Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans
Analyst ( IF 3.6 ) Pub Date : 2017-10-10 00:00:00 , DOI: 10.1039/c7an01262d Shiyue Zhou 1, 2, 3, 4 , Lucas Veillon 1, 2, 3, 4 , Xue Dong 1, 2, 3, 4 , Yifan Huang 1, 2, 3, 4 , Yehia Mechref 1, 2, 3, 4
Analyst ( IF 3.6 ) Pub Date : 2017-10-10 00:00:00 , DOI: 10.1039/c7an01262d Shiyue Zhou 1, 2, 3, 4 , Lucas Veillon 1, 2, 3, 4 , Xue Dong 1, 2, 3, 4 , Yifan Huang 1, 2, 3, 4 , Yehia Mechref 1, 2, 3, 4
Affiliation
|
Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans.
中文翻译：
LC-MS / MS分析N-聚糖的衍生化策略的直接比较
蛋白质糖基化是常见的翻译后修饰，对蛋白质折叠，寿命，构象，分布和功能有重大影响。附着在蛋白质天冬酰胺残基上的N-聚糖由于其与酶促释放的相容性而被最常研究。尽管N很容易聚糖的释放，组成和结构的复杂性，以及液相色谱质谱法（LC-MS）的电离效率差，使得定量糖组学研究成为一项重大挑战。为了克服这些挑战，在进行LC-MS分析之前，聚糖几乎总是被衍生化以赋予有利的特性，例如改善的电离效率，提高的LC分离效率以及在串联MS中产生更多信息片段。LC-MS分析聚糖有多种衍生方法，每种方法都具有不同的特性，这些特性会影响聚糖在LC色谱柱上的保留和MS分析。为了为正确选择衍生化试剂和LC色谱柱提供指导，在此，我们描述了对2-氨基苯甲酰胺，普鲁卡因酰胺，氨氧基TMT的全面评估，N-聚糖分析。在研究的衍生化策略中，RFMS为中性聚糖提供了最高的MS信号增强，而全甲基化显着增强了唾液酸化聚糖的MS强度和结构稳定性。
更新日期：2017-11-20
中文翻译：
LC-MS / MS分析N-聚糖的衍生化策略的直接比较
蛋白质糖基化是常见的翻译后修饰，对蛋白质折叠，寿命，构象，分布和功能有重大影响。附着在蛋白质天冬酰胺残基上的N-聚糖由于其与酶促释放的相容性而被最常研究。尽管N很容易聚糖的释放，组成和结构的复杂性，以及液相色谱质谱法（LC-MS）的电离效率差，使得定量糖组学研究成为一项重大挑战。为了克服这些挑战，在进行LC-MS分析之前，聚糖几乎总是被衍生化以赋予有利的特性，例如改善的电离效率，提高的LC分离效率以及在串联MS中产生更多信息片段。LC-MS分析聚糖有多种衍生方法，每种方法都具有不同的特性，这些特性会影响聚糖在LC色谱柱上的保留和MS分析。为了为正确选择衍生化试剂和LC色谱柱提供指导，在此，我们描述了对2-氨基苯甲酰胺，普鲁卡因酰胺，氨氧基TMT的全面评估，N-聚糖分析。在研究的衍生化策略中，RFMS为中性聚糖提供了最高的MS信号增强，而全甲基化显着增强了唾液酸化聚糖的MS强度和结构稳定性。
京公网安备 11010802027423号