Pseudomonas aeruginosa 1244 Pilin Glycosylation

描述(申请人提供)：铜绿假单胞菌仍然是危及生命的感染的主要原因，有以下情况的个人：囊性纤维化、烧伤、伤口、癌症(白血病)、接受免疫抑制治疗的人、糖尿病患者以及静脉吸毒者。这种细菌的菌毛是从细胞末端延伸出来的蛋白质细丝。这些结构既是黏附因子，也是重要的毒力因子。我实验室的工作表明，铜绿假单胞菌1244的药片是与这种微生物的O抗原重复单位糖基化的。这种三糖部分起源于脂多糖O-抗原的生物合成途径。该项目的长期目标是确定菌毛糖基化在铜绿假单胞菌发病机制中的作用，并确定多糖在菌毛疫苗设计中的重要性。本提案中描述的工作旨在确定铜绿假单胞菌1244菌毛素糖基化反应中的多糖和菌毛素底物。此外，还将确定Pilo的亚细胞定位，Pilo是催化铜绿假单胞菌中毛细蛋白糖基化的酶。这种酶的膜拓扑结构也将被确定。最后，确定了铜绿假单胞菌1244菌毛蛋白糖基化反应的亚细胞定位。

DESCRIPTION (provided by applicant): Pseudomonas aeruginosa persists as a major cause of life-threatening infections for individuals with the following conditions: cystic fibrosis, burns, wounds, cancer (leukemias), those receiving immunosuppressive therapy, diabetics, as well as intravenous drug users. The pili of this bacterium are protein filaments that extend from the ends of the cell. These structures serve as adhesion factors, and so are important virulence factors. Work from my laboratory has shown that the pill of P. aeruginosa 1244 are glycosylated with the O-antigen repeating unit of this organism. This trisaccharide moiety originates in the lipopolysaccharide O-antigen biosynthetic pathway. The long-term objectives of this project are to determine the role of pilus glycosylation in P. aeruginosa pathogenesis and to ascertain the importance of the glycan in pilus vaccine design. The work described in this proposal aims to determine the glycan and the pilin substrates in the P. aeruginosa 1244 pilin glycosylation reaction. In addition, the subcellular location of PilO, the enzyme that catalyzes pilin glycosylation in P. aeruginosa will be established. The membrane topology of this enzyme will also be determined. Finally, the subcellular location of the P. aeruginosa 1244 pilin glycosylation reaction established.