Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines.
Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines.
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DOI：
10.1038/s41598-017-10714-2
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发表时间：
2017-08-31
影响因子：
4.6
通讯作者：
Cyr TD
中科院分区：
文献类型：
--
作者：
She YM;Farnsworth A;Li X;Cyr TD
2009年H1N1大流行性流感的爆发要求快速生产针对a/ California/7/2009病毒的高产疫苗，这是通过在流感蛋白血凝素和神经氨酸酶中添加或删除一个糖基化位点来实现的。在本报告中，我们系统地评估了两种高产候选重组疫苗（NIBRG-121xp和NYMC-X181A）的糖基化组成、结构分布和拓扑结构，并将各种酶解与高效液相色谱和多级质谱相结合。全长蛋白序列的蛋白质组学数据分析确定了血凝素的9个n -糖基化位点，确定了低丰度神经氨酸酶的6个n -糖基化位点和糖基结构，这些位点被高甘露糖、杂交和复杂型n -聚糖占据。共分析了约300个糖肽，并通过串联质谱手工验证。这些n -聚糖的特定结构和拓扑位置与蛋白质的空间结构和居住配体结合高度相关。有趣的是，n -聚糖的磺化、聚焦化和分割n -乙酰氨基葡萄糖也在两种流感蛋白的特定糖基化位点上被可靠地鉴定出来，这些糖基化位点可能在调节蛋白质结构和增加流感病毒重组蛋白的蛋白质丰度方面发挥关键作用。
The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. A total of ~300 glycopeptides were analyzed and manually validated by tandem mass spectrometry. The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants.
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影响因子：
7.4
作者：
Bykova, NV;Rampitsch, C;Ens, W
通讯作者：
Ens, W
影响因子：
5.4
作者：
Kordyukova, Larisa V.;Serebryakova, Marina V.;Veit, Michael
通讯作者：
Veit, Michael
DOI：
10.1042/bj20112101
发表时间：
2012-06-15
期刊：
The Biochemical journal
影响因子：
--
作者：
Jayaraman A;Koh X;Li J;Raman R;Viswanathan K;Shriver Z;Sasisekharan R
通讯作者：
Sasisekharan R
影响因子：
7.4
作者：
An, HJ;Peavy, TR;Lebrilla, CB
通讯作者：
Lebrilla, CB
影响因子：
4.4
作者：
An, Yanming;McCullers, Jonathan A.;Cipollo, John F.
通讯作者：
Cipollo, John F.