Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides.
Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides.
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通过固相提取N连接的聚糖和含糖材料的肽对蛋白质糖基化的全面分析。
DOI：
10.1038/nbt.3403
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发表时间：
2016-01
影响因子：
46.9
通讯作者：
Zhang H
中科院分区：
文献类型：
--
作者：
Sun S;Shah P;Eshghi ST;Yang W;Trikannad N;Yang S;Chen L;Aiyetan P;Höti N;Zhang Z;Chan DW;Zhang H
蛋白质糖基化的全面表征对于理解糖蛋白的结构和功能至关重要。然而，由于糖蛋白构象的复杂性和异质性，目前的糖蛋白分析主要集中在去糖基化的糖基化位点（糖基）-含有肽或释放的聚糖。在这里，我们描述了一种化学酶的方法，称为固相萃取的N-连接的聚糖和含糖基肽（NGAG）的糖蛋白的全面表征，能够确定聚糖的异质性，除了提供有关的总N-连接的聚糖，含糖基肽和糖蛋白的复杂样品的内容。NGAG方法也可用于定量检测总和位点特异性聚糖结合蛋白中的糖蛋白变化。
Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies.