The role of mucin-protein interactions in the innate defense of the lung

粘蛋白-蛋白质相互作用在肺先天防御中的作用

• 批准号: 8463602
• 负责人: Mehmet Kesimer
• 金额: $ 34.87万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-21 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8463602
• 关键词: Affinity; Antioxidants; Asthma; Behavior; Binding; Binding Proteins; Biochemical; Biological Assay; C-terminal; Cell Culture Techniques; Chemicals; Chimeric Proteins; Chronic Bronchitis; Chronic lung disease; Complex; Coughing; Coupled; Cystic Fibrosis; DMBT1 gene; Data; Defensins; Ensure; Epithelial; Epithelial Cells; Excision; Fluorescein-5-isothiocyanate; Fluorescence Recovery After Photobleaching; Future; Gel; Glycopeptides; Glycoproteins; Goals; Gold Colloid; Heart; Human; Immune system; Immunoelectron Microscopy; Infection; Inflammation; Knowledge; Label; Lead; Lung; MUC5AC gene; MUC5B gene; Mass Spectrum Analysis; Measurement; Measures; Mechanics; Methods; Microspheres; Molecular; Molecular Weight; Molecular and Cellular Biology; Morbidity - disease rate; Mucins; Mucous body substance; Predisposition; Property; Protein Binding; Protein Region; Protein-Carbohydrate Interaction; Proteins; Proteomics; Quartz; Refractive Indices; Retinal Cone; Rheology; Role; Structure; Surface; Tertiary Protein Structure; Testing; Therapeutic Intervention; Tracheobronchial; Western Blotting; antimicrobial; base; design; disease characteristic; globular protein; liquid chromatography mass spectrometry; mortality; novel; protein complex; protein structure; public health relevance; research study; respiratory

DESCRIPTION (provided by applicant): The airway epithelial mucosal barrier is the heart of a powerful innate immune system that maintains the underlying epithelial surface. At the broadest level, the major scientific goal of this proposal is to elucidate the organization of the airway gel layer, which is formed by very large glycoproteins termed mucins (MUC5B and MUC5AC) coupled with other protective biomolecules through labile interactions, and assess the role of these interactions in airway mucus integrity as an essential pre-requisite for therapeutic intervention. Our hypothesis is that normal airway mucus is highly structured, containing an excess of 100 proteins that form distinct protein complexes many of which are centered around these mucins. These complexes constitute a discrete secretory entity we call the 'mucin interactome'. We propose that gel properties emerge from a dynamic interplay between the mucins and proteins and that these structures engender specific rheological properties of mucus that tune it to removal by cough and flow clearance. This proposal will change the paradigm that mucins are the only molecules that give most of the rheological properties to mucus and will ultimately lead to a greater understanding of the role of these interactions as an innate defense of the lung. In testing these hypotheses, using a broad range of biochemical, molecular and cellular biology methods, we propose: 1) To determine our target panel of proteins that show evidence of specific mucin binding; 2) To determine the domains required for the mucin-protein interactions and 3) To assess the effects of mucin-protein interactions on the surface and bulk rheological properties of the mucus. If the goals of this proposal are achieved, they will increase our knowledge and understanding of the relationship between protein composition and the function of airway secretions. Such knowledge is an essential pre-requisite for informed therapeutic intervention.

描述（由申请人提供）：气道上皮粘膜屏障是维持下层上皮表面的强大先天免疫系统的核心。在最广泛的层面上，该提案的主要科学目标是阐明气道凝胶层的组织，气道凝胶层是由称为粘蛋白（MUC5B和MUC5AC）的非常大的糖蛋白通过不稳定的相互作用与其他保护性生物分子结合形成的，并评估这些相互作用在气道粘液完整性中的作用，作为治疗干预的重要先决条件。我们的假设是，正常气道粘液是高度结构化的，含有超过 100 种蛋白质，这些蛋白质形成不同的蛋白质复合物，其中许多以这些粘蛋白为中心。这些复合物构成了一个离散的分泌实体，我们称之为“粘蛋白相互作用组”。我们认为，凝胶特性是由粘蛋白和蛋白质之间的动态相互作用产生的，并且这些结构产生了粘液的特定流变特性，从而调节粘液通过咳嗽和流量清除而去除。这一提议将改变粘蛋白是唯一赋予粘液大部分流变特性的分子的范式，并最终使人们更好地理解这些相互作用作为肺部先天防御的作用。在测试这些假设时，我们使用广泛的生化、分子和细胞生物学方法，建议：1) 确定显示特定粘蛋白结合证据的目标蛋白质组； 2) 确定粘蛋白-蛋白质相互作用所需的结构域，以及 3) 评估粘蛋白-蛋白质相互作用对粘液表面和整体流变特性的影响。如果该提案的目标得以实现，将增加我们对蛋白质组成与气道分泌物功能之间关系的认识和理解。这些知识是知情治疗干预的重要先决条件。