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采用改进的提取方法从糖蛋白中大规模制备PA-寡糖。

Large scale preparation of PA-oligosaccharides from glycoproteins using an improved extraction method.

作者信息

Tokugawa K, Oguri S, Takeuchi M

机构信息

Central Laboratories for Key Technology, Kirin Brewery Co., Ltd. Fukuura, Japan.

出版信息

Glycoconj J. 1996 Feb;13(1):53-6.

PMID:8785488
Abstract

We have developed a new method for the large scale preparation of pyridylaminated (PA-) oligosaccharides from glycoproteins. Phenol/chloroform extraction was adapted for the removal of protein and excess 2-aminopyridine, improving the efficiency of preparation. From a 2.5 g sample of human apo-transferrin, 25-30 mu mol of agalacto biantennary PA-oligosaccharide could be obtained. By increasing the concentration of PA-oligosaccharide substrate, we were able to detect a very low level of N-acetylglucosaminlytransferase IV activity in CHO cell extracts.

摘要

我们已经开发出一种从糖蛋白大规模制备吡啶基化（PA-）寡糖的新方法。采用苯酚/氯仿萃取法去除蛋白质和过量的2-氨基吡啶，提高了制备效率。从2.5克人载脂蛋白转铁蛋白样品中，可获得25-30微摩尔的无半乳糖双天线PA-寡糖。通过提高PA-寡糖底物的浓度，我们能够在CHO细胞提取物中检测到极低水平的N-乙酰葡糖胺基转移酶IV活性。

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本文引用的文献

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High-performance liquid chromatography of pyridylaminated saccharides.吡啶胺化糖类的高效液相色谱法
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Reexamination of the pyridylamination used for fluorescence labeling of oligosaccharides and its application to glycoproteins.用于寡糖荧光标记的吡啶胺化反应的重新审视及其在糖蛋白中的应用。
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重组鼠细胞外超氧化物歧化酶在中国仓鼠卵巢细胞中的 N-糖基化分析。
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Physiological and glycomic characterization of N-acetylglucosaminyltransferase-IVa and -IVb double deficient mice.N-乙酰氨基葡萄糖转移酶-IVa 和 -IVb 双缺失小鼠的生理学和糖组学特征。
Glycobiology. 2010 Jan;20(4):485-97. doi: 10.1093/glycob/cwp200. Epub 2009 Dec 16.
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Structure determination of a sulfated N-glycans, candidate for a precursor of the selectin ligand in bovine lung.硫酸化N-聚糖的结构测定，牛肺中选择素配体前体的候选物。
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Kinetic properties and substrate specificities of two recombinant human N-acetylglucosaminyltransferase-IV isozymes.两种重组人N-乙酰葡糖胺基转移酶-IV同工酶的动力学特性和底物特异性
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Abnormal biantennary sugar chains are expressed in human chorionic gonadotropin produced in the choriocarcinoma cell line, JEG-3.异常的双触角糖链在绒毛膜癌细胞系JEG-3产生的人绒毛膜促性腺激素中表达。
Glycoconj J. 2004;20(7-8):473-81. doi: 10.1023/B:GLYC.0000038293.37376.9f.
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Electrospray mass spectrometry and fragmentation of N-linked carbohydrates derivatized at the reducing terminus.还原端衍生化的 N-连接碳水化合物的电喷雾质谱分析与裂解
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Biochem J. 1998 May 1;331 ( Pt 3)(Pt 3):733-42. doi: 10.1042/bj3310733.
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Ectopic expression of N-acetylglucosaminyltransferase III in transgenic hepatocytes disrupts apolipoprotein B secretion and induces aberrant cellular morphology with lipid storage.N-乙酰葡糖胺基转移酶III在转基因肝细胞中的异位表达会破坏载脂蛋白B的分泌，并诱导出现伴有脂质蓄积的异常细胞形态。
Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2526-30. doi: 10.1073/pnas.95.5.2526.
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Structure determination by 1H NMR spectroscopy of (sulfated) sialylated N-linked carbohydrate chains released from porcine thyroglobulin by peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase-F.通过1H核磁共振光谱法对经肽-N4-（N-乙酰-β-氨基葡萄糖基）天冬酰胺酶-F从猪甲状腺球蛋白释放的（硫酸化）唾液酸化N-连接碳水化合物链进行结构测定。
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