Comparison of the methods for profiling glycoprotein glycans - HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study
Comparison of the methods for profiling glycoprotein glycans - HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study
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DOI：
10.1093/glycob/cwl086
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发表时间：
2007-04-01
期刊：
影响因子：
4.3
通讯作者：
Taniguchi, Naoyuki
中科院分区：
文献类型：
--
作者：
Wada, Yoshinao;Azadi, Parastoo;Taniguchi, Naoyuki
糖蛋白的质谱分析是蛋白质组学的一个新兴领域，旨在满足对分子中低聚糖成分的结构复杂性和功能的分析。考虑到最近发表的用于低聚糖分析的质谱方法的分歧，有必要建立技术标准和证明能力。在人类蛋白质组组织（HUPO）人类疾病糖组学/蛋白质组计划（HGPI）的研究中，在20个实验室对相同的转铁蛋白和免疫球蛋白- g样品进行了n-连接寡糖及其相对丰度的分析，并对色谱和质谱分析结果进行了评估。总的来说，在六个实验室中进行的基质辅助激光解吸/电离（MALDI）飞行时间质谱法对过甲基化低聚糖混合物进行了很好的定量分析，结果可以与还原性胺化衍生物的色谱分析结果相关联。对于低聚糖醛醇，石墨化碳-液相色谱(LC)/电喷雾电离（ESI）质谱在负离子模式下检测去质子化分子提供了可接受的定量。三种方法的结果差异较小。使用纳米LC/ESI质谱/质谱或MALDI质谱对色氨酸糖肽进行详细分析，证明了在这些样品中确定位点特异性或亚类特异性聚糖谱的出色能力。考虑到本研究中使用的多种质谱技术和不同方案的选择，这项多机构研究的结果表明，基于质谱的分析似乎是糖组学研究中鉴定和定量低聚糖的有效方法，并支持质谱在蛋白质组学程序中具有高灵敏度的糖肽表征的能力。
Mass spectrometry (MS) of glycoproteins is an emerging field in proteomics, poised to meet the technical demand for elucidation of the structural complexity and functions of the oligosaccharide components of molecules. Considering the divergence of the mass spectrometric methods employed for oligosaccharide analysis in recent publications, it is necessary to establish technical standards and demonstrate capabilities. In the present study of the Human Proteome Organisation (HUPO) Human Disease Glycomics/Proteome Initiative (HGPI), the same samples of transferrin and immunoglobulin-G were analyzed for N-linked oligosaccharides and their relative abundances in 20 laboratories, and the chromatographic and mass spectrometric analysis results were evaluated. In general, matrix-assisted laser desorption/ionization (MALDI) time-of-flight MS of permethylated oligosaccharide mixtures carried out in six laboratories yielded good quantitation, and the results can be correlated to those of chromatography of reductive amination derivatives. For underivatized oligosaccharide alditols, graphitized carbon-liquid chromatography (LC)/electrospray ionization (ESI) MS detecting deprotonated molecules in the negative ion mode provided acceptable quantitation. The variance of the results among these three methods was small. Detailed analyses of tryptic glycopeptides employing either nano LC/ESI MS/MS or MALDI MS demonstrated excellent capability to determine site-specific or subclass-specific glycan profiles in these samples. Taking into account the variety of MS technologies and options for distinct protocols used in this study, the results of this multi-institutional study indicate that MS-based analysis appears as the efficient method for identification and quantitation of oligosaccharides in glycomic studies and endorse the power of MS for glycopeptide characterization with high sensitivity in proteomic programs.