Comparison of Methods for Profiling O-Glycosylation HUMAN PROTEOME ORGANISATION HUMAN DISEASE GLYCOMICS/PROTEOME INITIATIVE MULTI-INSTITUTIONAL STUDY OF IgA1
Comparison of Methods for Profiling O-Glycosylation HUMAN PROTEOME ORGANISATION HUMAN DISEASE GLYCOMICS/PROTEOME INITIATIVE MULTI-INSTITUTIONAL STUDY OF IgA1
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DOI：
10.1074/mcp.m900450-mcp200
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发表时间：
2010-04-01
影响因子：
7
通讯作者：
Taniguchi, Naoyuki
中科院分区：
文献类型：
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作者：
Wada, Yoshinao;Dell, Anne;Taniguchi, Naoyuki
人类蛋白质组组织人类疾病糖组学/蛋白质组计划最近协调了一项多机构研究，该研究评估了广泛用于定义糖蛋白中N-聚糖含量的方法。该研究令人信服地支持质谱法作为诊断研究发现阶段糖组学分析的首选技术。本研究报告了人类疾病糖组学/蛋白质组学倡议活动的扩展，以评估目前用于O-聚糖分析的方法。将从多发性骨髓瘤患者血清中分离的三份IgA 1样本分发给全球15个实验室进行O-糖组学分析。使用了代表当前方法的各种质谱和色谱程序。与之前的N-聚糖研究相似，结果令人信服地证实了MS在O-聚糖分析方面的卓越性能。发现两种通用策略可提供最可靠的数据，即在正离子模式下对全甲基化还原聚糖混合物进行直接MS分析，以及使用LC-MS方法在负离子模式下对天然还原聚糖进行分析。此外，质谱方法分析O-糖肽也是成功的。Molecular & Cellular Proteomics 9：719-727，2010.
The Human Proteome Organisation Human Disease Glycomics/Proteome Initiative recently coordinated a multiinstitutional study that evaluated methodologies that are widely used for defining the N-glycan content in glycoproteins. The study convincingly endorsed mass spectrometry as the technique of choice for glycomic profiling in the discovery phase of diagnostic research. The present study reports the extension of the Human Disease Glycomics/ Proteome Initiative's activities to an assessment of the methodologies currently used for O-glycan analysis. Three samples of IgA1 isolated from the serum of patients with multiple myeloma were distributed to 15 laboratories worldwide for O-glycomics analysis. A variety of mass spectrometric and chromatographic procedures representative of current methodologies were used. Similar to the previous N-glycan study, the results convincingly confirmed the pre-eminent performance of MS for O-glycan profiling. Two general strategies were found to give the most reliable data, namely direct MS analysis of mixtures of permethylated reduced glycans in the positive ion mode and analysis of native reduced glycans in the negative ion mode using LC-MS approaches. In addition, mass spectrometric methodologies to analyze O-glycopeptides were also successful. Molecular & Cellular Proteomics 9: 719-727, 2010.