GLYCAN PROFILES IN HUMAN MYELOID CELLS ASREGULATED BY SIALIDASE ACTIVITY

唾液酸酶活性调节的人骨髓细胞中的聚糖谱

• 批准号: 8170933
• 负责人: ROBERT SACKSTEIN
• 金额: $ 0.23万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170933
• 关键词: Adhesions; Anabolism; Biochemical; Cell Differentiation process; Cell surface; Cells; Computer Retrieval of Information on Scientific Projects Database; Dendritic Cells; Equipment; Evaluation; Funding; Gel; Glycoproteins; Glycoside Hydrolases; Golgi Apparatus; Grant; Human; Institution; Laboratories; Manuscripts; Mass Spectrum Analysis; Mediating; Membrane; Modification; Molecular; Myeloid Cells; Neuraminidase; Polysaccharides; Postdoctoral Fellow; Reporting; Research; Research Personnel; Resources; Role; Source; Structure; Surface; Training; United States National Institutes of Health; Variant; base; experience; glycosyltransferase; inhibitor/antagonist; interest; neutrophil; sialyl Lewis x

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The glycan determinant CD15 (also known as Lewis x, or Lex) is a distinguishing marker for human myeloid cells and mediates neutrophil adhesion to dendritic cells. Despite broad interest in this structure, the mechanisms underlying CD15 expression remain relatively uncharacterized. Accordingly, the Sackstein laboratory has investigated the molecular basis of increasing CD15 expression associated with human myeloid cell differentiation. Flow cytometric analysis of differentiating cells together with biochemical studies using inhibitors of glycan synthesis and of sialidases showed that increased CD15 expression is not due to de novo biosynthesis of CD15, but results predominantly from induction of a(2-3)-sialidase activity, which yields CD15 from cell-surface sialyl-CD15 (also known as sialyl-Lewis x, sLex or CD15s). This differentiation-associated conversion of surface CD15s to CD15 occurs mainly on glycoproteins. Until now, modulation of post-translational glycan modifications has been attributed solely to dynamic variations in glycosyltransferase expression. The results unveiled a new paradigm by demonstrating a critical role for post-Golgi membrane glycosidase activity in the 'biosynthesis' of a key glycan determinant. The Resource provided glycan profiles to respond to a reviewers' request during evaluation of the manuscript which reported these results. The Resource is also providing access to 2D-gel and LC/MS/MS equipment and training for postdoctoral fellow Christina Silvescu from the Sackstein laboratory who has previous experience in mass spectrometry.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 聚糖决定簇 CD15（也称为 Lewis x 或 Lex）是人类骨髓细胞的区分标记，介导中性粒细胞与树突状细胞的粘附。尽管人们对这种结构广泛感兴趣，但 CD15 表达的机制 仍然相对没有特征。因此，萨克斯坦实验室研究了与人类骨髓细胞分化相关的 CD15 表达增加的分子基础。对分化细胞的流式细胞术分析以及使用聚糖合成和唾液酸酶抑制剂的生化研究表明，CD15 表达增加并非由于 CD15 的从头生物合成，而是主要由 a(2-3)-唾液酸酶活性的诱导导致，这从细胞表面唾液酸-CD15（也称为唾液酸-Lewis x、sLex 或 CD15s）产生 CD15。这种与分化相关的表面 CD15 向 CD15 的转化主要发生在糖蛋白上。到目前为止，翻译后聚糖修饰的调节仅归因于糖基转移酶表达的动态变化。研究结果揭示了一个新的范例，证明了高尔基体后膜糖苷酶活性在关键聚糖决定簇的“生物合成”中的关键作用。该资源提供了聚糖概况，以回应审稿人在评估报告这些结果的手稿期间的要求。该资源还为萨克斯坦实验室的博士后克里斯蒂娜·西尔维斯库（Christina Silvescu）提供了使用 2D 凝胶和 LC/MS/MS 设备的机会，该研究员此前曾在质谱方面拥有丰富的经验。