首页> 外文期刊>Journal of mass spectrometry: JMS >Mass spectrometric analysis of the glycosphingolipid-derived glycans from miniature pig endothelial cells and islets: identification of NeuGc epitope in pig islets
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Mass spectrometric analysis of the glycosphingolipid-derived glycans from miniature pig endothelial cells and islets: identification of NeuGc epitope in pig islets

机译:微型猪内皮细胞和胰岛糖鞘脂来源的聚糖的质谱分析:猪胰岛中NeuGc表位的鉴定

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Glycosphingolipid (GSL) is a major component of the plasma membrane in eukaryotic cells that is involved directly in a variety of immunological events via cell-to-cell or cell-to-protein interactions. In this study, qualitative and quantitative analyses of GSL-derived glycans on endothelial cells and islets from a miniature pig were performed and their glycosylation patterns were compared. A total of 60 and 47 sialylated and neutral GSL-derived glycans from the endothelial cells and islets, respectively, were characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and collision-induced fragmentation using positive-ion electrospray ionization (ESI) ion-trap tandem mass spectrometry (MS/MS). In accordance with previous immunohistochemistry studies, the alpha-Gal-terminated GSL was not detected but NeuGc-terminated GSLs were newly detected from miniature pig islets. In addition, the neutral GSL-derived glycans were relatively quantified by derivatization with carboxymethyl trimethylammonium hydrazide (so called Girard's T reagent) and MALDI-TOF MS. The structural information of the GSL-derived glycans from pig endothelial cells and islets suggests that special attention should be paid to all types of glycoconjugates expressed on pig tissues or cells for successful clinical xenotransplantation.
机译:糖鞘脂(GSL)是真核细胞质膜的主要成分,它通过细胞间或细胞间蛋白质相互作用直接参与各种免疫学事件。在这项研究中,对小型猪的内皮细胞和胰岛上的GSL衍生的聚糖进行了定性和定量分析,并比较了它们的糖基化模式。分别通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)和碰撞诱导的裂解来表征来自内皮细胞和胰岛的分别60和47个唾液酸化和中性GSL衍生的聚糖使用阳离子电喷雾电离(ESI)离子阱串联质谱(MS / MS)。根据先前的免疫组织化学研究,未检测到以α-Gal终止的GSL,但从小型猪胰岛中新发现了以NeuGc终止的GSL。另外,通过用羧甲基三甲基酰肼铵(所谓的吉拉德T试剂)和MALDI-TOF MS衍生化,相对定量中性GSL衍生的聚糖。来自猪内皮细胞和胰岛的GSL衍生聚糖的结构信息表明,应特别注意在猪组织或细胞上表达的所有类型的糖缀合物,以成功进行临床异种移植。

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