INTERACTIONS OF BURKHOLDERIA NEEDLE AND TIP PROTEINS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 许多影响全世界数百万人的病原菌，包括一些被归类为生物恐怖主义潜在代理人的病原菌，在其细胞表面使用III型分泌针装置将有毒蛋白质注入人类宿主细胞。 目前还没有疫苗被批准用于对抗这些病原体中的任何一种，抗生素耐药菌株的出现使这些病原体成为对公共健康和安全的主要威胁。 由于针装置对毒力至关重要，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种有吸引力的方法。 这种方法需要详细了解针装置组装中涉及的蛋白质-蛋白质相互作用。 针由大约120个相同的蛋白质拷贝组装而成，这些拷贝以螺旋方式排列，针的尖端由几个特殊尖端蛋白质拷贝覆盖。 我们的长期目标是精确地确定针装置是如何从其蛋白质组分组装的。 在这里，我们建议阐明BsaL和BipD，类鼻疽伯克霍尔德氏菌，这是一种病原体与生物恐怖主义和地方性传染病在泰国和北方澳大利亚的针和尖端蛋白质的蛋白质-蛋白质相互作用，在细胞培养中使用NMR，诱变和细菌入侵测定的组合。 从长远来看，所获得的知识将被应用于设计小分子策略，以破坏针组装，从而使有毒细菌无毒。

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.