Comparative glycoproteomics study on the surface of SKOV3 versus IOSE80 cell lines.
Comparative glycoproteomics study on the surface of SKOV3 versus IOSE80 cell lines.
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SKOV3 与 IOSE80 细胞系表面的比较糖蛋白组学研究
DOI：
10.3389/fchem.2022.1010642
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发表时间：
2022
影响因子：
5.5
通讯作者：
中科院分区：
文献类型：
--
作者：
目的：卵巢癌SKOV 3细胞与非癌卵巢上皮IOSE 80细胞作为对照的差异细胞表面N-糖基化的位点和结构特异性定量N-糖蛋白质组学研究。研究方法：采用C18-RPLC-MS/MS（HCD with stepped normalized collision energy）对SKOV 3和IOSE 80细胞中标记的完整N-糖肽1：1混合物进行分析，并使用位点和结构特异性完整N-糖肽搜索引擎GPSeeker对获得的原始数据集进行定性和定量搜索。结果如下：在控制光谱级错误发现率≤ 1%的情况下，从2,918个N-糖蛋白中识别出13,822个糖肽光谱匹配，具有全面的N-糖位点和N-聚糖结构信息;通过位点确定和结构诊断碎片离子分别确认了3,733个N-糖位点和3,754个N-聚糖序列结构。在三次技术重复中不少于两次观察结果的对照下，倍数变化≥1.5，p值≤ 0.05，相对于IOSE 80细胞，SKOV 3细胞中的746个DEPG被定量，其中421个上调，325个下调。结论：通过同位素标记和位点特异性和结构特异性N-糖蛋白质组学定量分析卵巢癌SKOV 3细胞的细胞表面差异N-糖基化。这项发现研究提供了假定的N-糖蛋白生物标志物候选物，用于未来使用多反应监测和生物化学方法进行验证研究。
Objective: Site- and structure-specific quantitative N-glycoproteomics study of differential cell-surface N-glycosylation of ovarian cancer SKOV3 cells with the non-cancerous ovarian epithelial IOSE80 cells as the control. Methods: C18-RPLC-MS/MS (HCD with stepped normalized collision energies) was used to analyze the 1: 1 mixture of labeled intact N-glycopeptides from SKOV3 and IOSE80 cells, and the site- and structure-specific intact N-glycopeptide search engine GPSeeker was used to conduct qualitative and quantitative search on the obtained raw datasets. Results: With the control of the spectrum-level false discovery rate ≤1%, 13,822 glycopeptide spectral matches coming from 2,918 N-glycoproteins with comprehensive N-glycosite and N-glycan structure information were identified; 3,733 N-glycosites and 3,754 N-glycan sequence structures were confirmed by site-determining and structure-diagnostic fragment ions, respectively. With the control of no less than two observations among the three technical replicates, fold change ≥1.5, and p-value ≤ 0.05, 746 DEPGs in SKOV3 cells relative to IOSE80 cells were quantified, where 421 were upregulated and 325 downregulated. Conclusion: Differential cell-surface N-glycosylation of ovarian cancer SKOV3 cells were quantitatively analyzed by isotopic labeling and site- and structure-specific N-glycoproteomics. This discovery study provides putative N-glycoprotein biomarker candidates for future validation study using multiple reaction monitoring and biochemical methods.
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影响因子：
4.4
作者：
Benicky J;Sanda M;Brnakova Kennedy Z;Grant OC;Woods RJ;Zwart A;Goldman R
通讯作者：
Goldman R
影响因子：
3.4
作者：
Abbott KL;Lim JM;Wells L;Benigno BB;McDonald JF;Pierce M
通讯作者：
Pierce M
影响因子：
2
作者：
Bast, RC;Xu, FJ;Mills, GB
通讯作者：
Mills, GB
影响因子：
50.3
作者：
Anh Tuan Nguyen;Chia, Joanne;Bard, Frederic
通讯作者：
Bard, Frederic
DOI：
10.1016/j.bbamcr.2019.03.010
发表时间：
2019-08-01
影响因子：
5.1
作者：
Ferragut, Fatima;Cagnoni, Alejandro J.;Elola, Maria T.
通讯作者：
Elola, Maria T.