BIOCHEMISTRY OF CYSTIC FIBROSIS MUCUS SECRETIONS

存在过量的粘性和顽固性物质 气管支气管区域的粘液分泌最多 囊性纤维化（CF）的众所周知的特征。 这些粘液 分泌物会导致气道阻塞并导致慢性疾病 肺部疾病在临床表现中占主导地位 决定了大多数CF患者的命运。 目标 这项研究的目的是调查改变的分子基础 CF 气管支气管分泌物的粘弹性特性。 的 该提案的具体目标是 a) 继续隔离， 粘液糖蛋白（粘蛋白）的纯化和表征 来自CF、哮喘患者的气管支气管分泌物， 支气管炎和正常健康个体； b) 调查 不同氯化钠浓度对 分子聚集、构象（形状）和粘弹性 纯化的 CF 和非 CF 粘蛋白的特性； c) 调查 释放的完整多肽的生化特性 纯化的天然 CF 和非 CF 粘蛋白的二硫键还原； d) 分离和表征存在于 由 CF 和非 CF 分泌物制备的纯化粘蛋白。 的 粘蛋白将使用本文件中建立的方案进行纯化 实验室。 粘蛋白种类（来自纯化的 CF 和非 CF 粘蛋白）将通过阴离子交换和免疫亲和力分离 色谱法。 分离的粘蛋白以及各种粘蛋白 CF 和非 CF 分泌物的物种将被表征 因其生物化学、生物物理和免疫学特性。 生化表征将包括： 碳酸盐、硫酸盐、氨基酸组成、硫醇和二硫化物 粘蛋白分子的键含量。 分子大小， 将研究构象和聚集特性 使用最先进的技术，例如静态光散射 和荧光偏振技术。 粘弹性研究 将使用磁微流变仪进行。 的 粘蛋白的蛋白质核心结构将通过以下方式进行研究 免疫学和生化方法。 这些研究将有助于 对负责改变的机制的理解 CF 气管支气管分泌物的粘弹性特性。 这个 为了发展理性的知识，知识是必不可少的 CF 慢性肺病的治疗方法 最终需要能够控制粘度、速率 分泌物的分泌和排除。

The presence of excessive amounts of viscous and tenacious mucus secretions in the tracheobronchial region is the most widely known feature of cystic fibrosis (CF). These mucus secretions cause obstruction of the airways and lead to chronic pulmonary disease which dominates the clinical picture and determines the fate of the majority of CF patients. The objective of this research is to investigate the molecular basis of altered viscoelastic properties of CF tracheobronchial secretions. The specific aims of this proposal are a) to continue isolation, purification and characterization of mucus glycoproteins (mucins) from tracheobronchial secretions of patients with CF, asthma, bronchitis and from normal healthy individuals; b) to investigate the effects of different sodium chloride concentrations on the molecular aggregation, conformation (shape) and viscoelastic properties of purified CF and non-CF mucins; c) to investigate the biochemical properties of integral polypeptides released upon disulfide bond reduction of purified native CF and non-CF mucins; and d) to isolate and characterize mucin species present in the purified mucins prepared from CF and non-CF secretions. The mucins will be purified using protocols established in this laboratory. The mucin species (from purified CF and non-CF mucins) will be isolated by anion-exchange and immunoaffinity chromatographies. The isolated mucins as well as various mucin species from the CF and non-CF secretions will be characterized for their biochemical, biophysical and immunological properties. Biochemical characterization will include: determination of carbodrate, sulfate, amino acid composition, thiol and dissulfide bond content of the mucin molecules. The molecular size, conformation and aggregation properties will be investigated using state-of-the-art techniques, such as, static light scattering and fluorescence polarization techniques. The viscoelastic studies will be conducted using a magnetic microrheometer. The structure of the protein cores of the mucin will be investigated by immunological and biochemical methods. These studies will aid in the understanding of mechanism(s) responsible for altered viscoelastic properties of CF tracheobronchial secretions. This knowledge will be essential in order to develop a rational approach to the treatment of chronic lung disease in CF which will ultimately require an ability to control the viscosity, rate of secretion and elimination of the secretions.