INTERACTIONS OF BURKHOLDERIA NEEDLE AND TIP PROTEINS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 许多影响全球数百万人的病原体，包括一些被归类为生物恐怖主义潜在病原体的细菌，使用其细胞表面的III型分泌针设备向人类宿主细胞注入有毒蛋白质。目前还没有针对这些病原体的疫苗被批准普遍使用，而且抗生素耐药菌株的出现使这些病原体对公众健康和安全构成了重大威胁。由于针状结构对致病力至关重要，并且暴露在细菌表面，因此破坏这种蛋白质组装是开发新型抗生素的一个有吸引力的方法。这种方法需要详细了解针状装置组装过程中涉及的蛋白质-蛋白质相互作用。针是由大约120个相同的蛋白质拷贝组成的，这些蛋白质以螺旋方式排列，针的尖端由几个特殊的尖端蛋白质组成。我们的长期目标是准确地确定针头装置是如何由其蛋白质成分组装而成的。我们建议结合核磁共振、诱变和细胞培养中的细菌入侵分析，阐明假鼻疽伯克霍尔德氏菌的针状和尖端蛋白Bsal和BipD的蛋白质-蛋白质相互作用。从长远来看，所获得的知识将被应用于设计小分子策略，以破坏针头组装，从而使毒力强的细菌变得无毒。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Many pathogenic bacteria that affect millions of people worldwide, including some that are classified as potential agents of bioterrorism, use a type III secretion needle apparatus on their cell surface to inject toxic proteins into human host cells. No vaccines are currently approved for general use against any of these pathogens, and the appearance of antibiotic resistant strains makes these pathogens major threats to public health and safety. Because the needle apparatus is critical for virulence and is exposed on the bacterial surface, disrupting this protein assembly is an attractive approach for the development of novel antibiotics. This approach requires a detailed understanding of the protein-protein interactions involved in the assembly of the needle apparatus. The needle is assembled from about 120 identical copies of a protein that are arranged in a helical manner, and the tip of needle is capped by a few copies of a special tip protein. Our long term goal is to determine precisely how the needle apparatus is assembled from its protein components. We propose here to elucidate the protein-protein interactions of BsaL and BipD, the needle and tip proteins of Burkholderia pseudomallei, which is a pathogen associated with bioterrorism and endemic infectious disease in Thailand and Northern Australia, using a combination of NMR, mutagenesis, and bacterial invasion assays in cell culture. In the longer term the knowledge gained will be applied to devising small molecule strategies to disrupt needle assembly and thereby render virulent bacteria non-virulent.