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: The University of Tokyo
• 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
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24659026/
• 关键词: 生化学; 糖鎖; 翻訳後修飾; 核内タンパク質

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.

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。

项目成果

Terminal N-acetylgalactosamine-specific leguminous lectin from Wisteria japonica as a probe for human lung squamous cell carcinoma.

• 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