Structure and action of CARDS toxin

卡毒素的结构和作用

基本信息

批准号：
8182002
负责人：
Peter JOHN HART
金额：
$ 31.66万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8182002
关键词：
3-Dimensional ADP Ribose Transferases Acute Allergic inflammation Amino Acid Sequence Amino Acids Antibiotic Therapy Asthma Biochemical Blood Vessels Bordetella pertussis C-terminal Catalytic Domain Cell Surface Receptors Chest Chronic Clinical Collaborations Community Acquired Respiratory Distress Syndrome Toxin Complex Diagnostic Disease Etiology Exotoxins Exposure to Extracellular Domain Fab Immunoglobulins Goals Human Inflammation Inflammatory Lung Lung diseases Lymphocyte Mediating Molecular Structure Mus Mycoplasma pneumonia infection Mycoplasma pneumoniae Papio Pathogenesis Pathology Patients Peptide Sequence Determination Pertussis Play Pneumonia Progress Reports Proteins Pulmonary Surfactant-Associated Protein A Recombinants Reporting Research Resolution Respiratory System Respiratory tract structure Role Severities Structure Symptoms Techniques Testing Therapeutic Agents Time Toxin Virulence Virulence Factors Work X ray diffraction analysis X-Ray Diffraction airway obstruction airway remodeling cofactor design inhibitor/antagonist neutralizing monoclonal antibodies novel diagnostics novel therapeutics pathogen pulmonary function receptor binding research study respiratory small molecule three dimensional structure tool

项目摘要

Asthma is a complex disorder characterized by episodic airway obstruction and hyper-responsiveness, sometimes accompanied by airway remodeling. Although the underlying causes of asthma remain poorly understood, one contributing factor is exposure to respiratory pathogens. For example, asthmatics positive for Mycoplasma pneumoniae (M. pneumoniae) infection have demonstrated an improvement in pulmonary function after antibiotic treatment whereas patients that test negative do not, suggesting a causal relationship between M. pneumoniae infection and asthma symptom severity. Although there is a strong clinical correlation between M. pneumoniae infection and a sub-set of asthma cases, until recently, the identification of a virulence factor that might play a role in disease pathogenesis had remained elusive. This situation changed, however, when we discovered a 591 amino acid protein with ADP-ribosyltransferase (ART) activity in M. pneumoniae designated Community Acquired Respiratory Distress Syndrome ToXin (CARDS TX). The experiments outlined in this proposal are designed to uncover the structure and action of CARDS TX using a range of biophysical techniques. Using the well-established tools of single crystal X-ray diffraction, we will determine: 1) the three-dimensional structure of CARDS TX; 2) CARDS TX in complex with its NAD"" co-factor; 3) CARDS TX in complex with neutralizing monoclonal antibody Fab fragments; and 4) CARDS TX in complex with recombinant extracellular domains of its cell surface receptor surfactant protein-A (SP-A). A longer term goal is to use these 3-D structures in conjunction with the information coming from Projects 1 and 2 as platforms for the design of CARDS TX inhibitors, which may represent novel therapeutic agents for the treatment of asthma and pulmonary inflammation.

哮喘是一种复杂的疾病，其特征是间歇性气道阻塞和高反应性，有时并伴有气道重塑。尽管哮喘的根本原因仍知之甚少，但其中之一影响因素是接触呼吸道病原体。例如，支原体阳性的哮喘患者肺炎支原体（肺炎支原体）感染在抗生素治疗后显示肺功能有所改善治疗，而检测呈阴性的患者则不接受治疗，这表明肺炎支原体之间存在因果关系感染和哮喘症状的严重程度。尽管肺炎支原体与肺炎支原体之间存在很强的临床相关性。感染和哮喘病例的一个子集，直到最近，才确定了可能发挥作用的毒力因子疾病的发病机制仍然难以捉摸。然而，当我们发现 591 个氨基酸时，这种情况发生了变化。社区获得性肺炎支原体中具有 ADP-核糖基转移酶 (ART) 活性的酸性蛋白呼吸窘迫综合征毒素 (CARDS TX)。本提案中概述的实验旨在使用一系列生物物理技术揭示 CARDS TX 的结构和作用。使用成熟的利用单晶X射线衍射工具，我们将确定：1）CARDS TX的三维结构； 2） CARDS TX 与其 NAD"" 辅因子复合； 3) CARDS TX 与中和单克隆抗体 Fab 复合碎片； 4) CARDS TX与其细胞表面受体的重组胞外结构域形成复合物表面活性剂蛋白-A (SP-A)。长期目标是将这些 3D 结构与信息结合使用来自项目 1 和 2 作为 CARDS TX 抑制剂设计的平台，这可能代表新颖用于治疗哮喘和肺部炎症的治疗剂。