BIOCHEMISTRY OF CYSTIC FIBROSIS MUCOUS SECRETIONS

存在过多的粘性和韧性 气管支气管区粘液分泌物最多 囊性纤维化（CF）的一个众所周知的特征。 这些粘液 分泌物导致气道阻塞， 肺部疾病占主导地位的临床表现， 决定了大多数CF患者的命运。 客观 这项研究的目的是研究改变的分子基础， CF气管支气管分泌物的粘弹性。 的 该建议的具体目标是a）继续隔离， 粘液糖蛋白（粘蛋白）的纯化和表征 从CF，哮喘， 支气管炎和正常健康个体; B）研究 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粘蛋白的性质; c）研究 释放的整合多肽的生物化学性质 纯化的天然CF和非CF粘蛋白的二硫键还原; 和d）分离和表征存在于所述细胞中的粘蛋白种类， 从CF和非CF分泌物制备的纯化粘蛋白。 的 粘蛋白将使用本发明中建立的方案纯化。 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分离 色谱法。 分离的粘蛋白以及各种粘蛋白 将对CF和非CF分泌物中的物种进行表征 因为它们的生物化学、生物物理学和免疫学特性。 生化表征将包括： 碳水化合物，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， 构象和聚集性质将被研究 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 将使用磁性显微流变仪进行。 的 粘蛋白的蛋白质核心的结构将通过 免疫学和生物化学方法。 这些研究将有助于 对造成改变的机制的理解 CF气管支气管分泌物的粘弹性。 这 为了形成理性，知识至关重要 治疗CF中的慢性肺病的方法， 最终将需要控制粘度、流动速率和粘度的能力。 分泌物的分泌和消除。

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.