COMPARISON OF CF AND NON-CF MUCUS GLYCOPROTEINS

CF 和非 CF 粘液糖蛋白的比较

• 批准号: 3343558
• 负责人: NEIL H JENTOFT
• 金额: $ 11.82万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1987-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3343558
• 关键词: analytical chemistry; bronchial mucus; carbohydrate structure; cystic fibrosis; emphysema; high performance liquid chromatography; human subject; human tissue; mass spectrometry; mucins; nuclear magnetic resonance spectroscopy; oligosaccharides; sputum

In spite of the fact that tracheobronchial mucus glycoproteins (TBMGP's) play a critical role in protecting the respiratory tract from inhaled particulate material and have been implicated in the pathophysiology of several fairly common diseases such as chronic bronchitis, asthma, and cystic fibrosis, our understanding of their chemical structure is quite rudimentary. Reports of disease related changes in the carbohydrate and sulfate composition of TBMGP's, particularly to cystic fibrosis, imply that carbohydrate structures are altered. What these changes are, how they affect the physical properties of TBMGP's, and the biological processes responsible for the changes are all unknown. The short range goal of this proposal is to determine exactly how much difference there really is between TBMGP's from normal compared to those from persons with cystic fibrosis. Glycoproteins will be obtained primarily by tracheal explaint tissue culture techniques. Reduced oligosaccharides will be released by Beta-elimination and divided into neutral, sialylated, and sulfated fractions. Each fraction will then be analyzed by one or more high pressure liquid chromatographic (HPLC) methods to give an oligosaccharide pattern. Differences will be established by comparing the pattern for qualitative and quantitative changes. Preliminary results with human TBMGP's and hog gastric mucin indicate that HPLC can resolve at least 30-40 different oligosaccharides. For scientific credibility the pattern analysis must be supplemented with compositional and structural analysis of specific oligosaccharides. Compositions will be determined by HPLC of benzoylated methyl glycosides as described in the Preliminary Results section. Structural analysis will use mass spectoscopy, NMR, and the classical techniques of carbohydrate chemistry, modified to allow use of benzoylation and HPLC for detecting reaction products. The primary question to be answered is whether disease related changes in TBMGP's reflect the disease state itself or some secondary effect of hypersecretion such as, for example, and altered ratio of TBMGP synthesis in goblet cells compared to submucosal glands.

尽管气管粘液糖蛋白（TBMGP）是事实 在保护呼吸道免受吸入的情况下发挥关键作用 颗粒物质，并与 几种相当常见的疾病，例如慢性支气管炎，哮喘和 囊性纤维化，我们对它们的化学结构的理解非常 基本。 碳水化合物和疾病与疾病相关的变化的报告 TBMGP的硫酸盐组成，尤其是囊性纤维化，这意味着 碳水化合物结构发生了变化。 这些变化是什么，它们如何 影响TBMGP的物理特性和生物过程 负责变更都是未知的。 该提案的短范围目标是确切确定多少 与那些相比，tbmgp的差异确实存在 来自具有囊性纤维化的人。 将获得糖蛋白 主要通过气管解释组织培养技术。 减少 寡糖将通过β-省释放，并分为 中性，溶解和硫酸化的部分。 每个分数将是 通过一种或多种高压液相色谱（HPLC）方法分析 给出寡糖模式。 差异将由 比较定性和定量变化的模式。 人类TBMGP和HOG胃粘蛋白的初步结果表明 HPLC可以解决至少30-40种不同的寡糖。 对于科学 信誉性模式分析必须补充成分 和特定寡糖的结构分析。 构图将是 由苯甲酰化甲基糖苷的HPLC如前所述 初步结果部分。 结构分析将使用质量 光谱，NMR和碳水化合物化学的经典技术， 修改以允许使用苯甲酰化和HPLC检测反应 产品。 要回答的主要问题是疾病与疾病有关 TBMGP反映了疾病状态本身或某些次要作用 例如，例如，例如，TBMGP合成的比例改变了 与粘膜腺相比，在杯状细胞中。