In-depth Site-specific Analysis of N-glycoproteome in Human Cerebrospinal Fluid and Glycosylation Landscape Changes in Alzheimer's Disease.
In-depth Site-specific Analysis of N-glycoproteome in Human Cerebrospinal Fluid and Glycosylation Landscape Changes in Alzheimer's Disease.
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DOI：
10.1016/j.mcpro.2021.100081
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Chen Z;Yu Q;Yu Q;Johnson J;Shipman R;Zhong X;Huang J;Asthana S;Carlsson C;Okonkwo O;Li L
脑脊液（CSF）作为与中枢神经系统（CNS）细胞外液直接交换的体液，是CNS相关疾病生物标志物发现的丰富来源。已经对CSF进行了广泛的蛋白质组分析，但缺乏旨在阐明位点特异性CSF N-糖蛋白质组的研究。CSF中位点特异性N-糖蛋白质组学研究的初步努力产生有限的覆盖范围，阻碍了CSF中基于糖基化的疾病生物标志物发现的进一步实验设计。在本研究中，我们已经开发了一种N-糖蛋白质组学的方法，结合增强的N-糖肽顺序富集的亲水相互作用色谱（HILIC）和硼酸富集与电子转移和高能碰撞解离（EThcD）的大规模完整的N-糖肽分析。所开发的方法的应用程序的人CSF样品的分析，使总共2893个完整的N-糖肽从511个N-糖位点和285个N-糖蛋白的鉴定。据我们所知，这是迄今为止报道的最大的CSF位点特异性N-糖蛋白组数据集。这些数据集为更好地理解糖蛋白的结构-功能关系及其在CNS相关生理和病理过程中的作用提供了分子基础。随着越来越多的证据表明糖基化缺陷参与阿尔茨海默病（AD）的发病机制，在本研究中，对来自健康对照和AD患者的CSF样品进行了比较深入的N-糖蛋白组学分析，其产生了相当的N-糖蛋白组覆盖率，但不同类别的糖型具有不同的表达模式，例如AD CSF样品中岩藻糖基化降低。检测到包括α-1-抗胰凝乳蛋白酶、肝配蛋白-A3和肌肽酶CN 1等在内的许多N-糖蛋白的糖基化模式改变，其作为进一步基于糖基化的AD研究的潜在感兴趣的靶点，并可能最终导致糖基化在AD进展中的作用的分子阐明。通过HILIC和硼酸富集有效地连续富集N-糖肽。使用EThcD的位点特异性完整N-糖肽表征。人CSF中的深入位点特异性N-糖蛋白质组分析。阿尔茨海默病中糖基化模式的图谱。已经进行了一项探索性的基于糖基化的生物标志物研究，以深入绘制从健康对照和阿尔茨海默病（AD）受试者的脑脊液（CSF）中收集的糖蛋白组的总体糖基化景观和位点特异性改变。比较将阐明糖蛋白质谱，主要的糖基化差异和相似性，以及一些有趣的候选糖蛋白与特定的糖基化模式改变在AD。
As the body fluid that directly interchanges with the extracellular fluid of the central nervous system (CNS), cerebrospinal fluid (CSF) serves as a rich source for CNS-related disease biomarker discovery. Extensive proteome profiling has been conducted for CSF, but studies aimed at unraveling site-specific CSF N-glycoproteome are lacking. Initial efforts into site-specific N-glycoproteomics study in CSF yield limited coverage, hindering further experimental design of glycosylation-based disease biomarker discovery in CSF. In the present study, we have developed an N-glycoproteomic approach that combines enhanced N-glycopeptide sequential enrichment by hydrophilic interaction chromatography (HILIC) and boronic acid enrichment with electron transfer and higher-energy collision dissociation (EThcD) for large-scale intact N-glycopeptide analysis. The application of the developed approach to the analyses of human CSF samples enabled identifications of a total of 2893 intact N-glycopeptides from 511 N-glycosites and 285 N-glycoproteins. To our knowledge, this is the largest site-specific N-glycoproteome dataset reported for CSF to date. Such dataset provides molecular basis for a better understanding of the structure–function relationships of glycoproteins and their roles in CNS-related physiological and pathological processes. As accumulating evidence suggests that defects in glycosylation are involved in Alzheimer's disease (AD) pathogenesis, in the present study, a comparative in-depth N-glycoproteomic analysis was conducted for CSF samples from healthy control and AD patients, which yielded a comparable N-glycoproteome coverage but a distinct expression pattern for different categories of glycoforms, such as decreased fucosylation in AD CSF samples. Altered glycosylation patterns were detected for a number of N-glycoproteins including alpha-1-antichymotrypsin, ephrin-A3 and carnosinase CN1 etc., which serve as potentially interesting targets for further glycosylation-based AD study and may eventually lead to molecular elucidation of the role of glycosylation in AD progression. Efficient N-glycopeptide sequential enrichment by HILIC and boronic acid enrichment. Site-specific intact N-glycopeptide characterization using EThcD. In-depth site-specific N-glycoproteome analysis in human CSF. Mapping the landscape of glycosylation patterns in Alzheimer's disease. An exploratory glycosylation-based biomarker study has been conducted for in-depth mapping of an overall glycosylation landscape and site-specific alteration in glycoproteome collected from cerebrospinal fluids (CSF) in healthy control and Alzheimer’s disease (AD) subjects. The comparison will shed light on the glycoproteome profile, dominant glycosylation differences and similarities, and some of the interesting glycoprotein candidates with specific glycosylation pattern alterations in AD.
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影响因子：
7.8
作者：
Chen Z;Glover MS;Li L
通讯作者：
Li L
影响因子：
16.2
作者：
Bai, Bing;Wang, Xusheng;Peng, Junmin
通讯作者：
Peng, Junmin
影响因子：
2.3
作者：
Cunningham, Robert;Wang, Jingxin;Li, Lingjun
通讯作者：
Li, Lingjun
影响因子：
4.4
作者：
Bern, Marshall W.;Kil, Yong J.
通讯作者：
Kil, Yong J.
DOI：
10.1074/mcp.m111.011601
发表时间：
2012-09
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Balog CI;Stavenhagen K;Fung WL;Koeleman CA;McDonnell LA;Verhoeven A;Mesker WE;Tollenaar RA;Deelder AM;Wuhrer M
通讯作者：
Wuhrer M