Institute of Medical Physics and Biophysics, University of Münster, Robert-Koch-Str. 31, D-48149 Münster, Germany.
Rapid Commun Mass Spectrom. 2010 Apr 15;24(7):1039-48. doi: 10.1002/rcm.4479.
Glycosphingolipids (GSLs) are major components of the outer leaflet of the cell membrane. These lipids are involved in many cell surface events and show disease-related expression changes. GSLs could thus serve as useful targets for biomarker discovery. The GSL structure is characterized by two entities: a hydrophilic glycan and a hydrophobic ceramide moiety. Both components exhibit numerous structural variations, the combination of which results in a large diversity of GSL structures that can potentially exist. Mass spectrometry (MS) is a powerful tool for high-throughput analysis of GSL expression analysis and structural elucidation. Yet, the assignment of GSL structures using MS data is tedious and demands highly specialized expertise. SysBioWare, a software platform developed for MS data evaluation in glycomics, was here applied for the MS analysis of human serum GSLs. The program was tuned to provide automated compositional assignment, supporting a variety of glycan and ceramide structures. Upon in silico fragmentation, the masses of predicted ions arising from cleavages in the glycan as well as the ceramide moiety were calculated, thus enabling structural characterization of both entities. Validation of proposed structures was achieved by matching in silico calculated fragment ions with those of experimental MS/MS data. These results indicate that SysBioWare can facilitate data interpretation and, furthermore, help the user to deal with large sets of data by supporting management of MS and non-MS data. SysBioWare has the potential to be a powerful tool for high-throughput glycosphingolipidomics in clinical applications.
糖脂（GSLs）是细胞膜外叶的主要成分。这些脂质参与许多细胞表面事件，并表现出与疾病相关的表达变化。因此，GSL 可以作为生物标志物发现的有用靶标。GSL 的结构特征由两个实体组成：亲水性聚糖和疏水性神经酰胺部分。这两个组成部分都表现出许多结构变化，它们的组合导致了大量潜在存在的 GSL 结构的多样性。质谱（MS）是一种用于高通量分析 GSL 表达分析和结构阐明的强大工具。然而，使用 MS 数据分配 GSL 结构是繁琐的，需要高度专业化的专业知识。SysBioWare 是一个为糖组学中的 MS 数据评估而开发的软件平台，在这里被应用于人类血清 GSL 的 MS 分析。该程序经过调整，可提供自动组成分配，支持多种聚糖和神经酰胺结构。通过在计算机上进行片段化，计算出聚糖和神经酰胺部分断裂产生的预测离子的质量，从而能够对这两个实体进行结构特征描述。通过将计算机模拟计算出的片段离子与实验 MS/MS 数据中的片段离子进行匹配，验证了所提出的结构。这些结果表明，SysBioWare 可以促进数据解释，并且通过支持 MS 和非 MS 数据的管理，帮助用户处理大量数据。SysBioWare 有可能成为临床应用中高通量糖脂组学的强大工具。
