Biochemical Role of Airway Mucins in Cystic Fibrosis

气道粘蛋白在囊性纤维化中的生化作用

• 批准号: 6527416
• 负责人: GOVERDHAN Pal SACHDEV
• 金额: $ 21.83万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6527416
• 关键词: Pseudomonas aeruginosa; adhesin; bacteria infection mechanism; bronchial mucus; cell adhesion; cystic fibrosis; glycosylation; human subject; mass spectrometry; molecular cloning; molecular pathology; mucins; patient oriented research; respiratory epithelium; respiratory infections; sputum

DESCRIPTION (provided by applicant): Cystic Fibrosis (CF) is the most lethal genetic disease in Caucasians and is characterized by production of excessive amounts of viscous mucus secretions in the airways of the patient. This causes airway obstruction as well as chronic bacterial infections which eventually lead to respiratory failure. Mucins provide protection to epithelia through interaction of their saccharides with bacterial adhesins. Chronic colonization with Pseudomonas aeruginosa, is considered the principal cause of death in CF patients. Our laboratory and others have shown that P. aeruginosa had considerably stronger binding affinity for CF airway mucin than normal airway mucin. These observations implicate altered glycosylation of CF mucins. Indeed, aberrant glycosylation has been reported for CF mucin. However, to date, the molecular basis of increased interaction between P. aeruginosa and CF airway mucin has not been established. We hypothesize that altered glycosylation of CF mucin is responsible for its stronger binding with P. aeruginosa. We will determine structural features of the CF mucin carbohydrate ligand(s) that provide increased binding to P. aeruginosa by preparing glycopeptides and individual saccharides from CF and control mucins. The glycopeptide(s) which show high inhibition of asialo-GM; binding to P. aeruginosa will be used to isolate 0-linked glycans for further testing of inhibitory activity and structural determination using state-of-the-art highly sensitive mass spectrometry and enzymnatic methods. Affinity gels containing selected mucin glycopeptide or mucin saccharide will be used to purify the P. aeruginosa adhesins which interact with airway mucins and glycolipids, respectively. The primary structure of the major adhesins will be determined using molecular cloning techniques. Structural characterization of major adhesins will open additional approaches to prevent the binding of P. aeruginosa to airway epithelial cells and mucins of CF patients. Information on the adhesin binding sites will permit molecular modeling, design and synthesis of potent 0-glycan inhibitors of the P. aeruginosa infection. The overall long-term goal of this study is to prevent and/or treat lung infections in CF patients.

描述（由申请人提供）：囊性纤维化（CF）是最致命的 高加索人的遗传疾病，其特征是产生过多 患者气道中的粘性粘液分泌物数量。这是原因 气道阻塞以及慢性细菌感染最终 导致呼吸衰竭。粘蛋白通过 糖的相互作用与细菌粘附素的相互作用。慢性定殖 铜绿假单胞菌被认为是CF死亡的主要原因 患者。我们的实验室和其他实验室表明，铜绿假单胞菌有 与普通气道相比，CF气道粘蛋白的结合亲和力要高得多 粘蛋白。这些观察结果暗示了CF粘蛋白的糖基化改变。的确， 据报道CF粘蛋白异常糖基化。但是，迄今为止 铜绿假单胞菌和CF气道之间相互作用增加的分子基础 粘液尚未确定。我们假设CF的糖基化改变了 粘蛋白负责其与铜绿假单胞菌更强的结合。我们将 确定CF粘蛋白碳水化合物配体的结构特征 通过制备糖肽和 来自CF和对照粘蛋白的个体糖。糖肽 表现出对Asialo-GM的高抑制作用；与铜绿假单胞菌的结合将用于 分离0连接的聚糖，以进一步测试抑制活性和 使用最先进的高敏感质量的结构测定 光谱法和酶促方法。含有选定粘蛋白的亲和力凝胶 糖肽或粘蛋白糖将用于净化铜绿假单胞菌 与气道粘蛋白和糖脂相互作用的粘附蛋白。这 主要粘附素的一级结构将使用分子确定 克隆技术。主要粘附素的结构表征将打开 防止铜绿假单胞菌与气道结合的其他方法 CF患者的上皮细胞和粘蛋白。有关粘附蛋白结合的信息 站点将允许分子建模，设计和合成有效的0-Glycan 铜绿假单胞菌感染的抑制剂。总体长期目标 研究是为了预防和/或治疗CF患者的肺部感染。