Site-Specific Fucosylation Analysis Identifying Glycoproteins Associated with Aggressive Prostate Cancer Cell Lines Using Tandem Affinity Enrichments of Intact Glycopeptides Followed by Mass Spectrometry.
Site-Specific Fucosylation Analysis Identifying Glycoproteins Associated with Aggressive Prostate Cancer Cell Lines Using Tandem Affinity Enrichments of Intact Glycopeptides Followed by Mass Spectrometry.
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DOI：
10.1021/acs.analchem.7b01493
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发表时间：
2017-07-18
影响因子：
7.4
通讯作者：
Zhang H
中科院分区：
文献类型：
--
作者：
Zhou J;Yang W;Hu Y;Höti N;Liu Y;Shah P;Sun S;Clark D;Thomas S;Zhang H
糖蛋白的岩藻糖基化（Fuc）在调节蛋白质功能中起重要作用，并与包括前列腺癌（Pca）在内的几种癌症类型的发展相关。因此，近年来对Fuc糖蛋白的研究越来越受到分析领域的关注。在本文中，已经建立了一种基于完整糖肽的凝集素亲和力富集然后进行质谱分析的策略，以评估各种Fac结合凝集素对非侵袭性（NAG）和侵袭性（AG）Pca细胞系的糖位点特异性Fuc分析的特异性。评价了单独或串联使用凝集素（LCA、PSA、AAL、LTL、UEA I和AOL）和MAX提取柱的Fuc糖肽富集特异性。我们的研究结果表明，使用凝集素富集显着增加岩藻糖基化糖肽的总糖肽的比例相比，MAX富集。此外，与单独使用凝集素富集相比，串联使用凝集素然后使用MAX增加了Fuc糖肽的鉴定数量。LCA、PSA和AOL的结合能力强于AAL、LTL和UEA I。此外，LCA和PSA与核心Fuc特异性结合，而AOL、AAL和UEA I显示与核心Fuc和分支Fuc两者结合。然后应用优化的富集方法（LCA和MAX串联富集）检测两种NAG和两种AG Pca细胞系中的Fuc糖蛋白组。总共，973个完整的Fuc糖肽鉴定和定量从252 Fuc蛋白通过使用串联质量标签（TMT）标记和纳米液相色谱-质谱（nanoLC-MS/MS）分析。进一步的数据分析显示，51 Fuc糖肽在AG细胞系中的过表达超过NAG细胞的2倍。蛋白质核心岩藻糖基化的分析有很大的潜力，帮助我们了解AG Pca的侵袭活性，并可能导致AG Pca的诊断方法的发展。
Fucosylation (Fuc) of glycoproteins plays an important role in regulating protein function and has been associated with the development of several cancer types including prostate cancer (Pca). Therefore, the research of Fuc glycoproteins has attracted increasing attention recently in the analytical field. Herein, a strategy based on lectin affinity enrichments of intact glycopeptides followed by mass spectrometry has been established to evaluate the specificities of various Fuc-binding lectins for glycosite-specific Fuc analysis of nonaggressive (NAG) and aggressive (AG) Pca cell lines. The enrichment specificities of Fuc glycopeptides using lectins (LCA, PSA, AAL, LTL, UEA I, and AOL) and MAX extraction cartridges alone, or in tandem, were evaluated. Our results showed that the use of lectin enrichment significantly increased the ratio of fucosylated glycopeptides to total glycopeptides compared to MAX enrichment. Furthermore, tandem use of lectin followed by MAX increased the number of identifications of Fuc glycopeptides compared to using lectin enrichment alone. LCA, PSA, and AOL showed stronger binding capacity than AAL, LTL, and UEA I. Also, LCA and PSA bound specifically to core Fuc, whereas AOL, AAL, and UEA I showed binding to both core Fuc and branch Fuc. The optimized enrichment method with tandem enrichment of LCA followed by MAX (LCA-MAX) was then applied to examine the Fuc glycoproteomes in two NAG and two AG Pca cell lines. In total, 973 intact Fuc glycopeptides were identified and quantified from 252 Fuc proteins by using the tandem-mass-tags (TMT) labeling and nanoliquid chromatography–mass spectrometry (nanoLC–MS/MS) analysis. Further data analysis revealed that 51 Fuc glycopeptides were overexpressed more than 2-fold in AG cell lines compared to NAG cells. The analysis of protein core fucosylation has great potential for aiding our understanding of invasive activity of AG Pca and may lead to the development of diagnostic approaches for AG Pca.
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影响因子：
7.4
作者：
Toghi Eshghi S;Shah P;Yang W;Li X;Zhang H
通讯作者：
Zhang H
影响因子：
39.2
作者：
Gulati R;Gore JL;Etzioni R
通讯作者：
Etzioni R
影响因子：
4.3
作者：
do Nascimento, Antonia S. F.;Serna, Sonia;Varrot, Annabelle
通讯作者：
Varrot, Annabelle
影响因子：
4.4
作者：
Hagglund, Per;Matthiesen, Rune;Bunkenborg, Jakob
通讯作者：
Bunkenborg, Jakob
影响因子：
7
作者：
Jia, Wei;Lu, Zhuang;Qian, Xiao-Hong
通讯作者：
Qian, Xiao-Hong