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

Research Project

Project/Area Number	24659026
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Single-year Grants
Research Field	Biological pharmacy
Research Institution	The University of Tokyo
Principal Investigator	YAMAMOTO Kazuo 東京大学, 新領域創成科学研究科, 教授 (20174782)
Project Period (FY)	2012-04-01 – 2014-03-31
Project Status	Completed (Fiscal Year 2013)
Budget Amount *help	¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000) Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000) Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords	生化学 / 糖鎖 / 翻訳後修飾 / 核内タンパク質
Research Abstract	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.