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.

描述（来自申请人的摘要）：假单胞菌产生几种 对于生物体在宿主中生存很重要的毒力因子 并导致角膜炎，并且对这些确定的因素加上以前的研究 未识别的因素对于理解眼部疾病的机制很重要 由假单胞菌引起的疾病。该提案的目的是 1) 确定 蛋白质转运和蛋白酶前肽结构域折叠中的功能 IV，一种被认为是角膜炎毒力因子的酶 由铜绿假单胞菌引起，2) 分离和鉴定 来自铜绿假单胞菌的未表征的蛋白酶可能对 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， 并将确定其在细菌细胞中的位置。前肽？ 还将分析成熟蛋白酶 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 将被分离并对其 N 末端进行测序，以与已发表的结果进行比较 铜绿假单胞菌的序列。