Evaluation of Different N-Glycopeptide Enrichment Methods for N-Glycosylation Sites Mapping in Mouse Brain

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• 作者：Chengqian Zhang ; Zilu Ye ; Peng Xue ; Qingbo Shu ; Yue Zhou ; Yanlong Ji ; Ying Fu ; Jifeng Wang ; Fuquan Yang
• 刊名：Journal of Proteome Research
• 出版年：2016
• 出版时间：September 2, 2016
• 年：2016
• 卷：15
• 期：9
• 页码：2960-2968
• 全文大小：476K
• 年卷期：0
• ISSN：1535-3907

文摘

N-Glycosylation of proteins plays a critical role in many biological pathways. Because highly heterogeneous N-glycopeptides are present in biological sources, the enrichment procedure is a crucial step for mass spectrometry analysis. Five enrichment methods, including IP-ZIC-HILIC, hydrazide chemistry, lectin affinity, ZIC-HILIC-FA, and TiO₂ affinity were evaluated and compared in the study of mapping N-glycosylation sites in mouse brain. On the basis of our results, the identified N-glycosylation sites were 1891, 1241, 891, 869, and 710 and the FDR values were 3.29, 5.62, 9.54, 9.54, and 20.02%, respectively. Therefore, IP-ZIC-HILIC enrichment method displayed the highest sensitivity and specificity. In this work, we identified a total of 3446 unique glycosylation sites conforming to the N-glycosylation consensus motif (N-X-T/S/C; X ≠ P) with ¹⁸O labeling in 1597 N-glycoproteins. N-glycosylation site information was used to confirm or correct the transmembrane topology of the 57 novel transmembrane N-glycoproteins.