STRUCTURE DETERMINATION OF PERTUSSIS TOXIN

百日咳毒素的结构测定

• 批准号: 3145790
• 负责人: CLARENCE E SCHUTT
• 金额: $ 13.92万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-03-01 至 1996-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3145790
• 关键词: ADP ribosylation; X ray crystallography; biological signal transduction; enzyme mechanism; enzyme structure; epitope mapping; immunogenetics; immunoregulation; mutant; nucleoside ribosyltransferase; pertussis toxin; protein structure function; protein transport; site directed mutagenesis

It is proposed to determine the three-dimensional structure of pertussis toxin by high resolution x-ray crystallography. Pt is a protein exotoxin produced by Bordetella pertussis, the bacterium that causes the infectious disease known as whooping cough, a disease affecting over 60 million infants and leading to over one million deaths per year world-wide. A specific objective of this research is to define those structural features of PT that are important in eliciting the protective immune response after natural infection or vaccination. Although protective vaccines prepared rom unfractionated B. pertussis cells have long been available, they are often associated with adverse side effects including rare cases of neurological impairment. The development of improved vaccines, free of deleterious complications, requires the definition of the minimal molecular structures against which the protective response is mobilized. Since pertussis toxin is considered a major virulence factor in the disease, the specific immune response to PT is almost certainly essential for protection. A high resolution structure of Pt in conjunction with available genetic and biochemical data would allow recognition of structural features involved in the immunogenicity, enabling a distinction to be made between those regions of the molecule conferring protective immunity from those simply eliciting a response. Another important reason for obtaining more detailed structural information concerning PT is that its mode of action is the interference with a critical component of the fundamental signal transduction machinery of eukaryotic cells. The current unifying hypothesis is that a wide variety of receptor mediated events at the membranes are effected by a class of proteins known as the G proteins. It is known that Pt catalyses the covalent modification of G proteins by transferring ADP-ribose from NAD+. Finally, an important aim of the work is to better understand the basic principles involved in mechanisms by which proteins such as the A subunit of PT are internalized by cells at the membrane after interaction with translocating complexes like the B subunit of PT. A three-dimensional structure of PT could lead to more effective and safest vaccines against whooping cough, provide a basis for designing cytotoxic agents coupled to cancer cells targeting antibodies, and contribute to our improved understanding of basic receptor biology and oncogenesis.

提出确定百日咳的三维结构 通过高分辨率 X 射线晶体学分析毒素。 Pt是一种蛋白质外毒素 由百日咳博德特氏菌（Bordetella pertussis）产生，这种细菌引起传染性 百日咳是一种影响超过 6000 万人的疾病 婴儿，每年导致全世界超过一百万人死亡。 一个 这项研究的具体目标是定义这些结构特征 PT 对引发保护性免疫反应很重要 自然感染或疫苗接种。尽管已准备好保护性疫苗 未经分级的百日咳博德特氏菌细胞早已可用，它们通常 与不良副作用相关，包括罕见的神经系统疾病病例 损害。 开发不含有害物质的改良疫苗 复杂化，需要定义最小的分子结构 动员保护性反应。 由于百日咳毒素 被认为是该疾病的主要毒力因素，特异性免疫 对 PT 的反应几乎肯定对于保护至关重要。 一个高 Pt 的解析结构与可用的遗传和 生化数据将允许识别涉及的结构特征 免疫原性，能够区分这些区域 赋予保护性免疫力的分子，使其免受那些简单引发的 一个回应。 获得更详细信息的另一个重要原因 关于PT的结构信息是它的作用方式是 干扰基本信号的关键成分 真核细胞的转导机制。 目前统一 假设是多种受体介导的事件 细胞膜受到一类称为 G 蛋白的蛋白质的影响。 它 众所周知，Pt 通过以下方式催化 G 蛋白的共价修饰： 从 NAD+ 转移 ADP-核糖。 最后，工作的一个重要目标 是为了更好地理解机制所涉及的基本原理 PT 的 A 亚基等蛋白质被细胞内在化 与 B 亚基等易位复合物相互作用后的膜 PT。 PT 的三维结构可以带来更有效的结果 和最安全的百日咳疫苗，为设计提供基础 与靶向抗体的癌细胞偶联的细胞毒性剂，以及 有助于我们提高对基本受体生物学的理解 肿瘤发生。