BIOCHEMISTRY OF CYSTIC FIBROSIS MUCOUS SECRETIONS

过多的粘性和坚韧的存在 气管、支气管区的粘液分泌物最多 囊性纤维化的广为人知的特征。这些粘液 分泌物会导致呼吸道阻塞，并导致慢性 肺部疾病在临床上占主导地位， 决定了大多数慢性萎缩性胃炎患者的命运。目标是 这项研究的目的是探讨改变的分子基础 纤维支气管液的粘弹性特性。这个 这项提议的具体目标是a)继续孤立， 粘液糖蛋白(粘蛋白)的纯化及性质研究 从慢性支气管炎、哮喘患者的气管、支气管分泌物中， 支气管炎和正常健康人；b)调查 不同浓度的氯化钠对黄曲霉毒素的影响 分子聚集、构象(形状)和粘弹性 纯化的纤维粘蛋白和非纤维粘蛋白的性质；c)考察 释放的整体肽的生化性质 纯化的天然纤维和非纤维粘蛋白的二硫键还原； 以及d)分离和鉴定存在于土壤中的粘蛋白种类 从CF和非CF分泌物中制备纯化的粘蛋白。这个 粘蛋白将使用本文件中建立的方案进行提纯 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来分离 色谱图。分离的粘蛋白以及各种粘蛋白 来自CF和非CF分泌物的物种将被表征 它们的生化、生物物理和免疫学特性。 生化特性将包括：测定 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， 我们将研究构象和聚集性质 使用最先进的技术，如静态光散射 和荧光偏振技术。粘弹性研究 将使用磁性显微测速仪进行测量。这个 粘蛋白的蛋白质核心的结构将通过 免疫学和生化方法。这些研究将有助于 对S变更责任机制的认识 纤维支气管液的粘弹性特性。这 知识对于发展一种理性的 慢性肺病在慢性阻塞性肺疾病中的治疗探讨 最终将需要有能力控制粘度、速度 分泌物的分泌和排出。

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.