当前位置: X-MOL 学术J. Proteome Res. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Quantitative Longitudinal Inventory of the N-Glycoproteome of Human Milk from a Single Donor Reveals the Highly Variable Repertoire and Dynamic Site-Specific Changes.
Journal of Proteome Research ( IF 4.4 ) Pub Date : 2020-03-03 , DOI: 10.1021/acs.jproteome.9b00753
Jing Zhu 1, 2, 3 , Yu-Hsien Lin 1, 2 , Kelly A Dingess 1, 2 , Marko Mank 4 , Bernd Stahl 4, 5 , Albert J R Heck 1, 2
Affiliation  

Protein N-glycosylation on human milk proteins assists in protecting an infant's health and functions among others as competitive inhibitors of pathogen binding and immunomodulators. Due to the individual uniqueness of each mother's milk and the overall complexity and temporal changes of protein N-glycosylation, analysis of the human milk N-glycoproteome requires longitudinal personalized approaches, providing protein- and N-site-specific quantitative information. Here, we describe an automated platform using hydrophilic-interaction chromatography (HILIC)-based cartridges enabling the proteome-wide monitoring of intact N-glycopeptides using just a digest of 150 μg of breast milk protein. We were able to map around 1700 glycopeptides from 110 glycoproteins covering 191 glycosites, of which 43 sites have not been previously reported with experimental evidence. We next quantified 287 of these glycopeptides originating from 50 glycoproteins using a targeted proteomics approach. Although each glycoprotein, N-glycosylation site, and attached glycan revealed distinct dynamic changes, we did observe a few general trends. For instance, fucosylation, especially terminal fucosylation, increased across the lactation period. Building on the improved glycoproteomics approach outlined above, future studies are warranted to reveal the potential impact of the observed glycosylation microheterogeneity on the healthy development of infants.

中文翻译:

来自单个供体的人乳N-糖蛋白组的纵向定量清单显示了高度可变的库和动态的特定位点变化。

人乳蛋白上的蛋白N-糖基化有助于保护婴儿的健康,并作为病原体结合和免疫调节剂的竞争性抑制剂发挥作用。由于每种母乳的独特性以及蛋白质N-糖基化的整体复杂性和时间变化,对人乳N-糖基蛋白质组的分析需要纵向的个性化方法,以提供蛋白质和N位点特异性的定量信息。在这里,我们描述了一种使用基于亲水相互作用色谱(HILIC)的药盒的自动化平台,该药盒仅使用150μg母乳蛋白质的消化物就可以对完整的N-糖肽进行蛋白质组范围的监测。我们能够从110种糖蛋白(涵盖191个糖位)绘制大约1700个糖肽,其中43个站点以前没有用实验证据进行报告。接下来,我们使用靶向蛋白质组学方法对287种源自50种糖蛋白的糖肽进行了定量。尽管每个糖蛋白,N-糖基化位点和连接的聚糖均显示出明显的动态变化,但我们确实观察到了一些总体趋势。例如,在整个泌乳期,岩藻糖基化,特别是末端岩藻糖基化增加。在上面概述的改进的糖蛋白组学方法的基础上,有必要进行进一步的研究以揭示观察到的糖基化微异质性对婴儿健康成长的潜在影响。并且连接的聚糖显示出明显的动态变化,我们确实观察到了一些总体趋势。例如,在整个泌乳期,岩藻糖基化,特别是末端岩藻糖基化增加。在上面概述的改进的糖蛋白组学方法的基础上,有必要进行进一步的研究以揭示观察到的糖基化微异质性对婴儿健康成长的潜在影响。并且连接的聚糖显示出明显的动态变化,我们确实观察到了一些总体趋势。例如,在整个泌乳期,岩藻糖基化,特别是末端岩藻糖基化增加。在上面概述的改进的糖蛋白组学方法的基础上,有必要进行进一步的研究以揭示观察到的糖基化微异质性对婴儿健康成长的潜在影响。
更新日期:2020-03-03
down
wechat
bug