Glycosylation and processing of high-mannose oligosaccharides of thyroid-stimulating hormone subunits: comparison to nonsecretory cell glycoproteins.

Glycosylation and processing of high-mannose oligosaccharides of thyroid-stimulating hormone subunits: comparison to nonsecretory cell glycoproteins.
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促甲状腺激素亚基的高甘露糖寡糖的糖基化和加工:与非分泌细胞糖蛋白的比较。

DOI:
10.1021/bi00315a001
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发表时间:
1984
期刊:
影响因子:
2.9
通讯作者:
B. Weintraub
B. Weintraub
中科院分区:
生物学3区
文献类型:
--
作者:
C. Ronin;B. Stannard;I. Rosenbloom;J. Magner;B. Weintraub

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将促甲状腺激素 (TSH) 亚基糖基化与小鼠促甲状腺肿瘤中总细胞糖蛋白的糖基化进行比较。在脉冲和脉冲追踪实验中,脂联寡糖、总细胞糖蛋白和 TSH 亚基用 [3H] 甘露糖、[3H] 半乳糖或 [3H] 葡萄糖标记。分别通过脂质提取和弱酸水解、选择性免疫沉淀或酸沉淀后胰蛋白酶和糖苷内切酶 H 处理来分离各种寡糖。寡糖的性质通过它们在纸色谱中的迁移、不同前体的相对掺入以及它们对α-甘露糖苷酶的抗性来评估。 60 分钟时,发现脂质连接寡糖由 Glc3-2Man9GlcNAc2、Man9-8GlcNAc2 和 Man5GlcNAc2 组成。标记 10 或 60 分钟时,总细胞蛋白含有 Glc3Man9GlcNAc2、Glc1Man9GlcNAc2、Man9GlcNAc2、Glc1Man8GlcNAc2、Man8GlcNAc2 和 Man7GlcNAc2。最大的寡糖 Glc3Man9GlcNAc2 的半衰期异常长,约为 2 小时。相反,在通过免疫沉淀或通过十二烷基硫酸钠凝胶电泳分离的TSH + α亚基或游离β亚基上未发现Glc3Man9GlcNAc2。相反,在 10 分钟脉冲后,在 TSH + α 亚基和 β 亚基上主要发现了 Man9GlcNAc2。当脉冲跟踪长达 2 小时时,TSH + α 亚基碳水化合物链上的 Man8GlcNAc2 逐渐增加,其含量高于 β 亚基。(摘要截断为 250 字)
Thyroid-stimulating hormone (TSH) subunit glycosylation was compared to that of total cell glycoproteins in mouse thyrotropic tumors. Lipid-linked oligosaccharides, total cell glycoproteins, and TSH subunits were labeled with either [3H]mannose, [3H]galactose, or [3H]glucose in pulse and pulse-chase experiments. The various oligosaccharides were isolated respectively by lipid extraction and mild acid hydrolysis, by selective immunoprecipitation, or by acid precipitation followed by trypsin and endoglycosidase H treatment. The nature of the oligosaccharides was assessed by their migration in paper chromatography, their relative incorporation of different precursors, and also their resistance to alpha-mannosidase. At 60 min, lipid-linked oligosaccharides were found to be composed of Glc3-2Man9GlcNAc2, Man9-8GlcNAc2, and Man5GlcNAc2. At 10 or 60 min of labeling, total cell proteins contained Glc3Man9GlcNAc2, Glc1Man9GlcNAc2, Man9GlcNAc2, Glc1Man8GlcNAc2, Man8GlcNAc2, and Man7GlcNAc2. The largest oligosaccharide, Glc3Man9GlcNAc2, had an unusually long half-life of about 2 h. In contrast, no Glc3Man9GlcNAc2 was found either on TSH + alpha subunits or on free beta subunits isolated either by immunoprecipitation or by sodium dodecyl sulfate gel electrophoresis. Instead, primarily Man9GlcNAc2 was found after a 10-min pulse both on TSH + alpha subunits and on beta subunits. When the pulse was followed by a chase up to 2 h, there was a progressive increase in Man8GlcNAc2 in higher amounts on TSH + alpha-subunit carbohydrate chains than on beta subunits.(ABSTRACT TRUNCATED AT 250 WORDS)
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发表时间: 1982-12
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发表时间: 1982
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发表时间: 1983
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DOI: --
发表时间: 1981
期刊: The Journal of biological chemistry
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发表时间: 1981
期刊: The Journal of biological chemistry
影响因子: --
作者:
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