Pseudomonas proteases as ocular virulence factors

假单胞菌蛋白酶作为眼部毒力因子

• 批准号: 6404863
• 负责人: MARY E MARQUART
• 金额: $ 4.2万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6404863
• 关键词: Pseudomonas aeruginosa; bacterial proteins; endopeptidases; keratitis; postdoctoral investigator; protein folding; protein localization; protein purification; protein structure function; protein transport; virulence

DESCRIPTION (From the Applicant?s Abstract): Pseudomonas produces several virulence factors that are important for the organism to survive in the host and cause keratitis, and the study of these identified factors plus previously unidentified factors is important for understanding the mechanisms of ocular disease caused by Pseudomonas. The aims of this proposal are 1)to determine the functions in protein transport and folding of the propeptide domain of protease IV, an enzyme that has been implicated as a virulence factor in keratitis caused by Pseudomonas aeruginosa, and 2)to isolate and identify an uncharacterized protease from P. aeruginosa that could be important for understanding this form of keratitis. The protease IV propeptide domain will be cloned, expressed, and purified for the purpose of making an antibody to it, and its location in the bacterial cell will be determined. The propeptide?s possible function in mature protease IV secretion will also be analyzed. An unknown protease that has been observed on zymograms of Pseudomonas extracts will be isolated and its N-terminus sequenced for comparison to the published sequence of P. aeruginosa.

申请人（申请人）？s摘要）：假单胞菌产生几个 对生物体在宿主中生存至关重要的毒力因子 并导致角膜炎，对这些确定的因素的研究加上以前的研究， 不明因素对于理解眼内炎的机制是重要的， 由假单胞菌引起的疾病。本提案的目的是：（1）确定 蛋白酶前肽结构域在蛋白质转运和折叠中的功能 IV，一种被认为是角膜炎毒力因子的酶 引起的铜绿假单胞菌，和2）分离和鉴定 来自铜绿假单胞菌的未表征的蛋白酶，其可能对 了解这种形式的角膜炎。蛋白酶IV前肽结构域将是 克隆、表达和纯化以制备其抗体， 并确定其在细菌细胞中的位置。前肽？S 还将分析在成熟蛋白酶IV分泌中的可能功能。一个 在假单胞菌提取物的酶谱上观察到的未知蛋白酶 将其分离并对其N-末端测序，以与已发表的 铜绿假单胞菌的序列。