YopT: A Yersinia Virulence Factor

YopT：耶尔森菌毒力因子

• 批准号: 6907659
• 负责人: JACK E DIXON
• 金额: $ 30.78万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-15 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6907659
• 关键词: X ray crystallography; Yersinia pestis; Yersinia pestis disease; antibiotics; bacteria infection mechanism; bacterial proteins; bioassay; bioterrorism /chemical warfare; chemical models; drug resistance; endopeptidases; enzyme activity; enzyme mechanism; enzyme structure; enzyme substrate complex; genetic strain; host organism interaction; protease inhibitor; protein purification; recombinant proteins; virulence

DESCRIPTION (provided by applicant): Recent acts of bioterrorism have underscored the importance of understanding the molecular mechanisms of bacterial pathogenesis. Yersinia pestis, the causative agent of the plague or the Black Death, has been used previously as a biological weapon. There is also concern that genetic engineering has been used to generate antibiotic resistant strains of Yersinia. This proposal focuses on understanding the biochemistry of the Yersinia virulence factor known as YopT. Our laboratory has recently demonstrated that YopT is a cysteine protease. We propose to develop a highly efficient and sensitive assay for YopT's catalytic activity. We will also produce and purify recombinant YopT, a YopT/SycT complex, and a catalytically inactive YopT complexed with substrate. The purified protein(s) will be used to obtain crystalline protein suitable for structural analyses using X-ray diffraction. We have chosen the R21 venue for funding because, although we have made good progress on determining the function of this virulence protein, we have no preliminary data about its catalytic properties. In addition, even though we have determined the structure of a YopT family member, AvrPphB, it will be essential to have a structure of YopT in order to identify potential inhibitors directed against this protease.

描述（由申请人提供）：最近的生物恐怖主义行为强调了了解细菌发病机制的分子机制的重要性。鼠疫耶尔森氏菌是鼠疫或黑死病的病原体，以前曾被用作生物武器。人们还担心基因工程已被用来产生耐抗生素的耶尔森氏菌菌株。该提案的重点是了解耶尔森氏菌毒力因子 YopT 的生物化学。我们的实验室最近证明 YopT 是一种半胱氨酸蛋白酶。我们建议开发一种高效且灵敏的 YopT 催化活性检测方法。我们还将生产和纯化重组 YopT、YopT/SycT 复合物以及与底物复合的无催化活性的 YopT。纯化的蛋白质将用于获得适合使用 X 射线衍射进行结构分析的晶体蛋白质。我们选择 R21 进行资助是因为，尽管我们在确定这种毒力蛋白的功能方面取得了良好进展，但我们没有关于其催化特性的初步数据。此外，尽管我们已经确定了 YopT 家族成员 AvrPphB 的结构，但为了识别针对该蛋白酶的潜在抑制剂，拥有 YopT 的结构仍然至关重要。