Intra- & interspecies variation in oligosaccharide by MS

内部-

• 批准号: 6579680
• 负责人: CARLITO B LEBRILLA
• 金额: $ 22.93万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6579680
• 关键词: SDS polyacrylamide gel electrophoresis; Xenopus oocyte; anions; biological models; carbohydrate structure; extracellular matrix; fertilization; glycoproteins; glycosylation; high performance liquid chromatography; mass spectrometry; method development; nuclear magnetic resonance spectroscopy; oligosaccharides; single cell analysis; species difference; sperm capacitation; vitelline membrane

DESCRIPTION (provided by applicant): Oligosaccharides are involved in a host of cellular processes including recognition, growth, and adhesion. Its importance to human health encompasses areas as diverse as reproduction, infection, and cancer. Structural diversity and heterogeneity is responsible for the incredible versatility of oligosaccharides, but these characteristics make it difficult to do analysis. Nonetheless, significant progress in determining structures has been made aided recently by mass spectrometry. Structural elucidation, however, is still only one-step in understanding structure-function relationship. The determination of oligosaccharide diversity is the next important step, but it represents a formidable challenge. It requires the combination of structural elucidation, rapid identification and the quantitation of expression levels. In this research the diversity of oligosaccharides in the extracellular matrix of the egg of Xenopus laevis will be determined. The egg of Xenopus laevis is an important model for vertebrate reproduction. The extracellular matrix is involved in a number of functions including protection and sperm capacitation. It consists of the fertilization layer, the vitelline envelope, and three layers of egg jelly. All four layers are composed of glycoproteins. The egg jelly is highly glycosylated with 60% glycans by weight. The biological functions of the layers are largely known but the specific roles of the oligosaccharides are relatively unknown. Methods for characterizing oligosaccharide diversity will be developed to provide analytical tools for glycomics. These methods will employ primarily mass spectrometry and will be used to rapidly elucidate structures of unknown neutral and anionic components, identify known components, and quantitate their abundances. The diversity of oligosaccharides between individuals and between individual eggs will be determined. In the process, methods for the oligosaccharide analysis of single eggs will be developed. Large diversity is expected between individuals to conform to the theory of exogenous pressures as the driving force for diversity. However, oligosaccharide diversity between individual eggs will be small. As part of this project, the vitelline envelope, or the fertilization layer, will be analyzed for its oligosaccharide content. The vitelline envelope is analogous to the zona pellucida in mammals. This research will lay the foundation for the analysis of single mammalian eggs.

描述（由申请人提供）：低聚糖参与许多细胞过程，包括识别、生长和粘附。 它对人类健康的重要性涵盖生殖、感染和癌症等多个领域。 结构多样性和异质性是寡糖令人难以置信的多功能性的原因，但这些特征使得分析变得困难。尽管如此，最近在质谱分析的帮助下，在确定结构方面取得了重大进展。然而，结构阐明仍然只是理解结构与功能关系的第一步。寡糖多样性的测定是下一个重要步骤，但它是一个艰巨的挑战。它需要结构阐明、快速鉴定和表达水平定量相结合。 在本研究中，将测定非洲爪蟾卵细胞外基质中寡糖的多样性。非洲爪蟾的卵是脊椎动物繁殖的重要模型。细胞外基质涉及多种功能，包括保护和精子获能。它由受精层、卵黄膜和三层卵冻组成。所有四层均由糖蛋白组成。蛋冻高度糖基化，含有按重量计 60% 的聚糖。这些层的生物学功能在很大程度上是已知的，但寡糖的具体作用相对未知。将开发表征寡糖多样性的方法，为糖组学提供分析工具。这些方法将主要采用质谱法，用于快速阐明未知中性和阴离子成分的结构，识别已知成分并定量其丰度。将确定个体之间和个体鸡蛋之间寡糖的多样性。在此过程中，将开发单个鸡蛋的寡糖分析方法。 预计个体之间存在巨大的多样性，以符合外生压力作为多样性驱动力的理论。然而，各个鸡蛋之间的寡糖多样性很小。作为该项目的一部分，将分析卵黄膜或受精层的寡糖含量。 卵黄膜类似于哺乳动物的透明带。这项研究将为分析单个哺乳动物卵奠定基础。