Identification and Structural Characterization of Novel Chondroitin/Dermatan Sulfate Hexassacharide Domains in Human Decorin by Ion Mobility Tandem Mass Spectrometry.
Identification and Structural Characterization of Novel Chondroitin/Dermatan Sulfate Hexassacharide Domains in Human Decorin by Ion Mobility Tandem Mass Spectrometry.
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DOI：
10.3390/molecules27186026
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发表时间：
2022-09-15
期刊：
影响因子：
--
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硫酸软骨素(CS)和硫酸皮肤素(DS)在自然界中被发现与蛋白多糖相连，最常见的是CS/DS杂合链。在高表达CS/DS的细胞外基质中，CS/DS参与基本过程和各种病理过程。CS/DS结构域的结构多样性促使人们努力开发有效的分析方法，其中质谱仪(MS)是鉴定新物种和阐明其结构的最有用的技术之一。在此背景下，我们介绍了一种基于离子迁移率分离MS和碰撞诱导解离MS/MS的快速、灵敏和可靠的方法，用于分析人胚胎肾脏HEK293细胞核心蛋白中的CS/DS六糖结构域，该蛋白是通过β消除释放CS/DS链、软骨素AC裂解酶解聚和尺寸排除层析分离得到的。通过IMS MS，我们发现了新的CS/DS物种，即在释放的CS/DS链中存在过低和过硫酸盐化的六糖结构域。在最后的分析阶段，优化的IMS CID MS/MS提供了一系列诊断碎片离子，这些碎片离子对于表征三硫酸盐-4，5-Δ-GlcAGalNAc[IdoAGalNAc]2序列的硫化密码子中由于不寻常的硫化位点而发生的错误调控至关重要。
Chondroitin sulfate (CS) and dermatan sulfate (DS) are found in nature linked to proteoglycans, most often as hybrid CS/DS chains. In the extracellular matrix, where they are highly expressed, CS/DS are involved in fundamental processes and various pathologies. The structural diversity of CS/DS domains gave rise to efforts for the development of efficient analytical methods, among which is mass spectrometry (MS), one of the most resourceful techniques for the identification of novel species and their structure elucidation. In this context, we report here on the introduction of a fast, sensitive, and reliable approach based on ion mobility separation (IMS) MS and MS/MS by collision-induced dissociation (CID), for the profiling and structural analysis of CS/DS hexasaccharide domains in human embryonic kidney HEK293 cells decorin (DCN), obtained after CS/DS chain releasing by β-elimination, depolymerization using chondroitin AC I lyase, and fractionation by size-exclusion chromatography. By IMS MS, we were able to find novel CS/DS species, i.e., under- and oversulfated hexasaccharide domains in the released CS/DS chain. In the last stage of analysis, the optimized IMS CID MS/MS provided a series of diagnostic fragment ions crucial for the characterization of the misregulations, which occurred in the sulfation code of the trisulfated-4,5-Δ-GlcAGalNAc[IdoAGalNAc]2 sequence, due to the unusual sulfation sites.
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影响因子：
7.4
作者：
Gill, Vanessa Leah;Aich, Udayanath;Rao, Srinivasa;Pohl, Chris;Zaia, Joseph
通讯作者：
Zaia, Joseph
影响因子：
4.2
作者：
Sarbu, Mirela;Vukelic, Zeljka;Zamfir, Alina D.
通讯作者：
Zamfir, Alina D.
DOI：
10.1007/978-3-030-70115-4_4
发表时间：
2021-01-01
期刊：
ROLE OF GLYCOSYLATION IN HEALTH AND DISEASE
影响因子：
--
作者：
Song, Yuefan;Zhang, Fuming;Linhardt, Robert J.
通讯作者：
Linhardt, Robert J.
影响因子：
4.3
作者：
Bilong, Melanie;Bayat, Parisa;Daniel, Regis
通讯作者：
Daniel, Regis
影响因子：
4.3
作者：
Dufresne, Martin;Guneysu, Daniel;Chaurand, Pierre
通讯作者：
Chaurand, Pierre