart Kornfeld
中科院分区：
文献类型：
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作者：
R D Cummings;Stuart Kornfeld
研究了小鼠淋巴瘤BW5147细胞中不同类型Asn-linked寡糖中重复双糖[Gal β 1,4 glcnac β 1,3]的出现和分布。从含有[6-3H]半乳糖培养基中培养的细胞中制备糖肽，并通过在魔豆蛋白A-Sepharose、豌豆凝集素- sepharose、白细胞凝集植物血凝素-琼脂糖和曼陀罗凝集素-琼脂糖上的串联凝集素亲和层析对双、三、四天线Asn-linked寡糖进行分离。如本报告所述，后一种凝集素结合的糖肽含有重复的n -乙酰乳胺序列或在C-2和C-6被n -乙酰乳胺取代的外部甘露糖残基。对分离的糖肽进行甲基化分析、特异性外糖苷酶处理和弗氏埃希菌内切- β -半乳糖苷酶消化。我们的数据表明，大约三分之二的四触角链和一半的三触角链asn寡糖在至少一个分支中含有重复的n-乙酰乳胺序列。许多重复序列包含一个额外的半乳糖残基连接α 1,3到倒数第二个半乳糖残基。相比之下，只有不到10%的双天线低聚糖含有重复双糖。在缺乏UDP-GlcNAc: α -甘露糖苷- 1,6- n -乙酰氨基葡萄糖转移酶的细胞系（PHAR 2.1）中也检测了重复n -乙酰乳胺单元的分布。这些细胞不能合成四天线和某些三天线物种，而是积累双天线低聚糖。重复n-乙酰乳胺单元的总含量在这条线上大大减少，而这些主要存在于三天线asn连接的低聚糖中。这些结果表明，重复的n-乙酰乳胺序列通常出现在BW5147细胞中复杂型Asn-linked寡糖中，但主要局限于三链和四链物种。
The occurrence and distribution of the repeating disaccharide [Gal beta 1,4GlcNAc beta 1,3] in the different types of Asn-linked oligosaccharides in mouse lymphoma BW5147 cells have been studied. Glycopeptides were prepared from cells grown in medium containing [6-3H]galactose, and the bi-, tri-, and tetraantennary Asn-linked oligosaccharides were fractionated by serial lectin affinity chromatography on concanavalin A-Sepharose, pea lectin -Sepharose, leukoagglutinating phytohemagglutinin-agarose, and Datura stramonium agglutinin-agarose. As described in this report, the latter lectin binds glycopeptides that contain either the repeating N-acetyllactosamine sequence or an outer mannose residue substituted at C-2 and C-6 by N-acetyllactosamine. The isolated glycopeptides were subjected to methylation analysis, specific exoglycosidase treatments, and digestion with Escherichia freundii endo-beta-galactosidase. Our data indicate that approximately two-thirds of the tetraantennary and one-half of the triantennary Asn-linked oligosaccharides contain repeating N-acetyllactosamine sequences in at least one branch. Many of the repeating sequences contain an additional galactose residue linked alpha 1,3 to a penultimate galactose residue. By contrast, less than 10% of the biantennary oligosaccharides contain the repeating disaccharide. The distribution of the repeating N-acetyllactosamine unit was also examined in a cell line ( PHAR 2.1) that is deficient in UDP-GlcNAc:alpha-mannoside beta 1,6-N-acetylglucosaminyltransferase. These cells are unable to synthesize tetraantennary and certain triantennary species and instead accumulate biantennary oligosaccharides. The total content of repeating N-acetyllactosamine units is greatly decreased in this line, and those that are present are found predominantly in triantennary Asn-linked oligosaccharides. These results demonstrate that the repeating N-acetyllactosamine sequence occurs commonly in complex-type Asn-linked oligosaccharides in BW5147 cells but is confined primarily to tri- and teraantennary species.