Pseudomonas proteases as ocular virulence factors

描述(来自申请人？S摘要)：假单胞菌产生几种 对生物体在宿主体内生存很重要的毒力因子 并导致角膜炎，以及对这些已确定的因素的研究 未知因素对于了解眼球运动的机制很重要。 由假单胞菌引起的疾病。这项提案的目的是1)确定 蛋白水解酶前肽段在蛋白质转运和折叠中的作用 IV，一种被认为是角膜炎致病因子的酶 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知蛋白酶，可能是重要的 了解这种形式的角膜炎。蛋白酶IV的前肽结构域将是 克隆、表达和纯化以制造抗体的目的， 它在细菌细胞中的位置将被确定。前肽？S 此外，还将分析成熟蛋白酶IV分泌过程中的可能功能。一个 假单胞菌提取液酶谱上观察到的未知蛋白酶 将被分离出来，并对其N末端进行测序，以便与已发表的 铜绿假单胞菌的序列。

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.