Structural Interactions of Bacterial Adhesin with Glycosaminoglycans

抗生素耐药性正在成为迫在眉睫的健康威胁。对抗这种情况的一种策略是抑制 微生物与人体细胞的粘连。然而，微生物与靶标之间的特定结构相互作用 细胞特性不好，从而阻碍了新的抗微生物药物的开发。这 Proposal致力于通过对相互作用进行深入的结构性研究来克服这一缺陷 在细菌粘附素和糖胺聚糖(GAG)之间，是一种普遍存在的多糖，存在于 细胞表面，细菌粘附素的主要靶点。提案的重点将放在装饰上 结合蛋白(DBP)，一种来自伯氏疏螺旋体的粘附素，是莱姆病的病原体。学习 GAGS与DBP的相互作用及DBP在胞浆及其天然脂蛋白中的结构 表格将使用溶液核磁共振解决。以这些结构为基础，DBP与 将确定多相和均相的GAG低聚物。这项研究的主要目的是 获得与GAG络合的DBP的高分辨结构，并从这些结构中推断出特定的 GAG和DBP之间的相互作用，可以靶向防止细菌与人类细胞的黏附。 最后，由于GAG通常以蛋白多糖的形式存在，因此将尝试表征 DBP和完整的核心蛋白聚糖之间的相互作用，这是DBP主要针对的蛋白多糖。的主要目标是 提案的这一部分将构建DBP的复杂结构，仅使用完整的装饰素 核磁共振衍生的取向限制和完整装饰素的二聚性。 拟议的工作将在复杂碳水化合物研究中心进行，这是一个多学科的 该研究所是全国首屈一指的糖生物学研究中心之一。候选人的长期任期 职业目标是了解GAG与其蛋白质靶标相互作用的规则，并设计 将这些互动引向所需方向的策略。候选人在竞选期间的直接目标 指导阶段将熟练掌握用于净化的化学和生物化学方法 GAG寡聚体，并学习哺乳动物表达系统的使用，这对获得完整的 装饰品。对这些技术的了解将使应聘者能够在 建议的地区。

Antibiotic resistance is becoming an imminent health threat. One strategy for combating this is to inhibit the adhesion of microbes to human cells. However, specific structural interactions between microbes and target cells are not well characterized, thus hampering the development of new anti-microbial agents. This proposal strives to overcome this deficiency by conducting an in-depth structural study of the interactions between bacterial adhesins and glycosaminoglycans (GAGs), a ubiquitous polysaccharide found on the surface of cells and a primary target for bacterial adhesins. The focus of the proposal will be on the decorin binding protein (dbp), an adhesin from Borrelia burgdorferi, the etiological agent of Lyme disease. To study the interactions between GAGs and dbp, the structures of dbp in both cytoplasmic and its native lipoprotein form will be solved using solution NMR. Using these structures as bases, dbp's interactions with heterogeneous and homogeneous GAG oligomers will be determined. The main goal of this research is to obtain high resolution structures of dbp complexed to GAG and deduce from these structures the specific interactions between GAG and dbp that can be targeted to prevent the adhesion of bacteria to human cells. Finally, as GAGs often exist in the form of proteoglycans, an attempt will be made to characterize the interactions between dbp and intact decorin, the proteoglycan that dbp primarily targets. The main goal for this portion of the proposal will be to construct the complex structure of dbp with intact decorin utilizing only NMR derived orientational restraints and the dimeric nature of intact decorin. The proposed work will be carried out at the Complex Carbohydrate Research Center, a multidisciplinary research institute and one of the country's premier glycobiology research centers. The candidate's long term career goal is to understand the rules governing the interactions of GAGs with its protein targets and devise strategies for steering these interactions in a desired direction. The candidate's immediate goal during the mentored phase will be to acquire proficiency with the chemical and biochemical methods used to purify GAG oligomers and learn the use of mammalian expression systems that will be crucial for obtaining intact decorin. The knowledge of these techniques will enable the candidate to conduct independent research in the proposed area.