An Isotope-Coded Photocleavable Probe for Quantitative Profiling of Protein O-GlcNAcylation.,ACS Chemical Biology

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An Isotope-Coded Photocleavable Probe for Quantitative Profiling of Protein O-GlcNAcylation.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2019-01-08 , DOI: 10.1021/acschembio.8b01052
Jingchao Li ₁ , Zhonghua Li ₁ , Xiaotao Duan ₂ , Ke Qin ₃ , Liuyi Dang ₄ , Shisheng Sun ₄ , Li Cai ₅ , Linda C Hsieh-Wilson ₆ , Liming Wu ₇ , Wen Yi _{1,

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Affiliation

O-linked N-acetylglucosamine ( O-GlcNAc) is a ubiquitous post-translational modification of proteins and is essential for cell function. Quantifying the dynamics of O-GlcNAcylation in a proteome-wide level is critical for uncovering cellular mechanisms and functional roles of O-GlcNAcylation in cells. Here, we develop an isotope-coded photocleavable probe for profiling protein O-GlcNAcylation dynamics using quantitative mass spectrometry-based proteomics. This probe enables selective tagging and isotopic labeling of O-GlcNAcylated proteins in one step from complex cellular mixtures. We demonstrate the application of the probe to quantitatively profile O-GlcNAcylation sites in 293T cells upon chemical induction of O-GlcNAc levels. We further applied the probe to quantitatively analyze the stoichiometry of O-GlcNAcylation between sorafenib-sensitive and sorafenib-resistant liver cancer cells, which lays the foundation for mechanistic investigation of O-GlcNAcylation in regulating cancer chemoresistance. Thus, this probe provides a powerful tool to profile O-GlcNAcylation dynamics in cells.

中文翻译：

同位素编码的光可裂解探针，用于蛋白质O-GlcNAcylation定量分析。

O-连接的N-乙酰氨基葡萄糖（O-GlcNAc）是蛋白质普遍存在的翻译后修饰，对于细胞功能至关重要。在蛋白质组范围内量化O-GlcNAcylation的动力学对于揭示细胞中O-GlcNAcylation的细胞机制和功能作用至关重要。在这里，我们开发了一种基于同位素质谱的蛋白质组学，用于分析蛋白质O-GlcNAcylation动力学的同位素编码的光裂解探针。该探针能够一步一步从复杂的细胞混合物中选择性标记和同位素标记O-GlcNAcylated蛋白。我们证明了在化学诱导O-GlcNAc水平后，该探针在293T细胞中定量分析O-GlcNAcylation位点的应用。我们进一步将该探针用于定量分析索拉非尼敏感性和索拉非尼耐药性肝癌细胞之间O-GlcNAcylation的化学计量，这为O-GlcNAcylation调控癌症化学耐药性的机理研究奠定了基础。因此，该探针提供了一个强大的工具来分析细胞中O-GlcNAcylation的动力学。

更新日期：2019-01-08

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