BIOSYNTHESIS OF ENTEROBACTERIAL COMMON ANTIGEN

肠杆菌共同抗原的生物合成

• 批准号: 3131290
• 负责人: PAUL D RICK
• 金额: $ 12.45万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3131290
• 关键词: Enterobacteriaceae; N acetylglucosamine; Salmonella typhimurium; bacterial antigens; bacterial polysaccharides; bacterial somatic antigen; carbohydrate biosynthesis; cell membrane; chemical chain length; chemical structure function; gel electrophoresis; gram negative bacteria; lipopolysaccharides; mutant; surface antigens; thin layer chromatography

The long-term goals of this research are directed at elucidating the mechanisms involved in the biogenesis and assembly of the outer membrane of gram-negative bacteria. It is anticipated that a knowledge of this process will provide a rational basis for the development of new antimicrobial agents and also provide insights into the processes involved in membrane biogenesis and assembly in eukaryotic cells and cell organelles. The specific focus of this research is to define the mechanism of biosynthesis and assembly of enterobacterial common antigen (ECA). ECA is an outer membrane glycolipid that is unique to the Enterobacteriaceae, and it occurs in all bacteria belonging to this family. We have investigated ECA synthesis in Salmonella tymphimurium and Escherichia coli, and both in vivo and in vitro systems have been successfully developed and employed for this purpose. We wish to continue our studies, and the specific aims for the requested period of support are (a) to demonstrate that Fuc4NAc- ManNAcA-GlcNAc-pyrophosphorylundecaprenol is a precursor of ECA heteropolysaccharide chains; (b) to determine the mechanism involved in' the assembly of ECA chains; (c) to determine the mechanism involved in the synthesis of phosphoglyceride-linked ECA chains (ECApG); (d) to determine the mechanism involved in the synthesis of lipopolysaccharide-linked ECA (ECALPS); and (3) to isolate and structurally characterize polyisoprenoid-linked intermediates involved in ECA synthesis. The proposed research will utilize techniques of molecular biology and biochemistry including in vivo pulse-labeling, enzymology, and the isolation and structural characterization of biosynthetic intermediates.

这项研究的长期目标旨在阐明 参与外膜生物发生和组装的机制 革兰氏阴性细菌。预计这方面的知识 过程将为新产品的开发提供合理的基础 抗菌剂，并提供对过程的深入了解 参与真核细胞膜的生物合成和组装 细胞器。 本研究的具体重点是明确其机制 肠细菌共同抗原（ECA）的生物合成和组装。出口管制局 是一种独特的外膜糖脂 肠杆菌科，它存在于属于该科的所有细菌中 家庭。我们研究了鼠伤寒沙门氏菌中 ECA 的合成 和大肠杆菌，体内和体外系统均已 为此目的已成功开发并采用。我们希望 继续我们的学习，以及所要求的时期的具体目标 支持是 (a) 证明 Fuc4NAc- ManNAcA-GlcNAc-焦磷酸十一异丙烯醇是 ECA 的前体 杂多糖链； (b) 确定涉及的机制' ECA链条的组装； (c) 确定涉及的机制 磷酸甘油酯连接的 ECA 链 (ECApG) 的合成； (d) 至 确定参与合成的机制 脂多糖连接的 ECA (ECALPS)； (3) 隔离和 结构表征涉及的聚异戊二烯连接的中间体 在ECA合成中。 拟议的研究将利用分子生物学技术和 生物化学，包括体内脉冲标记、酶学和 生物合成中间体的分离和结构表征。