Identification of N-Glycosites in Chicken Egg White Proteins Using an Omics Strategy
Identification of N-Glycosites in Chicken Egg White Proteins Using an Omics Strategy
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使用组学策略鉴定鸡蛋清蛋白中的 N-糖基
DOI：
10.1021/acs.jafc.7b01706
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发表时间：
2017-07-05
影响因子：
6.1
通讯作者：
Ma, Meihu
中科院分区：
文献类型：
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作者：
Geng, Fang;Wang, Jinqiu;Ma, Meihu
鸡蛋清（CEW）是天然蛋白质的完美来源，具有出色的功能特性和多种生物活性。 CEW 蛋白的糖基化结构在其功能、生物活性和过敏方面发挥着重要作用。目前的工作尝试使用组学策略来鉴定 CEW 蛋白的 N-糖基化位点。 CEW蛋白用胰蛋白酶和胰凝乳蛋白酶消化；使用 PNGase F 在 (H2O)-O-18 水中对糖肽进行富集和去糖基化，然后使用高效液相色谱/串联质谱 (HPLC-MS/MS) 进行分析。 26 个 CEW 糖蛋白中总共鉴定出了 71 个 N-糖位点。 Web-Logo 分析显示，大多数 N-糖基位点位于 N-X-T (55%) 和 N-X-S (32%)。此外，CEW clusterin的二维电泳显示了一系列在35-37 kDa水平分布的斑点，等电点范围极宽，为4.54-6.68，表明CEW clusterin糖基化的异质性。这些结果为理解CEW糖蛋白的结构、功能和生物活性提供了重要信息。
Chicken egg white (CEW) is a perfect source of natural proteins that possesses outstanding functional properties and various bioactivities. The glycosylation structure of CEW proteins plays important roles in their functions, bioactivities, and allergies. The present work attempted to identify N-glycosites of CEW proteins using an omics strategy. CEW proteins were digested with trypsin and chymotrypsin; glycopeptides were enriched and deglycosylated using PNGase F in (H2O)-O-18 water, followed by analysis using high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS). A total of 71 N-glycosites in 26 CEW glycoproteins were identified. Web-Logo analysis showed that most of the N-glycosites were at N-X-T (55%) and N-X-S (32%). Furthermore, two-dimensional electrophoresis of CEW clusterin demonstrated a series of spots horizontally distributed at 35-37 kDa with an extremely wide isoelectric point range of 4.54-6.68, indicating the heterogeneity of glycosylation of CEW clusterin. These results provided important information for the understanding of the structures, functions, and bioactivities of CEW glycoproteins.