GLYCOSYLATION OF P AERUGINOSA PILLI

铜绿菌毛的糖基化

• 批准号: 2665653
• 负责人: PETER A. CASTRIC
• 金额: $ 8.81万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-15 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2665653
• 关键词: Pseudomonas aeruginosa; SDS polyacrylamide gel electrophoresis; bacterial cytopathogenic effect; gene expression; glycosylation; mass spectrometry; nuclear magnetic resonance spectroscopy; pilus; posttranslational modifications; protein binding; protein structure function; proteoglycan; vaccine development; virulence

DESCRIPTION (Adapted from the applicant's abstract): P. aeruginosa persists as a major cause of life-threatening infections for individuals with cystic fibrosis, burns, wounds, cancer, etc. The pili of this bacterium are protein filaments which extend from the ends of the cell. These structures function as virulence factors by serving in adhesion. The long term objective of this project is to determine the role of pilus glycosylation in P. aeruginosa pathogenesis and to ascertain the importance of the glycan in pilus vaccine design. Work from Dr. Castric's laboratory has shown that the pili of strain 1244 are glycosylated. The glycan specificity in these pilin glycosylation reactions is low, a finding which has implications for the pilus vaccine design. The work proposed in this application would explore the extent of this glycan substrate specificity using both in vivo (expressing pilin genes in heterologous strains) and in vitro (glycosylation in cell free extracts) studies. He has also proposed to examine the effect of glycosylation on pilus function, such as adhesion and pilus retraction. Finally, a comprehensive structural analysis of the glycan will be performed, using mass spectrometry and NMR methodologies.

描述（改编自申请人摘要）：铜绿假单胞菌持续存在 作为一个主要的原因，危及生命的感染的个人囊性 纤维化、烧伤、创伤、癌症等。这种细菌的皮利 从细胞末端延伸的蛋白质丝。 这些结构 作为毒力因子起粘附作用。 长期 本项目的目的是确定菌毛糖基化在 P.铜绿假单胞菌的发病机制，并确定聚糖的重要性， 菌毛疫苗设计 卡斯特里博士实验室的研究表明， 菌株1244的皮利被糖基化。 这些菌毛蛋白的聚糖特异性 糖基化反应低，这一发现对 菌毛疫苗设计 本申请中提出的工作将探索 这种聚糖底物特异性的程度， （在异源菌株中表达菌毛蛋白基因）和体外（糖基化 在无细胞提取物中）研究。 他还建议研究 糖基化对菌毛功能的影响，如粘附和菌毛回缩。 最后，将对聚糖进行全面的结构分析。 使用质谱法和NMR方法进行。