BORDETELLA CYCLASE--STRUCTURE AND BIOLOGICAL ACTIVITIES

产品描述：腺苷酸环化酶（AC）毒素是一种新的蛋白质细菌 百日咳杆菌毒素百日咳杆菌产生的毒素，是百日咳的病原体 咳嗽. 这种毒素的序列是已知的，并且单一的 赖氨酸残基已被证明是必不可少的毒素活动。 然而，关于毒素介导的细胞凋亡的机制知之甚少。 过程或其功能相互关系。 该提案将集中于AC毒素所涉及的各个步骤 行动上将调整或开发检测方法，以实现分离和 表征途径中的每个单独步骤。方法论 方法包括使用固有的色氨酸荧光 荧光能量传递、脂质双层膜和全细胞膜片钳 电导、电子显微镜和X射线衍射。功能 研究将使用随机和特定地点的 诱变以产生毒素缺陷的AC毒素突变体 和/或溶血素活性。 类似地，体外酰化将 提供了一种用棕榈酰以外的物质制备毒素的机制， LYs983上的基团，以确定酰化的作用。额外 将用CD光谱和X射线衍射收集结构数据 研究，以更好地了解AC毒素的作用。

DESCRIPTION: Adenylate cyclase (AC) toxin is a novel protein bacterial toxin produced by Bordetalla pertussis, the causative agent of whooping cough. The sequence of this toxin is known and palimiolyation of a single Lys residue has been demonstrated to be essential for toxin activities. Nevertheless, little is known about mechanisms for toxin-mediated processes or their functional interrelationships. This proposal will focus on the individual steps involved in AC toxin action. Assays will be adapted or developed to enable the isolation and characterization of each individual step in the pathway. The methodologic approaches will include use of intrinsic tryptophan fluorescence fluoresecence energy transfer, lipid bilayer and whole cell patch-clamp conductance, electron microscopy and x-ray diffraction. The functional studies will be complemented with th use of random and site-specific mutagenesis to generate mutants of AC toxin that are defective in toxin and/or hemolysin activities. Similarly, an in vitro acylation will provide a mechanism for preparation of toxins with other than palmitoyl groups on LYs983, in order to determine the role of acylation. Additional structural data will be collected with CD spectroscopy an x-ray diffraction studies to develop a better understanding of the actions of AC toxin.