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A novel method to identify nuclear proteins modified with N-acetylglucosamine by using a soluble glycosyltransferase having nuclear-localization signal

A novel method to identify nuclear proteins modified with N-acetylglucosamine by using a soluble glycosyltransferase having nuclear-localization signal
利用具有核定位信号的可溶性糖基转移酶鉴定 N-乙酰氨基葡萄糖修饰的核蛋白的新方法
批准号:
24659026
负责人:
YAMAMOTO Kazuo
金额:
$2.5万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for Challenging Exploratory Research
财政年份:
2012
资助国家:
日本
项目状态:
已结题
起止时间:
2012-04-01 至 2014-03-31

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中文摘要
翻译
蛋白质的糖基化发生在内质网和高尔基体的管腔中,这导致了碳水化合物部分的不同结构。相比之下,N-乙酰氨基葡萄糖(GlcNAc)与丝氨酸和苏氨酸的加成是发生在细胞核和细胞质中的一种独特的糖基化反应。我们表达了一个在细胞内具有核定位信号的可溶性糖基转移酶,并诱导了GlcNAc残基的进一步延长。通过凝集素亲和层析和质谱分析,我们全面鉴定了O-GlcNAc修饰蛋白及其修饰的氨基酸残基。
英文摘要
Glycosylation of proteins occurs in the lumens of the endoplasmic reticulum and the Golgi apparatus, which resulted in diverse structures of carbohydrate moieties. By contrast, addition of N-acetylglucosamine (GlcNAc) to serine and threonine is a unique glycosylation reaction occurred in nucleus and cytoplasm. We expressed a soluble glycosyltransferase having nuclear-localization signal in the cell and induced further elongation of GlcNAc residue. By using lectin affinity chromatography and mass spectrometry, we comprehensively identified O-GlcNAc modified proteins and their GlcNAc-modified amino acid residues.
期刊论文(2)
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会议论文
DOI: 10.1371/journal.pone.0083886
发表时间: 2013
期刊: PloS one
影响因子: 3.7
作者: [Soga K, Teruya F, Tateno H, Hirabayashi J, Yamamoto K]
通讯作者: Yamamoto K
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海外基金