De novo sequencing of heparan sulfate oligosaccharides by electron-activated dissociation.
De novo sequencing of heparan sulfate oligosaccharides by electron-activated dissociation.
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DOI：
10.1021/ac402931j
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发表时间：
2013-12-17
影响因子：
7.4
通讯作者：
Lin, Cheng
中科院分区：
文献类型：
--
作者：
Huang, Yu;Yu, Xiang;Mao, Yang;Costello, Catherine E.;Zaia, Joseph;Lin, Cheng
由于游离质子介导的大量硫酸盐损失，通过碰撞激活解离 (CAD) 来表征高度硫酸化的糖胺聚糖 (GAG) 具有挑战性。虽然可以通过使用衍生化、金属阳离子加合物和/或电喷雾增压试剂来去除游离质子，但这些步骤增加了实验工作流程的复杂性。因此，需要开发一种不需要衍生化或向电喷雾溶液中添加试剂的 GAG 测序分析方法。电子脱离解离 (EDD) 可以产生广泛且信息丰富的 GAG 碎片，而无需从前体离子中去除游离质子。然而，EDD 是一个低效的过程，通常需要消耗大量样品（通常为几微克），特别是对于高度硫酸化的 GAG 离子。在这里，我们报告说，通过改进仪器、优化电离和离子转移参数以及提高 EDD 效率，可以在液相色谱 (LC) 时间尺度上生成高度硫酸化的 GAG 的高信息 EDD 光谱，而仅消耗几纳克的样品。我们进一步表明，负电子转移解离 (NETD) 是一种更有效的 GAG 测序碎片技术，产生更少的硫酸盐损失，同时消耗更少量的样品。最后，基于高分辨率串联质谱开发了一种简单的从头 HS 测序算法。这些结果证明了 EDD 和 NETD 作为敏感分析工具的潜力，可对高度硫酸化的 GAG 进行详细、高通量、从头结构分析。
Structural characterization of highly sulfated glycosaminoglycans (GAGs) by collisionally activated dissociation (CAD) is challenging because of the extensive sulfate losses mediated by free protons. While removal of the free protons may be achieved through the use of derivatization, metal cation adducts, and/or electrospray supercharging reagents, these steps add complexity to the experimental workflow. It is therefore desirable to develop an analytical approach for GAG sequencing that does not require derivatization or addition of reagents to the electrospray solution. Electron detachment dissociation (EDD) can produce extensive and informative fragmentation for GAGs without the need to remove free protons from the precursor ions. However, EDD is an inefficient process, often requiring consumption of large sample quantities (typically several micrograms), particularly for highly sulfated GAG ions. Here, we report that with improved instrumentation, optimization of the ionization and ion transfer parameters, and enhanced EDD efficiency, it is possible to generate highly informative EDD spectra of highly sulfated GAGs on the liquid chromatography (LC) time-scale, with consumption of only a few nanograms of sample. We further show that negative electron transfer dissociation (NETD) is an even more effective fragmentation technique for GAG sequencing, producing fewer sulfate losses while consuming smaller amount of samples. Finally, a simple algorithm was developed for de novo HS sequencing based on their high resolution tandem mass spectra. These results demonstrate the potential of EDD and NETD as sensitive analytical tools for detailed, high-throughput, de novo structural analyses of highly sulfated GAGs.
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影响因子：
7
作者：
Kailemia, Muchena J.;Li, Lingyun;Amster, I. Jonathan
通讯作者：
Amster, I. Jonathan
DOI：
10.1016/j.jasms.2007.10.007
发表时间：
2008-02-01
影响因子：
3.2
作者：
Wolff, Jeremy J.;Laremore, Tatiana N.;Amster, I. Jonathan
通讯作者：
Amster, I. Jonathan
DOI：
10.1255/ejms.1120
发表时间：
2011
期刊：
European journal of mass spectrometry (Chichester, England)
影响因子：
--
作者：
Leach FE 3rd;Wolff JJ;Xiao Z;Ly M;Laremore TN;Arungundram S;Al-Mafraji K;Venot A;Boons GJ;Linhardt RJ;Amster IJ
通讯作者：
Amster IJ
DOI：
10.1016/j.jasms.2008.03.010
发表时间：
2008-06-01
影响因子：
3.2
作者：
Wolff, Jeremy J.;Laremore, Tatiana N.;Amster, I. Jonathan
通讯作者：
Amster, I. Jonathan
影响因子：
1.8
作者：
Leach, Franklin E.;Arungundram, Sailaja;Amster, I. Jonathan
通讯作者：
Amster, I. Jonathan