Global Identification of O-GlcNAc-Modified Proteins

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• 作者：Animesh Nandi ; Robert Sprung ; Deb K. Barma ; Yingxin Zhao ; Sung Chan Kim ; John R. Falck ; and Yingming Zhao
• 刊名：Analytical Chemistry
• 出版年：2006
• 出版时间：January 15, 2006
• 年：2006
• 卷：78
• 期：2
• 页码：452 - 458
• 全文大小：309K
• 年卷期：v.78,no.2(January 15, 2006)
• ISSN：1520-6882

文摘

The O-linked N-acetylglucosamine (O-GlcNAc) modification of serine/threonine residues is an abundant posttranslational modification present in cytosolic and nuclearproteins. The functions and subproteome of O-GlcNAcmodification remain largely undefined. Here we report theapplication of the tagging-via-substrate (TAS) approach forglobal identification of O-GlcNAc-modified proteins. TheTAS method utilizes an O-GlcNAc azide analogue formetabolic labeling of O-GlcNAc-modified proteins, whichcan be chemoselectively conjugated for detection andenrichment of the proteins for proteomics studies. Ourstudy led to the identification of 199 putative O-GlcNAc-modified proteins from HeLa cells, among which 23 wereconfirmed using reciprocal immunoprecipitation. Functional classification shows that proteins with diversefunctions are modified by O-GlcNAc, implying that O-GlcNAc might be involved in the regulation of multiplecellular pathways.