One-Step Selective Exoenzymatic Labeling (SEEL) Strategy for the Biotinylation and Identification of Glycoproteins of Living Cells.
One-Step Selective Exoenzymatic Labeling (SEEL) Strategy for the Biotinylation and Identification of Glycoproteins of Living Cells.
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DOI：
10.1021/jacs.6b04049
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发表时间：
2016-09-14
影响因子：
15
通讯作者：
Boons GJ
中科院分区：
文献类型：
--
作者：
Sun T;Yu SH;Zhao P;Meng L;Moremen KW;Wells L;Steet R;Boons GJ
可以可视化，捕获和识别健康和患病细胞中未被基因组编码的生物分子子集的技术将为揭示多种生理和疾病过程的分子机制提供独特的机会。在这里，我们描述了一种化学报告策略，利用重组糖基转移酶和生物素功能化的相应的糖核苷酸的细胞表面糖缀合物的标记。这种方法，称为一步选择性酶外标记，或SEEL，极大地提高了富集和识别大量的标记的糖蛋白的LC-MS/MS的能力。我们进一步证明，这种标记方法导致远优于上级富集和检测的糖蛋白在质膜相比，磺基-NHS激活的生物素化或两步SEEL。这种新方法将使人们有可能在生理和疾病状态的背景下，以前所未有的灵敏度来分析细胞表面糖蛋白组。
Technologies that can visualize, capture, and identify subsets of biomolecules that are not encoded by the genome in the context of healthy and diseased cells will offer unique opportunities to uncover the molecular mechanism of a multitude of physiological and disease processes. We describe here a chemical reporter strategy for labeling of cell surface glycoconjugates that takes advantage of recombinant glycosyltransferases and a corresponding sugar nucleotide functionalized by biotin. The exceptional efficiency of this method, termed one-step selective exoenzymatic labeling, or SEEL, greatly improved the ability to enrich and identify large numbers of tagged glycoproteins by LC–MS/MS. We further demonstrated that this labeling method resulted in far superior enrichment and detection of glycoproteins at the plasma membrane compared to a sulfo-NHS-activated biotinylation or two-step SEEL. This new methodology will make it possible to profile cell surface glycoproteomes with unprecedented sensitivity in the context of physiological and disease states.
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影响因子：
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作者：
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通讯作者：
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