"BRONCHIAL SECRETIONS: PHYSICAL AND CHEMICAL STUDIES"

“支气管分泌物：物理和化学研究”

• 批准号: 3344752
• 负责人: Mary C Rose
• 金额: $ 14.53万
• 依托单位: CHILDREN'S NATIONAL MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-03-01 至 1986-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3344752
• 关键词: autoradiography; bronchial mucus; carbohydrate sequence; chemical structure function; cystic fibrosis; electron microscopy; epithelium; gel; gel electrophoresis; human subject; human tissue; molecular site; mucins; nonblood rheology; oligosaccharides; radiotracer; sulfates; thiols; tissue /cell culture

Our principal objectives are to determine the chemical and physical properties of human lung mucins, to determine how perturbations of particular moities affect these physical properties, and to determine whether there are differences in the chemical and physical properties between mucins obtained from the airways of healthy individuals and from the airways of patients with chronic obstructive pulmonary diseases, such as bronchial asthma (BA), bronchitis, bronchorrhea, and cystic fibrosi (CF). In our studies on 18 samples of lung mucosal gel from a single patient with bronchial asthma, we have defined conditions for the dissolution of the gel and the purification of the mucin glycoprotein and other gel components. During the past several years we have examined the architecture of the mucin molecule--in particular the issue of carbohydraterich and naked peptide domains, and the localization of the thiol groupings implicated in the aggregation and gelation properties of human lung mucins. The analysis involves radiolabeling the thiol residues with 1-14C-iodoacetamide by reductive carboxymethylation, followed by trypsin digestion, chromatography on Sepharose 4B, and peptide mapping. The nature of the trypsin fragmentation pattern has been established and during the forthcoming grant period we will characterize several of the fragments in more detail, particularly the 68K and tr-3 fragments of the naked peptide region. In addition, we will radiolabel the hydrophobic regions of mucins via explant organ culture to aid in determining the localization of the hydrophobic residues in the glycosylated and non-glycosylated regions of the mucins. We will also use the explants to radiolabel the sugars and sulfates in the oligosaccharide chains of lung mucins. We are particularly interested in sulfate localization, since our data on BA mucin is not consistent with localization on the C-6 position of galactose, as has been reported for CF lung mucin (Roussel et al, J. Biol. Chem. 250, 2114). During the past several years we have been examining the issue of whether normal, CF, and bronchial asthmatic mucins have different compositions and structural features. Mucins from only two normals, one asthmatic, and 3 cystics have been analyzed to date. We will continue these studies using more samples and more detailed biochemical and electron microscopy studies to further compare normal and pathological lung mucins.

我们的主要目标是确定化学和物理 人肺粘蛋白的性质，以确定如何扰动 特定的部分影响这些物理性质，并确定 化学和物理性质是否存在差异 从健康个体的气道获得的粘蛋白和从 慢性阻塞性肺疾病患者的气道，如 如支气管哮喘（BA）、支气管炎、支气管炎和囊性纤维化 （CF）。 在我们的研究中，18个肺粘膜凝胶样本来自一个单一的 支气管哮喘患者，我们已经定义了 凝胶的溶解和粘蛋白糖蛋白的纯化， 其他凝胶成分。 在过去的几年里，我们审查了 粘蛋白分子的结构--特别是 富含碳水化合物和裸肽结构域，以及 硫醇基团涉及的聚集和胶凝性质， 人肺粘蛋白。 分析包括放射性标记硫醇残基 与1- 14 C-碘乙酰胺通过还原羧甲基化反应，然后 胰蛋白酶消化、Sepharose 4 B层析和肽图谱。 胰蛋白酶断裂模式的性质已经确定， 在即将到来的赠款期间，我们将描述几个 片段，特别是68 K和tr-3片段， 裸肽区。 此外，我们将放射性标记疏水 粘蛋白区域，以帮助确定 疏水残基在糖基化和 粘蛋白的非糖基化区域。 我们还将使用外植体 放射性标记肺寡糖链中的糖和硫酸盐 粘蛋白。 我们对硫酸盐定位特别感兴趣，因为我们的 关于BA粘蛋白的数据与在C-6位置上的定位不一致， 半乳糖，如已经报道的CF肺粘蛋白（Atisel等人，J. Biol. Chem. 250，2114）。 在过去的几年里，我们一直在研究 正常、CF和支气管哮喘粘蛋白是否存在差异的问题 组成和结构特征。 只有两个正常人的粘蛋白， 哮喘和3个囊肿已分析到目前为止。 我们将继续 这些研究使用更多的样本和更详细的生化和电子 显微镜研究，以进一步比较正常和病理肺粘蛋白。